Effects Of Ethanol On Expression Of PPAR-A、AMPK-A1and Apoptosis In Rat Hepatoma H4-ⅡE Cells

Objective:To observe the effects of ethanol on expression of AMPK-α1and PPAR-a on rat hepatoma cell lines H4-ⅡE.Methods:The apoptosis induced by ethanol on rat hepatoma cell lines H4-ⅡE was detected by inverted microscope,optical microscope,flow cytometry(FCM),TUNEL assay and Annexin V/PI double labeling method.The expression of AMPK-α1and PPAR-α were detected by immunohistochemitry(ICH).Results:1.morphological observation:(1)Inverted microscope observation:the cell counts of0.5M ethanol group was less than that of control group.the number of1.0M and1.5M ethanol group were reduced than the0.5M group.(2)HE staining had shown that there were classical apoptosis changes such as apoptosis bodies in ethanol groups.2.FCM:the counts of living cells were reduced in the ethanol groups(P <0.05).the counts of early apoptotic and late apoptotic cells in ethanol group were increased than the control group(P<0.05).3.Annexin V/PI double labeling:Part of green fluorescent cells and small amount of red fluorescent particles were visible under the fluorescence microscope,and more green and red fluorescent cells were found in ethanol groups.4.TUNEL assay:the counts of blue-stained nucleus in ethanol group were increased than the control group(P<0.05).5.IHC:(1)the expression of PPAR-α was reduced in the ethanol groups but the0.5M12h ethanol group(P<0.05).(2)the expression of AMPK-α1was reduced in the ethanol groups but the0.5M12h ethanol group(P<0.05).Conclusion:1.ethanol reduce the expression of PPAR-α on hepatocytes;2.ethanol reduce the expression of AMPK-α1on hepatocytes;3.ethanol induce apoptosis on hepatocytes is related with PPAR-α downregulation.