胺碘酮通过PPARα信号通路诱导肝脏脂肪变的机制研究

Mechanism study of liver steatosis induced by amiodarone via PPARαsignaling pathway

目的探讨胺碘酮诱导的肝脏脂肪变的机制。方法8周雄性C57BL/6J小鼠分为3组,每组5只:正常饮食为空白对照组(WT组,n=5);胺碘酮(150 mg·kg-1·d^(-1))灌胃10 d为模型组(胺碘酮组,n=5);干预治疗组[胺碘酮+PPARα激动剂WY14643(2.5 mg/kg),n=5]即胺碘酮造模同时使用WY14643(2.5 mg/kg)灌胃,连续10 d。10 d后检测小鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、甘油三酯、总胆固醇和高密度脂蛋白,HE染色和油红染色观察肝脏病理学改变;RT-PCR检测肝组织内过氧化物酶体增殖物激活受体α(PPARα)及其调节的脂代谢基因的表达。数据组内比较采用t检验,组间比较用单因素方差分析,半定量资料采用非参数检验的Mann-Whitney U检验。结果WY14643干预治疗组肝组织油红及HE染色结果显示,肝内脂肪变较模型组小鼠明显减少。RT-PCR检测结果显示,胺碘酮组PPARα及下游相关基因CPTI、ACOX、Cyp4a14的mRNA表达均下降,治疗后PPARα及部分相关基因CPTI、ACO、Cyp4a14表达较胺碘酮组上升(P=0.0003,0.0157,0.0057,0.0203)。结论胺碘酮可能通过调控PPARα及其下游相关基因诱发肝脏脂肪变。

Objective To investigate the mechanism of hepatic steatosis induced by amiodarone.Methods Male C57BL/6J mice aged 8 weeks were divided into 3 groups of 5.control group with normal diet(WT group,n=5);Amiodarone at 150mg.kg^(-1).d^(-1)was administered intragastatically for 10 days as the model group(AM group,n=5).In the intervention group(AM+PPARαagonist WY14643(2.5mg/kg),n=5)amiodarone was used simultaneously by gavage with WY14643(2.5mg/kg)for 10 consecutive days;After 10 day,serum alanine aminotransferase,aspartate aminotransferase,triglyceride,total cholesterol and high density lipoprotein in each group were detected.The pathological changes of liver were observed by HE staining and oil red staining.The expression of PPARαand its regulated lipid metabolism genes were detected by RT-PCR.t test was used for intra-group comparison,one-way analysis of variance was used for inter-group comparison,and Mann-Whitney U test for non-parametric test was used for semi-quantitative data.Results Pathological observation showed that compared with the model group,the results of liver oil red and HE staining in the intervention group showed that the liver fat change was significantly reduced;RT-PCR results showed that mRNA expression of PPARαand downstream related genes CPTI,Cyp4a14,Acot1decreased in amiodarone group.After treatment,the expressions of PPARαand some related genes were increased compared with those of model group((P=0.0003,0.0157,0.0057,0.0203).Conclusion Amiodarone may induce liver steatosis by regulating PPARαand its downstream related genes.