Micro RNA-203 Inhibits Alcohol-Induced Hepatic Steatosis By Targeting Lipin1

Aim Alcohol liver disease(ALD)is a global liver disease which characterized by liver inflammation,fatty liver,alcoholic hepatitis,or liver cirrhosis.Alcohol abuse is one of the main reasons for liver disease.Alcoholic fatty liver(AFL)disease is the early stage of ALD and associated with the excessive lipids accumulation in hepatocytes as well as oxidative stress.So reverse the alcoholic fatty liver effective is one of the important measures to prevent the deterioration of ALD.However,satisfactory treatment drugs and methods for AFL are still lack so far.So it is significant to clarify the molecular mechanism of alcoholic fatty liver. MicroRNA-203(miR-203)is known to suppress the proliferation and metastasis of hepatocellular carcinoma,but the role in the progression of alcoholic liver disease is not clear and is warranted for further investigation.Lipin1 is an important member of the Lipin family and it has the regulatory functions shared by the Lipin family.On the one hand,it can promote the conversion of phosphatidic acid into triglyceride,and on the other hand it can form complexes with PPAR-? and regulate the oxidation of fatty acids.Method Current study explore the function of miR-203 and regulate mechanism of lipid metabolism,thus providing a new target for cure of alcoholic fatty liver.Firstly,we construct the alcoholic fatty liver mice model and QPCR detect the expression of miR-203.Then over-expressed miR-203 by lentivirus vector in vivo,HE and Oil red staining were used to observe the pathological changes in the liver of mice.Western blot analysis for protein expression of lipid metabolism markers PPAR-?,SREBP-1,inflammation markers IL-6,TNF-?.To over-expression of miR-203 in vitro,we transfected mi R-203 mimics by Lip2000.The cellular Oil Red staining and cellular TG and TCH levels were used to detect intracellular lipid accumulation.Western blot analysis for protein expression of lipid metabolism markers PPAR-?,SREBP-1,inflammation markers IL-6,TNF-?.Furthermore,bioinformatics analyses and dual luciferase assays proved that miR-203 could bind to Lipin1.Knock down Lipin1 by transfecting Lipin1-siRNA in vitro,western blot analysis for protein expression of lipid metabolism markers and fatty acid synthase.To investigate the effect of miR-203 on subcellular localization of Lipin1,nuclear protein and cytoplasmic protein were extracted after over-expression of miR-203 in vitro.Western blot was used to detect the expression level of Lipin1 and immunohistochemistry was used to detect the subcellular localization of Lipin1.Results This experiment found that expression of miR-203 in alcoholic fatty liver was decreased,and over-expression of miR-203 could inhibit the lipid accumulation of alcoholic fatty liver abnormalities and liver inflammation.Furthermore we found that miR-203 regulated liver lipid metabolism may be related to Lipin1.Knocked down Lipin1 could inhibit lipid abnormal metabolism and expression of fatty acid synthase in liver.This process is related to the subcellular localization of Lipin1.Over-expression of miR-203 can promote Lipin1 nuclei localization and inhibit its cytoplasmic localization.Conclusion miR-203 can inhibit the abnormal lipid metabolism of hepatocytes in alcoholic fatty liver,and this process is related to the targeted regulation of Lipin1.