Background and aim: Intestinal permeability increases in alcoholics allowing the passage of lipopolysaccharide (LPS) to liver, promoting TLR-4 activation in Kupffer cells and the production of pro-inflammatory cytokines. At the present, the modulation of LPS in alcoholics at systemic level, is not fully understood. Aim. To evaluate TLR-4 in M1 monocytes and the production of IL-6 and CXCL-8 in alcoholic liver disease.
Material and methods: Cross-sectional study, that include Alcoholic patients (according WHO) from the Liver clinic of the General Hospital of Mexico were included. They were classified by absence (OH) or presence (CiOH) of liver damage and patients with active alcoholic hepatitis (HOH). Control group (CT): AUDIT <8 and intake of <10gOH / day. Blood samples were taken on one occasion (10ml) to obtain mononuclear cells by density gradient. To which cell marking was performed by flow cytometry (M1 and TLR-4) and in serum was performed the determination of cytokines (IL-6 and CXCL-8) by arrangement in multiple suspension. Mann Whitney U statistical analysis. All patients signed informed consent. Protocol approved by the General Hospital of Mexico (HG/DI/16/107/03/082) and UNAM (FMD/DI/15/2019).
Results: 24 CT, 12 OH, 10 CiOH and 10 HOH were included. The percentage of Monocytes was: CT=8%, OH=19%, CiOH=22% and HOH=35% (OHvsCT p=0.003, CiOHvsCT p=0.05, HOHvsCT p<0.0019). The significant differences in M1 monocytes were found in CiOHvsCT p=0.05, HOHvsCT p=0.019, CiOHvsOHp=0.009 and HOHvsOH p=0.01. Interestingly, TLR4 expression showed differences in OHvsCT p=0.002, CiOHvsCT p=0.007 and HOHvsCT p<0.001. On the other hand, the concentration of IL-6 and IL-8 (pg/mL) presented significant differences according with liver damage (CiOHvsCT p<0.05, HOHvsCT p<0.001, OHvs.HOH p=0.001 and CiOHvsHOH p=0.01).
Conclusions: Alcohol has an effect at a systemic level promoting the increase of monocytes/M1 and the expression of TLR-4, supporting the fact that the receptor is activated in the periphery, being higher in patients with active Alcoholic Hepatitis, favoring a state of continuous inflammation and promoting susceptibility to infections and mortality.
This work has been partially funded by CONACYT: SALUD-2016-272579.
Conflicts of interest: The authors have no conflicts of interest to declare.
