MnTMPyP mitigates Time-Dependent Molecular Alterations in Ethanol-Induced Hepatic Stress in HepG2 cells

Abstract

Alcohol associated liver disease (ALD) is a multifactorial disease causes due to persistent intake of alcohol. That entails a complicated interaction among oxidative stress, lipid metabolism, inflammatory pathways, hypoxia regulatory signaling pathways and apoptosis, eventually induces fibrogenesis. The study aims to assess the molecular alteration occurred in ethanol-induced stress associated with biomarkers in ALD pathogenesis and fibrosis in experimental in-vitro model. HepG2 cells in 20 Mm of ethanol concentration were for 1-5 days and the morphology and oxidative stress parameters, lipids levels (TBARS and protein carbonyls), hypoxic markers expression, apoptosis level were estimated 1,3 and 5 days of the experiment. RT-PCR was employed to determine the m-RNA expressions of   CYP1A7, PPAR-α, iNOS, LCAT, SREBP-1c, ABCA1, ABCG1, HIF-1α, HIF-2α, miR-21, miR-26A respectively. The changes in morphology were observed in time dependent manner by ethanol consumption. The results demonstrated ethanol causes progressive membrane damage, and increase in TBARs levels and protein carbonyl, indicating augmented lipid peroxidation. Whereas, the expression of SREBP-1c, iNOS, HIF-1α, HIF-2α, p53, miR-21 and CYP1A7 markedly increased while expression levels of PPAR-γ, ABCA1, ABCG1and miR-26Alevels were significantly suppressed. That suggested the lipogenesis intensification, impaired fatty acid oxidation and cellular cholesterol exportation. Also, inflammatory signaling and hypoxic pathways was triggered, along with the apoptotic and fibrotic process initiation, whereby increased miR-21, while decreased miR-26Aexpression defines the shift toward pro-fibrotic regulatory axis. Hence, these molecular alterations may constitute valid biological marker in predicting the development of ALD and might also be possible targets for therapeutic management of the disease.