Beneficial effect of garlic on d-galactosamine and lipopolysaccharide-induced acute hepatic failure in male albino rats

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Cirera, L. García-Marcos, J. Giménez, S. Moreno-Grau, A. Tobías, V. Pérez-Fernández, B. Elvira-Rendeles, J.J. Guillén, C. Navarro" "autores" => array:9 [ 0 => array:2 [ "nombre" => "L." "apellidos" => "Cirera" ] 1 => array:2 [ "nombre" => "L." "apellidos" => "García-Marcos" ] 2 => array:2 [ "nombre" => "J." "apellidos" => "Giménez" ] 3 => array:2 [ "nombre" => "S." "apellidos" => "Moreno-Grau" ] 4 => array:2 [ "nombre" => "A." "apellidos" => "Tobías" ] 5 => array:2 [ "nombre" => "V." "apellidos" => "Pérez-Fernández" ] 6 => array:2 [ "nombre" => "B." "apellidos" => "Elvira-Rendeles" ] 7 => array:2 [ "nombre" => "J.J." "apellidos" => "Guillén" ] 8 => array:2 [ "nombre" => "C." "apellidos" => "Navarro" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054611002497?idApp=UINPBA00004N" "url" => "/03010546/0000004000000004/v1_201304101110/S0301054611002497/v1_201304101110/en/main.assets" ] "en" => array:19 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Beneficial effect of garlic on d-galactosamine and lipopolysaccharide-induced acute hepatic failure in male albino rats" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "238" "paginaFinal" => "243" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Bahaa K.A. Abdel-Salam, Abd-Alla A.A. Sayed" "autores" => array:2 [ 0 => array:4 [ "nombre" => "Bahaa K.A." "apellidos" => "Abdel-Salam" "email" => array:1 [ 0 => "Manar_Muhamad@yahoo.com" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "¿" "identificador" => "cor0005" ] ] ] 1 => array:2 [ "nombre" => "Abd-Alla A.A." "apellidos" => "Sayed" ] ] "afiliaciones" => array:1 [ 0 => array:1 [ "entidad" => "Zoology Department, Faculty of Science, Minia University, El-Minia 61519, Egypt" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1230 "Ancho" => 2090 "Tamanyo" => 139052 ] ] "descripcion" => array:1 [ "en" => "Changes in IL-4 levels in d-GalN/LPS injected groups versus control group and garlic treated groups. Significant differences between all groups, P<0.05, were monitored." ] ] ] "textoCompleto" => "IntroductionLipopolysaccharide (LPS), a component of the cell wall of Gram-negative bacteria, is generally considered to be a major pathogenic element in Gram-negative bacterial infection, and can eventually cause systemic inflammatory response syndrome.<a class="elsevierStyleCrossRef" href="#bib0005">1</a>The innate immune system copes with infection by producing pro-inflammatory mediators such as tumour necrosis factor-α (TNF-α); however, overproduction of these mediators eventually leads to systemic inflammatory response syndrome.<a class="elsevierStyleCrossRef" href="#bib0010">2</a> LPS stimulates monocytes/macrophages via Toll-like receptor 4 (TLR4), which belongs to the TLR family.<a class="elsevierStyleCrossRef" href="#bib0015">3</a> LPS is recognised by TLR4 together with accessory molecules such as myeloid differentiation protein-2 and CD14, and TLR4 then transduces LPS signalling via both myeloid differentiation factor 88 (MyD88)-dependent and MyD88-independent pathways, each of which activates nuclear factor-κB (NF-κB).<a class="elsevierStyleCrossRef" href="#bib0020">4</a>d-Galactosamine (d-GalN) increases the susceptibility of mice to LPS-induced shock by impairing liver metabolism.<a class="elsevierStyleCrossRef" href="#bib0025">5</a> In contrast to high dose LPS-induced shock which induces a systemic disorder including multiple organ failures,<a class="elsevierStyleCrossRef" href="#bib0030">6</a> liver is a major target organ after challenge with low doses of LPS in conjunction with d-GalN.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> Similarly to high dose LPS-induced shock, TNF-α plays a central role in low dose LPS-induced shock/liver injury.<a class="elsevierStyleCrossRef" href="#bib0040">8</a>Endotoxin makes macrophages, neutrophils, and other inflammatory cells release cytokines, such as TNF-α, in great amounts, which may render inflammatory reactions out of control, ultimately resulting in sepsis, septic shock, or multiple organ failure syndromes.<a class="elsevierStyleCrossRefs" href="#bib0010">2,9</a> The release of TNF-α is the hallmark of the cellular response to the activation of the innate immune system.<a class="elsevierStyleCrossRef" href="#bib0050">10</a> The essential role of TNF in control of several infections was documented.<a class="elsevierStyleCrossRef" href="#bib0055">11</a>A critical degree of liver cell death due to apoptosis or necrosis is fundamental to the development of fulminant hepatic failure.<a class="elsevierStyleCrossRef" href="#bib0060">12</a> Activation of the pro-inflammatory cytokine cascade, including TNF-alpha is considered to play an important role in the pathophysiology and clinical outcome of this severe liver injury.<a class="elsevierStyleCrossRef" href="#bib0065">13</a> Kupffer cells, resident macrophages of the liver, have a transmembrane protein TLR-4, which recognises LPS and mediates macrophage activation and pro-inflammatory cytokine release.<a class="elsevierStyleCrossRefs" href="#bib0070">14,15</a> Upregulated TLR4 expression and function was observed in various situations of experimental liver injury.<a class="elsevierStyleCrossRefs" href="#bib0075">15–18</a> Liver macrophages are reported to be increased in number and considered to be closely related to the upregulation of cytokines and chemokines in fulminant hepatic failure.<a class="elsevierStyleCrossRef" href="#bib0095">19</a>Garlic is a sulphur-rich phytomedicine with a long history of therapeutic use. It has been shown to be useful as an antiplatelet,<a class="elsevierStyleCrossRef" href="#bib0100">20</a> antihypertensive,<a class="elsevierStyleCrossRef" href="#bib0105">21</a> antifungal,<a class="elsevierStyleCrossRef" href="#bib0110">22</a> anticancer,<a class="elsevierStyleCrossRef" href="#bib0115">23</a> antioxidant,<a class="elsevierStyleCrossRef" href="#bib0120">24</a> hypolipidaemic<a class="elsevierStyleCrossRef" href="#bib0125">25</a> and hypoglycaemic.<a class="elsevierStyleCrossRef" href="#bib0130">26</a> Its variety of therapeutic actions probably relates to the complex nature of the chemistry of garlic. It contains many sulphur-rich derivatives of the amino acid cysteinedalliin. The immunomodulatory effects of garlic have been demonstrated on a small variety of cells and tissues. The effects of garlic on human and murine peripheral blood mononuclear cells (PMBCs) and neutrophils have been investigated. Although not investigating the effect of garlic on cytokine production. Therefore, the aim of the present study is to examine the in vivo effect of garlic on normal and d-GalN/LPS treated liver tissues. We directed our efforts at the effect of garlic on the inflammatory cytokine balance of the liver tissue. We wished to examine garlic's potential to increase anti-inflammatory, and decrease inflammatory mediator production, and thus explore its use as a substance with the potential to enhance or correct liver functions.Materials and methodsReagentsd-GalN and LPS were obtained from Sigma (St. Louis, MO, USA). ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). All other chemicals were obtained commercially as reagent-grade products.Experimental animalsMale albino rats, Rattus norvegicus (6–8 weeks old, weight 100–120g) were purchased from the Biological Supply Center, Theodore Bilharz Research Institute, TBRI, Cairo, Egypt and housed under specific pathogen-free conditions and maintained on a 12-h light–dark cycle, with food and water ad libitum. Animals were classified into 12 groups (ten animals each). After rats were fasted for 18h, the first group (negative control) was untreated; the second group (first positive control) was injected with the same amount of saline intraperitoneally; the third group (second positive control) was allowed free to feed on a crude garlic used as a food supplement (5g) daily for one week before d-GalN/LPS injection. The groups from four to eleven were injected intraperitoneally with d-GalN at 500mg/kg and LPS at 50μg/kg (dissolved in 500ml of saline) to induce liver injury; the fifth, seventh, ninth and twelfth groups were allowed free to feed on garlic daily for one week after d-GalN/LPS injection. Blood samples were collected from tail at 6, 12, and 24h after d-GalN/LPS injection and at one week before and one week after d-GalN/LPS injection.Cytokines in serumSeventy-two hours after the last treating, 10 animals from each group were sacrificed under chloroform anaesthesia. Blood samples were taken and centrifuged at 3000rpm for 30min. Sera were removed and kept at −20°C for the estimation of TNF-α and IL-4 levels.Leukocytes differential countFreshly collected blood samples of about 20μl were spread on clean slides into a thin film using another smooth-edged glass slide. Each blood smear was left to air dry before being fixed with methanol for 2–3min and then labelled by the number of the animal. Blood smears were stained with 10% Giemsa's stain (Aldrich) in buffered distilled water containing 0.021M Na2HPO4/0.015M KH2PO4. pH 7–7.2 for 30min far from sun light. After that, the stain was removed by gentle washing with distilled water and the slides were air-dried at room temperature. Using light microscopy at 400× magnification, different types of blood leukocytes were recorded. At least double smears for each blood samples were counted.Histopathological examinationLiver samples were dissected after DGalN/LPS injection and garlic treating, fixed in 10% formal saline, embedded in paraffin, and cut into 5μm thickness in microtome. These sections were collected in slides and then stained with haematoxylin and eosin. The stained sections were examined under low and high power by using an Olympus microscope.Statistical analysisData were analysed using SPSS program version 13.0. Statistical analysis of the obtained data was performed using one way analysis of variance (ANOVA) test followed by least square differences (LSD) analysis for comparison between means. Results were expressed as mean±standard error (SE). Values of P<0.05 were considered statistically significant, while value of P>0.05 were considered statistically non-significant.ResultsEstimation of TNF-αSerum TNF-α levels were demonstrated in <a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>, where the negative control group (1405.26±5.252pg/ml), the first PBS positive control (1431.59±10.692pg/ml) and the second positive control pre-treated with garlic one week before d-GalN/LPS injection (1361.48±8.302pg/ml) is higher than the d-GalN/LPS treated group after 6h (172.16±2.049pg/ml), 12h (880.87±1.717pg/ml) and 24h (969.73±3.114pg/ml) which is lower than the d-GalN/LPS group treated with garlic (1129.73±3.650pg/ml, 1726.02±4.376pg/ml and 1561.90±6.574pg/ml at 6, 12 and 24h, respectively). In contrast, the garlic treated group after one week of d-GalN/LPS injection (1466.51±1.451pg/ml) is higher than the d-GalN/LPS injected groups at 6, 12 and 24h (172.16±2.049pg/ml, 880.87±1.717pg/ml and 969.73±3.114pg/ml, respectively). These results showed a significant difference in TNF-α level between all groups (P<0.05).<elsevierMultimedia ident="fig0005"></elsevierMultimedia>Estimation of IL-4Results of ELISA in <a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a> showed that IL-4 recorded 667.72±7.270pg/ml in negative control group (untreated), 640.09±3.176pg/ml in the first PBS positive control and 687.00±4.078pg/ml in the second positive control. Liver injury group injected with d-GalN/LPS counted low levels of IL-4 at 6h (283.50±1.417pg/ml), 12h (368.10±3.607pg/ml) and 24h (454.20±1.994pg/ml) than the d-GalN/LPS injected groups treated with garlic (555.75±1.864pg/ml, 813.59±2.081pg/ml and 769.63±3.421pg/ml, at the same time, respectively). In addition, the d-GalN/LPS injected groups at 6, 12 and 24h are lower than the garlic treated group after one week of d-GalN/LPS injection (751.01±1.789pg/ml). Statistical analysis showed that there is a significant difference in IL-4 between the liver injury groups and either the control group or the d-GalN/LPS injected groups treated with garlic, P<0.05.<elsevierMultimedia ident="fig0010"></elsevierMultimedia>Leukocyte differential count<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a> shows that granulocytes percentage in the garlic treated groups before (57.60±0.509pg/ml) and after (57.60±0.509pg/ml) the d-GalN/LPS injection, the negative control group (57.20±0.374pg/ml), the first positive control group treated saline (57.20±0.374pg/ml) are nearly similar in population and higher than the other groups ranged from 53.80±0.374pg/ml to 55.80±0.583pg/ml, except for the d-GalN/LPS injected group after 12h which showed the highest population (63.80±0.663pg/ml).<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>In contrast to granulocytes, the lymphocytes and monocytes percentages showed the lowest population in the d-GalN/LPS injected group after 12h (30.20±0.663pg/ml and 6.00±0.316pg/ml, respectively), <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>.Statistical analysis showed that there is a significant difference in leukocytes counts between the liver injury groups and either the control group or the groups treated with garlic, P<0.05.Histopathological findingsHistopathological examination of liver sections from control animals revealed normal architecture, <a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>A. d-GalN/LPS-intoxicated rat liver section showed loss of architecture and prominent inflammatory collections in the central rather than around central vein, <a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>B. In contrast, a slight inflammatory reaction was found in liver section of rat pre-treated with garlic prior to d-GalN/LPS challenge, <a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>C.<elsevierMultimedia ident="fig0015"></elsevierMultimedia>DiscussionMany approaches to suppress the effects of inflammatory mediators have been unsatisfactory for the effective treatment of Gram-negative bacterial.