Background and aim: Activation of stellar liver cells is the cellular basis for the establishment of liver fibrosis. In search of drugs that reverse and/or inhibit the fibrogenic process, plants are important sources of bioactive compounds. Turnera diffusa methanol extract (METD) shows “hepatoprotective” activity. Aim: To explore the effect of METD on the expression of fibrotic markers, modulators of extracellular matrix proteins (ECM), underlying mitochondrial mechanisms and epithelial-mesenchymal transition (EMT) in an in vitro model of human liver stellar cells (LX-2).
Material and methods: The IC50 of the METD in the LX-2 cells was evaluated by the MTT assay. LX-2 cells were exposed to METD (100-200 ng/mL) with TGF-β (10 ng/mL) at 24, 48 and 72h. RNA and proteins were extracted, RT-PCR, qPCR and WB were performed, the relative expression of tumor growth factor beta (TGF-β), collagen 1α 1 (COL1α-1), smooth muscle alpha actin (α-SMA), inhibitor of metalloproteinase 1 (TIMP1), metalloproteinase 2 (MMP2), SNAIL1 an EMT marker and mitofusin 2 (MNF2) of mitochondrial function. Endogenous β-actin gene and GAPDH. ANOVA analysis (p<0.05).
Results: The METD has a concentration of 150 ng/mL mantain over 80% viability in LX-2 cells. The presence of METD in cells treated with TGF-β modifies fibrogenic markers, decreasing COL1α-1 and increasing α-SMA RNA expression at all times, but increase the translational expression of α-SMA at 48 and 72h. We find TIMP1 and MMP2 RNA overexpression, and decreased TIMP1 translational expression was found at all times. It was found Snail1 and MFN2 RNA overexpression, controversially found decreased the translation of MNF2 at all times.
Conclusions: The METD modulates the expression of pro-fibrogenic markers, ECM modulators and some pathways related to EMT and mitochondrial morphology and function, attenuating the expression of profibrogen markers in human LX-2 stellar cells. This work was partially subsidized by PAICYT SA669-18. Registration number of the ethics committee HI11-003.
Conflicts of interest: The authors have no conflicts of interest to declare.