<a class="elsevierStyleCrossRef" href="#bib0010">2</a> This promoted us to use a medicinal plant, garlic, to treat bacterial infection causing liver injury.This paper describes participation of d-GalN and LPS in liver injury and points to garlic as a critical medicinal plant of resistance to this injury. Histological disorder is accompanied with the level of some cytokines, such as the pro-inflammatory TNF-α and the anti-inflammatory IL-4, and leukocytes proliferation.The liver is an important organ within the body with a central role in metabolic homeostasis, as it is responsible for the metabolism, synthesis, storage, and redistribution of nutrients, carbohydrates, fats, and vitamins. Hepatitis is the best-known liver disease.Acute liver injury was induced by d-GalN/LPS.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> The advantage of this induction is that d-GalN can potentiate the toxic effects of LPS and produce fulminant hepatitis within a few hours.<a class="elsevierStyleCrossRef" href="#bib0135">27</a> In the present study, we demonstrated that pre-treatment with garlic attenuated liver injury caused d-GalN/LPS, where the administration of d-GalN/LPS significantly down-regulated the expression of both serum TNF-alpha and IL-4 that are up-regulated with garlic.NF-kappa B plays a key role in the regulation of TNF-alpha transcription stimulated by LPS.<a class="elsevierStyleCrossRef" href="#bib0140">28</a> In the latent state, NF-kappa B is found in cytoplasm. After exposure to LPS, NF-kappa B is translocated to the nucleus and binds to NF-kappa B promoter sites on DNA, activating gene transcription of cytokines such as TNF-alpha.<a class="elsevierStyleCrossRef" href="#bib0145">29</a>Taken together, our results revealed that the up-regulation of TNF-alpha of garlic against d-GalN/LPS-induced liver injury in rat could be related to inhibiting the activation of NF-kappa B. In addition, the down-regulation of the TNF-alpha might also be due to the death of the Kupffer cells, resident macrophages of the liver, where they have a transmembrane protein Toll-like receptor 4 (TLR4), which recognises endotoxin (LPS) and mediates macrophage activation and pro-inflammatory cytokine release.<a class="elsevierStyleCrossRef" href="#bib0070">14</a>The histological injuries such as inflammation induced by d-GalN/LPS were markedly improved by garlic, which showed a protective effect as a control. Previous studies showed that LPS exerted the toxic effects mainly through the generation of endogenous inflammatory cytokines secreted from macrophages. Among these cytokines, TNF-α is a key factor that contributes to the triggering of an inflammatory cascade involving the induction of cytokines including interferon-γ, nitric oxide, IL-1β and cell adhesion molecules, etc.<a class="elsevierStyleCrossRef" href="#bib0150">30</a>A proper balance between pro- and anti-inflammatory mediators is necessary for modulating an adequate immune response toward LPS. IL-4 is known to be an anti-inflammatory cytokine which can suppress serum TNF-α levels and reduce the mortality of mice exposed to d-GalN/LPS.<a class="elsevierStyleCrossRef" href="#bib0155">31</a> In the present study, there was a massive induction of the inflammatory cytokines TNF-α in hepatic failure caused by d-GalN/LPS, which were not counterbalanced by the anti-inflammatory cytokine IL-4. This imbalance may promote inflammatory responses to liver damage. After administration of garlic, serum IL-4 maintained at a higher level, which indicated that the imbalance was reversed by garlic to a certain extent.Interaction between T-cells and hepatocytes that was regulated by adhesion molecules is dysregulated in harmful inflammatory processes. During the d-GalN/LPS intoxication, the expressions of ICAM-1 and LFA-1 in hepatocytes and non-parenchymal cells were enhanced. The elevation of these molecules was essential to trigger the extravasations of neutrophils into the liver parenchyma, which produced cytotoxic damage to hepatocytes.<a class="elsevierStyleCrossRef" href="#bib0160">32</a> This is in accordance with the high percentage of neutrophils monitored in our results. Previous studies suggested that TNF-α could induce the transcription of adhesion molecules.<a class="elsevierStyleCrossRef" href="#bib0160">32</a> The effect of garlic on the adhesion molecules may partially relate to the inhibition of cytokines.In conclusion, the present data demonstrates that garlic had a protective effect in the imbalance of serum TNF-alpha and IL-4 levels, leukocytes proliferation and liver histological architecture.Conflict of interestThe authors have no conflict of interest to declare." "textoCompletoSecciones" => array:1 [ "secciones" => array:9 [ 0 => array:2 [ "identificador" => "xres86138" "titulo" => array:5 [ 0 => "Abstract" 1 => "Background and aims" 2 => "Methods" 3 => "Results" 4 => "Conclusions" ] ] 1 => array:2 [ "identificador" => "xpalclavsec74297" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:6 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Reagents" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Experimental animals" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Cytokines in serum" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Leukocytes differential count" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Histopathological examination" ] 5 => array:2 [ "identificador" => "sec0040" "titulo" => "Statistical analysis" ] ] ] 4 => array:3 [ "identificador" => "sec0045" "titulo" => "Results" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0050" "titulo" => "Estimation of TNF-α" ] 1 => array:2 [ "identificador" => "sec0055" "titulo" => "Estimation of IL-4" ] 2 => array:2 [ "identificador" => "sec0060" "titulo" => "Leukocyte differential count" ] 3 => array:2 [ "identificador" => "sec0065" "titulo" => "Histopathological findings" ] ] ] 5 => array:2 [ "identificador" => "sec0070" "titulo" => "Discussion" ] 6 => array:2 [ "identificador" => "sec0075" "titulo" => "Conflict of interest" ] 7 => array:2 [ "identificador" => "xack31895" "titulo" => "Acknowledgements" ] 8 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2011-08-17" "fechaAceptado" => "2011-10-11" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec74297" "palabras" => array:6 [ 0 => "Acute hepatic failure" 1 => "D-GalN" 2 => "LPS" 3 => "TNF-alpha" 4 => "IL-4" 5 => "Leukocytes" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "Background and aimsActivation of the pro-inflammatory and anti-inflammatory cytokine cascade, including tumour necrosis factor (TNF)-alpha and interleukin (IL)-4, is considered to play an important role in severe liver injury. Kupffer cells, resident macrophages of the liver, activated with lipopolysaccharide (LPS) release pro-inflammatory cytokine. d-Galactosamine (d-GalN), a hepatocyte-specific inhibitor of RNA synthesis, is known to sensitise animals to the lethal effects of LPS. In the present study we seek to reverse some altered parameters, immunological and histopathological, to normal values of rats pre-treated with garlic. MethodsAcute hepatic failure was induced in male albino rats by the intraperitoneal injection of 500mg d-GalN and 50μg LPS/kg body weight. Expression levels of TNF-α and IL-4 were detected by ELISA. Leukocytes proliferation was carried out by differential count. For histopathology, liver sections were stained with haematoxylin and eosin. Data were analysed by SPSS program version 13.0. ResultsThe data showed significant increase in the numbers of granulocytes, but with significant decreases in lymphocyte and monocytes proliferation and the TNF-alpha and IL-4 levels in d-GalN/LPS-induced group. Garlic pre-treatment of liver-injured rats induced significant amelioration in the numbers of monocytes and lymphocytes, with significant increase in granulocytes numbers, TNF-α level and IL-4 level. ConclusionsResults of this study revealed that garlic could afford a significant protection in the alleviation of d-GalN/LPS-induced hepatocellular injury." ] ] "multimedia" => array:4 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1182 "Ancho" => 2103 "Tamanyo" => 139409 ] ] "descripcion" => array:1 [ "en" => "Changes in TNF-α level in d-GalN/LPS injected groups versus control group and garlic treated groups. Significant differences between all groups, P<0.05, were monitored." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1230 "Ancho" => 2090 "Tamanyo" => 139052 ] ] "descripcion" => array:1 [ "en" => "Changes in IL-4 levels in d-GalN/LPS injected groups versus control group and garlic treated groups. Significant differences between all groups, P<0.05, were monitored." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 1562 "Ancho" => 1750 "Tamanyo" => 420361 ] ] "descripcion" => array:1 [ "en" => "Light photomicrographs of rat liver (haematoxylin and eosin stains, magnification 100×): (A) Control rat liver section (B) d-GalN/LPS-intoxicated rat liver section; (C) liver section of rat pre-treated with garlic." ] ] 3 => array:7 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:3 [ "leyenda" => "Values expressed are mean±SE of five samples." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Groups \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Granulocytes \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Lymphocytes \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Monocytes \t\t\t\t\t\t\n \t\t\t\t</td></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1. Negative control, untreated \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">57.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.40±0.244 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.40±0.244 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2. Positive control, PBS \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">57.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.80±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.20±0.200 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3. Garlic 1 W before \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">57.60±0.509 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.20±0.200 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">4. d-GalN/LPS, 6h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9.60±0.678 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5. d-GalN/LPS/garlic, 6h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.00±0.707 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.00±0.316 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.00±0.447 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">6. d-GalN/LPS, 12h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">63.80±0.663<a class="elsevierStyleCrossRef" href="#tblfn0005">*</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">30.20±0.663<a class="elsevierStyleCrossRef" href="#tblfn0005">*</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">6.00±0.316<a class="elsevierStyleCrossRef" href="#tblfn0005">*</a> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7. d-GalN/LPS/garlic, 12h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">53.20±0.663 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">32.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">14.60±0.400 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8. d-GalN/LPS, 24h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.00±0.707 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">34.60±0.509 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.40±0.600 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9. d-GalN/LPS/garlic, 24h \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">53.80±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">29.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.00±0.447 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10. d-GalN/LPS, 1 W after \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.80±0.583 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">32.80±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">11.40±0.244 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">11. d-GalN/LPS/garlic, 1 W after \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">53.80±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">29.20±0.374 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.00±0.447 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">12. Garlic, 1 W after \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">57.60±0.509 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">35.40±0.244 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.00±0.316 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab165544.png" ] ] ] "notaPie" => array:1 [ 0 => array:3 [ "identificador" => "tblfn0005" "etiqueta" => "*" "nota" => "Statistical significance (P<0.05) is shown on comparing infected or treated samples versus the control ones." ] ] ] "descripcion" => array:1 [ "en" => "Changes of granulocytes, lymphocytes and monocytes percent in d-GalN/LPS injected group versus control groups and garlic treated groups. 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Trautwein" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "Am J Physiol" "fecha" => "1998" "volumen" => "275" "paginaInicial" => "387" "paginaFinal" => "392" ] ] ] ] ] ] ] ] ] ] "agradecimientos" => array:1 [ 0 => array:3 [ "identificador" => "xack31895" "titulo" => "Acknowledgements" "texto" => "I thank Prof. Dr. Mohamed H.I. (Zoology Department, Faculty of Science, El-Minia University, El-Minia, Egypt) for capturing the histological figures. I also thank Dr. Shaban H.A. (Zoology Department, Faculty of Science, El-Minia University, El-Minia, Egypt) for his efforts in data analysis." ] ] ] "idiomaDefecto" => "en" "url" => "/03010546/0000004000000004/v1_201304101110/S0301054611003648/v1_201304101110/en/main.assets" "Apartado" => array:4 [ "identificador" => "5554" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Original articles" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/03010546/0000004000000004/v1_201304101110/S0301054611003648/v1_201304101110/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054611003648?idApp=UINPBA00004N"]

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Original article

Bahaa K.A. Abdel-Salam

Corresponding author

Manar_Muhamad@yahoo.com

Corresponding author.

, Abd-Alla A.A. Sayed

Zoology Department, Faculty of Science, Minia University, El-Minia 61519, Egypt

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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Lipopolysaccharide &#40;LPS&#41;&#44; a component of the cell wall of Gram-negative bacteria&#44; is generally considered to be a major pathogenic element in Gram-negative bacterial infection&#44; and can eventually cause systemic inflammatory response syndrome&#46;<a class="elsevierStyleCrossRef" href="#bib0005"><span class="elsevierStyleSup">1</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">The innate immune system copes with infection by producing pro-inflammatory mediators such as tumour necrosis factor-&#945; &#40;TNF-&#945;&#41;&#59; however&#44; overproduction of these mediators eventually leads to systemic inflammatory response syndrome&#46;<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> LPS stimulates monocytes&#47;macrophages via Toll-like receptor 4 &#40;TLR4&#41;&#44; which belongs to the TLR family&#46;<a class="elsevierStyleCrossRef" href="#bib0015"><span class="elsevierStyleSup">3</span></a> LPS is recognised by TLR4 together with accessory molecules such as myeloid differentiation protein-2 and CD14&#44; and TLR4 then transduces LPS signalling via both myeloid differentiation factor 88 &#40;MyD88&#41;-dependent and MyD88-independent pathways&#44; each of which activates nuclear factor-&#954;B &#40;NF-&#954;B&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleSmallCaps">d</span>-Galactosamine &#40;<span class="elsevierStyleSmallCaps">d</span>-GalN&#41; increases the susceptibility of mice to LPS-induced shock by impairing liver metabolism&#46;<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a> In contrast to high dose LPS-induced shock which induces a systemic disorder including multiple organ failures&#44;<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a> liver is a major target organ after challenge with low doses of LPS in conjunction with <span class="elsevierStyleSmallCaps">d</span>-GalN&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> Similarly to high dose LPS-induced shock&#44; TNF-&#945; plays a central role in low dose LPS-induced shock&#47;liver injury&#46;<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">Endotoxin makes macrophages&#44; neutrophils&#44; and other inflammatory cells release cytokines&#44; such as TNF-&#945;&#44; in great amounts&#44; which may render inflammatory reactions out of control&#44; ultimately resulting in sepsis&#44; septic shock&#44; or multiple organ failure syndromes&#46;<a class="elsevierStyleCrossRefs" href="#bib0010"><span class="elsevierStyleSup">2&#44;9</span></a> The release of TNF-&#945; is the hallmark of the cellular response to the activation of the innate immune system&#46;<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> The essential role of TNF in control of several infections was documented&#46;<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a></p><p id="par0025" class="elsevierStylePara elsevierViewall">A critical degree of liver cell death due to apoptosis or necrosis is fundamental to the development of fulminant hepatic failure&#46;<a class="elsevierStyleCrossRef" href="#bib0060"><span class="elsevierStyleSup">12</span></a> Activation of the pro-inflammatory cytokine cascade&#44; including TNF-alpha is considered to play an important role in the pathophysiology and clinical outcome of this severe liver injury&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a> Kupffer cells&#44; resident macrophages of the liver&#44; have a transmembrane protein TLR-4&#44; which recognises LPS and mediates macrophage activation and pro-inflammatory cytokine release&#46;<a class="elsevierStyleCrossRefs" href="#bib0070"><span class="elsevierStyleSup">14&#44;15</span></a> Upregulated TLR4 expression and function was observed in various situations of experimental liver injury&#46;<a class="elsevierStyleCrossRefs" href="#bib0075"><span class="elsevierStyleSup">15&#8211;18</span></a> Liver macrophages are reported to be increased in number and considered to be closely related to the upregulation of cytokines and chemokines in fulminant hepatic failure&#46;<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">19</span></a></p><p id="par0030" class="elsevierStylePara elsevierViewall">Garlic is a sulphur-rich phytomedicine with a long history of therapeutic use&#46; It has been shown to be useful as an antiplatelet&#44;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">20</span></a> antihypertensive&#44;<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">21</span></a> antifungal&#44;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">22</span></a> anticancer&#44;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">23</span></a> antioxidant&#44;<a class="elsevierStyleCrossRef" href="#bib0120"><span class="elsevierStyleSup">24</span></a> hypolipidaemic<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">25</span></a> and hypoglycaemic&#46;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">26</span></a> Its variety of therapeutic actions probably relates to the complex nature of the chemistry of garlic&#46; It contains many sulphur-rich derivatives of the amino acid cysteinedalliin&#46; The immunomodulatory effects of garlic have been demonstrated on a small variety of cells and tissues&#46; The effects of garlic on human and murine peripheral blood mononuclear cells &#40;PMBCs&#41; and neutrophils have been investigated&#46; Although not investigating the effect of garlic on cytokine production&#46; Therefore&#44; the aim of the present study is to examine the in vivo effect of garlic on normal and <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS treated liver tissues&#46; We directed our efforts at the effect of garlic on the inflammatory cytokine balance of the liver tissue&#46; We wished to examine garlic&#39;s potential to increase anti-inflammatory&#44; and decrease inflammatory mediator production&#44; and thus explore its use as a substance with the potential to enhance or correct liver functions&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Reagents</span><p id="par0035" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleSmallCaps">d</span>-GalN and LPS were obtained from Sigma &#40;St&#46; Louis&#44; MO&#44; USA&#41;&#46; ELISA kits were purchased from R&#38;D Systems &#40;Minneapolis&#44; MN&#44; USA&#41;&#46; All other chemicals were obtained commercially as reagent-grade products&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Experimental animals</span><p id="par0040" class="elsevierStylePara elsevierViewall">Male albino rats&#44; <span class="elsevierStyleItalic">Rattus norvegicus</span> &#40;6&#8211;8 weeks old&#44; weight 100&#8211;120<span class="elsevierStyleHsp" style=""></span>g&#41; were purchased from the Biological Supply Center&#44; Theodore Bilharz Research Institute&#44; TBRI&#44; Cairo&#44; Egypt and housed under specific pathogen-free conditions and maintained on a 12-h light&#8211;dark cycle&#44; with food and water ad libitum&#46; Animals were classified into 12 groups &#40;ten animals each&#41;&#46; After rats were fasted for 18<span class="elsevierStyleHsp" style=""></span>h&#44; the first group &#40;negative control&#41; was untreated&#59; the second group &#40;first positive control&#41; was injected with the same amount of saline intraperitoneally&#59; the third group &#40;second positive control&#41; was allowed free to feed on a crude garlic used as a food supplement &#40;5<span class="elsevierStyleHsp" style=""></span>g&#41; daily for one week before <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection&#46; The groups from four to eleven were injected intraperitoneally with <span class="elsevierStyleSmallCaps">d</span>-GalN at 500<span class="elsevierStyleHsp" style=""></span>mg&#47;kg and LPS at 50<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;kg &#40;dissolved in 500<span class="elsevierStyleHsp" style=""></span>ml of saline&#41; to induce liver injury&#59; the fifth&#44; seventh&#44; ninth and twelfth groups were allowed free to feed on garlic daily for one week after <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection&#46; Blood samples were collected from tail at 6&#44; 12&#44; and 24<span class="elsevierStyleHsp" style=""></span>h after <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection and at one week before and one week after <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Cytokines in serum</span><p id="par0045" class="elsevierStylePara elsevierViewall">Seventy-two hours after the last treating&#44; 10 animals from each group were sacrificed under chloroform anaesthesia&#46; Blood samples were taken and centrifuged at 3000<span class="elsevierStyleHsp" style=""></span>rpm for 30<span class="elsevierStyleHsp" style=""></span>min&#46; Sera were removed and kept at &#8722;20<span class="elsevierStyleHsp" style=""></span>&#176;C for the estimation of TNF-&#945; and IL-4 levels&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Leukocytes differential count</span><p id="par0050" class="elsevierStylePara elsevierViewall">Freshly collected blood samples of about 20<span class="elsevierStyleHsp" style=""></span>&#956;l were spread on clean slides into a thin film using another smooth-edged glass slide&#46; Each blood smear was left to air dry before being fixed with methanol for 2&#8211;3<span class="elsevierStyleHsp" style=""></span>min and then labelled by the number of the animal&#46; Blood smears were stained with 10&#37; Giemsa&#39;s stain &#40;Aldrich&#41; in buffered distilled water containing 0&#46;021<span class="elsevierStyleHsp" style=""></span>M Na<span class="elsevierStyleInf">2</span>HPO<span class="elsevierStyleInf">4</span>&#47;0&#46;015<span class="elsevierStyleHsp" style=""></span>M KH<span class="elsevierStyleInf">2</span>PO<span class="elsevierStyleInf">4</span>&#46; pH 7&#8211;7&#46;2 for 30<span class="elsevierStyleHsp" style=""></span>min far from sun light&#46; After that&#44; the stain was removed by gentle washing with distilled water and the slides were air-dried at room temperature&#46; Using light microscopy at 400&#215; magnification&#44; different types of blood leukocytes were recorded&#46; At least double smears for each blood samples were counted&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Histopathological examination</span><p id="par0055" class="elsevierStylePara elsevierViewall">Liver samples were dissected after DGalN&#47;LPS injection and garlic treating&#44; fixed in 10&#37; formal saline&#44; embedded in paraffin&#44; and cut into 5<span class="elsevierStyleHsp" style=""></span>&#956;m thickness in microtome&#46; These sections were collected in slides and then stained with haematoxylin and eosin&#46; The stained sections were examined under low and high power by using an Olympus microscope&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Statistical analysis</span><p id="par0060" class="elsevierStylePara elsevierViewall">Data were analysed using SPSS program version 13&#46;0&#46; Statistical analysis of the obtained data was performed using one way analysis of variance &#40;ANOVA&#41; test followed by least square differences &#40;LSD&#41; analysis for comparison between means&#46; Results were expressed as mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard error &#40;SE&#41;&#46; Values of <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05 were considered statistically significant&#44; while value of <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05 were considered statistically non-significant&#46;</p></span></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Results</span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Estimation of TNF-&#945;</span><p id="par0065" class="elsevierStylePara elsevierViewall">Serum TNF-&#945; levels were demonstrated in <a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>&#44; where the negative control group &#40;1405&#46;26<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>5&#46;252<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; the first PBS positive control &#40;1431&#46;59<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>10&#46;692<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and the second positive control pre-treated with garlic one week before <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection &#40;1361&#46;48<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>8&#46;302<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; is higher than the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS treated group after 6<span class="elsevierStyleHsp" style=""></span>h &#40;172&#46;16<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;049<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; 12<span class="elsevierStyleHsp" style=""></span>h &#40;880&#46;87<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;717<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and 24<span class="elsevierStyleHsp" style=""></span>h &#40;969&#46;73<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;114<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; which is lower than the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS group treated with garlic &#40;1129&#46;73<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;650<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 1726&#46;02<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>4&#46;376<span class="elsevierStyleHsp" style=""></span>pg&#47;ml and 1561&#46;90<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>6&#46;574<span class="elsevierStyleHsp" style=""></span>pg&#47;ml at 6&#44; 12 and 24<span class="elsevierStyleHsp" style=""></span>h&#44; respectively&#41;&#46; In contrast&#44; the garlic treated group after one week of <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection &#40;1466&#46;51<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;451<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; is higher than the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups at 6&#44; 12 and 24<span class="elsevierStyleHsp" style=""></span>h &#40;172&#46;16<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;049<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 880&#46;87<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;717<span class="elsevierStyleHsp" style=""></span>pg&#47;ml and 969&#46;73<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;114<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; respectively&#41;&#46; These results showed a significant difference in TNF-&#945; level between all groups &#40;<span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Estimation of IL-4</span><p id="par0070" class="elsevierStylePara elsevierViewall">Results of ELISA in <a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a> showed that IL-4 recorded 667&#46;72<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>7&#46;270<span class="elsevierStyleHsp" style=""></span>pg&#47;ml in negative control group &#40;untreated&#41;&#44; 640&#46;09<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;176<span class="elsevierStyleHsp" style=""></span>pg&#47;ml in the first PBS positive control and 687&#46;00<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>4&#46;078<span class="elsevierStyleHsp" style=""></span>pg&#47;ml in the second positive control&#46; Liver injury group injected with <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS counted low levels of IL-4 at 6<span class="elsevierStyleHsp" style=""></span>h &#40;283&#46;50<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;417<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; 12<span class="elsevierStyleHsp" style=""></span>h &#40;368&#46;10<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;607<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and 24<span class="elsevierStyleHsp" style=""></span>h &#40;454&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;994<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; than the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups treated with garlic &#40;555&#46;75<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;864<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; 813&#46;59<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>2&#46;081<span class="elsevierStyleHsp" style=""></span>pg&#47;ml and 769&#46;63<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;421<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; at the same time&#44; respectively&#41;&#46; In addition&#44; the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups at 6&#44; 12 and 24<span class="elsevierStyleHsp" style=""></span>h are lower than the garlic treated group after one week of <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection &#40;751&#46;01<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;789<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#46; Statistical analysis showed that there is a significant difference in IL-4 between the liver injury groups and either the control group or the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups treated with garlic&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Leukocyte differential count</span><p id="par0075" class="elsevierStylePara elsevierViewall"><a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a> shows that granulocytes percentage in the garlic treated groups before &#40;57&#46;60<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;509<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; and after &#40;57&#46;60<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;509<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injection&#44; the negative control group &#40;57&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;374<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#44; the first positive control group treated saline &#40;57&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;374<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41; are nearly similar in population and higher than the other groups ranged from 53&#46;80<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;374<span class="elsevierStyleHsp" style=""></span>pg&#47;ml to 55&#46;80<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;583<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; except for the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected group after 12<span class="elsevierStyleHsp" style=""></span>h which showed the highest population &#40;63&#46;80<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;663<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#41;&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0080" class="elsevierStylePara elsevierViewall">In contrast to granulocytes&#44; the lymphocytes and monocytes percentages showed the lowest population in the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected group after 12<span class="elsevierStyleHsp" style=""></span>h &#40;30&#46;20<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;663<span class="elsevierStyleHsp" style=""></span>pg&#47;ml and 6&#46;00<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;316<span class="elsevierStyleHsp" style=""></span>pg&#47;ml&#44; respectively&#41;&#44; <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Statistical analysis showed that there is a significant difference in leukocytes counts between the liver injury groups and either the control group or the groups treated with garlic&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#46;</p></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Histopathological findings</span><p id="par0090" class="elsevierStylePara elsevierViewall">Histopathological examination of liver sections from control animals revealed normal architecture&#44; <a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>A&#46; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS-intoxicated rat liver section showed loss of architecture and prominent inflammatory collections in the central rather than around central vein&#44; <a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>B&#46; In contrast&#44; a slight inflammatory reaction was found in liver section of rat pre-treated with garlic prior to <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS challenge&#44; <a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>C&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Discussion</span><p id="par0095" class="elsevierStylePara elsevierViewall">Many approaches to suppress the effects of inflammatory mediators have been unsatisfactory for the effective treatment of Gram-negative bacterial&#46;<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> This promoted us to use a medicinal plant&#44; garlic&#44; to treat bacterial infection causing liver injury&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">This paper describes participation of <span class="elsevierStyleSmallCaps">d</span>-GalN and LPS in liver injury and points to garlic as a critical medicinal plant of resistance to this injury&#46; Histological disorder is accompanied with the level of some cytokines&#44; such as the pro-inflammatory TNF-&#945; and the anti-inflammatory IL-4&#44; and leukocytes proliferation&#46;</p><p id="par0105" class="elsevierStylePara elsevierViewall">The liver is an important organ within the body with a central role in metabolic homeostasis&#44; as it is responsible for the metabolism&#44; synthesis&#44; storage&#44; and redistribution of nutrients&#44; carbohydrates&#44; fats&#44; and vitamins&#46; Hepatitis is the best-known liver disease&#46;</p><p id="par0110" class="elsevierStylePara elsevierViewall">Acute liver injury was induced by <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> The advantage of this induction is that <span class="elsevierStyleSmallCaps">d</span>-GalN can potentiate the toxic effects of LPS and produce fulminant hepatitis within a few hours&#46;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">27</span></a> In the present study&#44; we demonstrated that pre-treatment with garlic attenuated liver injury caused <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#44; where the administration of <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS significantly down-regulated the expression of both serum TNF-alpha and IL-4 that are up-regulated with garlic&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">NF-kappa B plays a key role in the regulation of TNF-alpha transcription stimulated by LPS&#46;<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">28</span></a> In the latent state&#44; NF-kappa B is found in cytoplasm&#46; After exposure to LPS&#44; NF-kappa B is translocated to the nucleus and binds to NF-kappa B promoter sites on DNA&#44; activating gene transcription of cytokines such as TNF-alpha&#46;<a class="elsevierStyleCrossRef" href="#bib0145"><span class="elsevierStyleSup">29</span></a></p><p id="par0120" class="elsevierStylePara elsevierViewall">Taken together&#44; our results revealed that the up-regulation of TNF-alpha of garlic against <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS-induced liver injury in rat could be related to inhibiting the activation of NF-kappa B&#46; In addition&#44; the down-regulation of the TNF-alpha might also be due to the death of the Kupffer cells&#44; resident macrophages of the liver&#44; where they have a transmembrane protein Toll-like receptor 4 &#40;TLR4&#41;&#44; which recognises endotoxin &#40;LPS&#41; and mediates macrophage activation and pro-inflammatory cytokine release&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a></p><p id="par0125" class="elsevierStylePara elsevierViewall">The histological injuries such as inflammation induced by <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS were markedly improved by garlic&#44; which showed a protective effect as a control&#46; Previous studies showed that LPS exerted the toxic effects mainly through the generation of endogenous inflammatory cytokines secreted from macrophages&#46; Among these cytokines&#44; TNF-&#945; is a key factor that contributes to the triggering of an inflammatory cascade involving the induction of cytokines including interferon-&#947;&#44; nitric oxide&#44; IL-1&#946; and cell adhesion molecules&#44; etc&#46;<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">30</span></a></p><p id="par0130" class="elsevierStylePara elsevierViewall">A proper balance between pro- and anti-inflammatory mediators is necessary for modulating an adequate immune response toward LPS&#46; IL-4 is known to be an anti-inflammatory cytokine which can suppress serum TNF-&#945; levels and reduce the mortality of mice exposed to <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#46;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">31</span></a> In the present study&#44; there was a massive induction of the inflammatory cytokines TNF-&#945; in hepatic failure caused by <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#44; which were not counterbalanced by the anti-inflammatory cytokine IL-4&#46; This imbalance may promote inflammatory responses to liver damage&#46; After administration of garlic&#44; serum IL-4 maintained at a higher level&#44; which indicated that the imbalance was reversed by garlic to a certain extent&#46;</p><p id="par0135" class="elsevierStylePara elsevierViewall">Interaction between T-cells and hepatocytes that was regulated by adhesion molecules is dysregulated in harmful inflammatory processes&#46; During the <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS intoxication&#44; the expressions of ICAM-1 and LFA-1 in hepatocytes and non-parenchymal cells were enhanced&#46; The elevation of these molecules was essential to trigger the extravasations of neutrophils into the liver parenchyma&#44; which produced cytotoxic damage to hepatocytes&#46;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">32</span></a> This is in accordance with the high percentage of neutrophils monitored in our results&#46; Previous studies suggested that TNF-&#945; could induce the transcription of adhesion molecules&#46;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">32</span></a> The effect of garlic on the adhesion molecules may partially relate to the inhibition of cytokines&#46;</p><p id="par0140" class="elsevierStylePara elsevierViewall">In conclusion&#44; the present data demonstrates that garlic had a protective effect in the imbalance of serum TNF-alpha and IL-4 levels&#44; leukocytes proliferation and liver histological architecture&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle">Conflict of interest</span><p id="par0145" class="elsevierStylePara elsevierViewall">The authors have no conflict of interest to declare&#46;</p></span></span>"
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              "titulo" => "Cytokines in serum"
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              "titulo" => "Leukocytes differential count"
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              "titulo" => "Estimation of TNF-&#945;"
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              "titulo" => "Estimation of IL-4"
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            3 => array:2 [
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        5 => array:2 [
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      "en" => array:2 [
        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle">Background and aims</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Activation of the pro-inflammatory and anti-inflammatory cytokine cascade&#44; including tumour necrosis factor &#40;TNF&#41;-alpha and interleukin &#40;IL&#41;-4&#44; is considered to play an important role in severe liver injury&#46; Kupffer cells&#44; resident macrophages of the liver&#44; activated with lipopolysaccharide &#40;LPS&#41; release pro-inflammatory cytokine&#46; <span class="elsevierStyleSmallCaps">d</span>-Galactosamine &#40;<span class="elsevierStyleSmallCaps">d</span>-GalN&#41;&#44; a hepatocyte-specific inhibitor of RNA synthesis&#44; is known to sensitise animals to the lethal effects of LPS&#46; In the present study we seek to reverse some altered parameters&#44; immunological and histopathological&#44; to normal values of rats pre-treated with garlic&#46;</p> <span class="elsevierStyleSectionTitle">Methods</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Acute hepatic failure was induced in male albino rats by the intraperitoneal injection of 500<span class="elsevierStyleHsp" style=""></span>mg <span class="elsevierStyleSmallCaps">d</span>-GalN and 50<span class="elsevierStyleHsp" style=""></span>&#956;g LPS&#47;kg body weight&#46; Expression levels of TNF-&#945; and IL-4 were detected by ELISA&#46; Leukocytes proliferation was carried out by differential count&#46; For histopathology&#44; liver sections were stained with haematoxylin and eosin&#46; Data were analysed by SPSS program version 13&#46;0&#46;</p> <span class="elsevierStyleSectionTitle">Results</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">The data showed significant increase in the numbers of granulocytes&#44; but with significant decreases in lymphocyte and monocytes proliferation and the TNF-alpha and IL-4 levels in <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS-induced group&#46; Garlic pre-treatment of liver-injured rats induced significant amelioration in the numbers of monocytes and lymphocytes&#44; with significant increase in granulocytes numbers&#44; TNF-&#945; level and IL-4 level&#46;</p> <span class="elsevierStyleSectionTitle">Conclusions</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Results of this study revealed that garlic could afford a significant protection in the alleviation of <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS-induced hepatocellular injury&#46;</p>"
      ]
    ]
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          "en" => "<p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Changes in TNF-&#945; level in <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups versus control group and garlic treated groups&#46; Significant differences between all groups&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; were monitored&#46;</p>"
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      1 => array:7 [
        "identificador" => "fig0010"
        "etiqueta" => "Figure 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
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            "Tamanyo" => 139052
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          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Changes in IL-4 levels in <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS injected groups versus control group and garlic treated groups&#46; Significant differences between all groups&#44; <span class="elsevierStyleItalic">P</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; were monitored&#46;</p>"
        ]
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      2 => array:7 [
        "identificador" => "fig0015"
        "etiqueta" => "Figure 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
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            "Tamanyo" => 420361
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          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Light photomicrographs of rat liver &#40;haematoxylin and eosin stains&#44; magnification 100&#215;&#41;&#58; &#40;A&#41; Control rat liver section &#40;B&#41; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS-intoxicated rat liver section&#59; &#40;C&#41; liver section of rat pre-treated with garlic&#46;</p>"
        ]
      ]
      3 => array:7 [
        "identificador" => "tbl0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "tabla" => array:3 [
          "leyenda" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Values expressed are mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>SE of five samples&#46;</p>"
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                  \t\t\t\t">3&#46; Garlic 1 W before&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">4&#46; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#44; 6<span class="elsevierStyleHsp" style=""></span>h&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">5&#46; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#47;garlic&#44; 6<span class="elsevierStyleHsp" style=""></span>h&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">6&#46; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#44; 12<span class="elsevierStyleHsp" style=""></span>h&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">9&#46; <span class="elsevierStyleSmallCaps">d</span>-GalN&#47;LPS&#47;garlic&#44; 24<span class="elsevierStyleHsp" style=""></span>h&nbsp;\t\t\t\t\t\t\n
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  ]
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  "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054611003648?idApp=UINPBA00004N"
]

Article information

ISSN: 03010546
Original language: English

DOI:10.1016/j.aller.2011.10.001

The statistics are updated each day

Year/Month	Html	Pdf	Total

2024 November	2	1	3
2024 October	16	8	24
2024 September	19	7	26
2024 August	14	7	21
2024 July	10	10	20
2024 June	12	14	26
2024 May	8	6	14
2024 April	17	10	27
2024 March	20	10	30
2024 February	9	4	13
2024 January	15	7	22
2023 December	14	9	23
2023 November	11	9	20
2023 October	10	14	24
2023 September	20	8	28
2023 August	16	10	26
2023 July	13	9	22
2023 June	24	3	27
2023 May	29	4	33
2023 April	10	2	12
2023 March	11	3	14
2023 February	22	3	25
2023 January	20	13	33
2022 December	21	5	26
2022 November	13	6	19
2022 October	13	7	20
2022 September	25	15	40
2022 August	15	8	23
2022 July	14	9	23
2022 June	12	4	16
2022 May	16	10	26
2022 April	14	8	22
2022 March	13	9	22
2022 February	34	9	43
2022 January	25	6	31
2021 December	18	17	35
2021 November	11	8	19
2021 October	31	12	43
2021 September	19	17	36
2021 August	12	7	19
2021 July	14	9	23
2021 June	16	11	27
2021 May	25	13	38
2021 April	30	27	57
2021 March	18	15	33
2021 February	16	14	30
2021 January	7	11	18
2020 December	0	7	7
2020 November	0	2	2
2020 October	0	2	2
2020 September	0	6	6
2020 August	0	2	2
2020 July	0	3	3
2020 June	0	2	2
2020 May	0	14	14
2020 April	0	4	4
2020 March	0	7	7
2020 February	0	13	13
2020 January	0	14	14
2019 December	0	9	9
2019 November	0	1	1
2019 October	0	2	2
2019 September	0	4	4
2019 August	0	7	7
2019 July	0	11	11
2019 June	0	9	9
2019 May	0	68	68
2019 April	0	19	19
2019 March	0	1	1
2019 February	0	2	2
2019 January	0	3	3
2018 August	0	1	1
2018 February	3	2	5
2018 January	7	2	9
2017 December	3	2	5
2017 November	12	6	18
2017 October	5	2	7
2017 September	11	5	16
2017 August	11	4	15
2017 July	6	2	8
2017 June	10	11	21
2017 May	18	8	26
2017 April	10	14	24
2017 March	8	56	64
2017 February	9	5	14
2017 January	13	3	16
2016 December	14	8	22
2016 November	16	4	20
2016 October	25	10	35
2016 September	21	7	28
2016 August	12	7	19
2016 July	5	3	8
2016 June	13	5	18
2016 May	11	8	19
2016 April	13	15	28
2016 March	13	18	31
2016 February	13	19	32
2016 January	10	16	26
2015 December	19	17	36
2015 November	11	19	30
2015 October	15	12	27
2015 September	21	6	27
2015 August	7	4	11
2015 July	4	2	6
2015 June	1	3	4
2015 May	4	9	13
2015 April	5	11	16
2015 March	11	10	21
2015 February	3	9	12
2015 January	16	4	20
2014 December	21	4	25
2014 November	10	7	17
2014 October	26	2	28
2014 September	22	5	27
2014 August	7	2	9
2014 July	8	1	9
2014 June	11	2	13
2014 May	6	4	10
2014 April	6	4	10
2014 March	43	17	60
2014 February	46	11	57
2014 January	58	10	68
2013 December	42	10	52
2013 November	35	5	40
2013 October	43	11	54
2013 September	49	10	59
2013 August	41	8	49
2013 July	42	8	50
2013 June	23	4	27
2013 May	32	7	39
2013 April	25	11	36
2013 March	27	6	33
2013 February	12	8	20
2013 January	12	1	13
2012 December	7	3	10
2012 November	2	3	5
2012 October	2	5	7
2012 September	3	2	5
2012 June	426	0	426

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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