Objective: To probe into the mechanism and interventional effects of silybin-phospholipid complex on amiodarone-induced steatosis in mice. Methods: Eight-week-old male C57BL/6 mice were divided into three groups (5 mice in each group): a control group (WT) with normal diet, a model group with amiodarone 150mg/kg/d by oral gavage (AM), and an intervention group on amiodarone 150mg/kg/d combined with silybin-phospholipid complex(AM+SILIPHOS. All mice were fed their assigned diet for one week. Then, one week later, serum alanine aminotransferase, aspartate aminotransferase, triglyceride, total cholesterol and high-density lipoprotein were detected of each group. A liver pathological change was observed by oil red O and H&E staining. Ultrastructural pathological changes of hepatocytes were observed to evaluate the intervention effect by transmission electron microscopy. RT-q PCR was used to detect the expression of peroxisome proliferator-activated receptor alpha and its regulated lipid metabolism genes CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 in liver tissues. Intra-group comparison was done by paired t-test. One-way ANOVA was used for comparison between groups and semi-quantitative data were tested using Mann-Whitney U test. Results: Oil Red O and H&E staining results of liver tissue in the intervention group showed that intrahepatic steatosis was significantly reduced when compared to model group. Transmission electron microscopy showed that the model group had pyknotic nuclei, mitochondrial swelling, structural damage, and lysosomal degradation whereas the intervention group had hepatic nucleus without pyknosis, reduced mitochondrial swelling and slight structural damage than that of model group. RT-q PCR results showed that the expression of peroxisome proliferator-activated receptor alpha, CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 were increased in the model group but the expression of CPTI, Cyp4a14, Acot1 and peroxisome proliferator-activated receptor alpha were decreased in the intervention group (P < 0.05). Conclusion: Silybin-phospholipid complex can alleviate amiodarone-induced steatosis, and its mechanism may play a role in protecting mitochondrial function and regulating fatty acid metabolism. Thus, silybin-phospholipid complex has potential intervention effect on amiodarone-induced fatty liver.
目的： 探讨水飞蓟宾-磷脂复合物对胺碘酮诱导的药物性脂肪变的干预作用及其机制。 方法： 将8周龄雄性C57BL/6J小鼠分为3组，每组5只：正常饮食为对照组（WT组）；150 mg·kg(-1)·d(-1)胺碘酮灌胃7 d为模型组（AM组）；干预治疗组（AM + SILIPHOS组）系胺碘酮造模同时使用水飞蓟宾-磷脂复合物50 mg·kg(-1)·d(-1)灌胃，连续7 d。1周后通过检测各组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，用HE染色和油红染色观察肝脏病理学改变，用透射电镜观察肝细胞的超微病理学变化，评估干预效果；用RT-qPCR检测肝组织内脂肪代谢的主要调节因子过氧化物酶体增殖剂激活受体α及其调节的脂代谢基因CPTI、CPTII、Acot1、Acot2、ACOX、Cyp4a10和Cyp4a14的表达，探索其可能的机制。数据组内比较采用配对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： 干预组肝脏组织油红及HE染色结果显示肝内脂肪变较模型组小鼠明显减少。透射电镜结果显示模型组肝细胞核固缩，线粒体肿胀，结构破损，并被溶酶体自噬；干预组肝细胞核略受损，但无固缩，线粒体肿胀较模型组减轻。RT-qPCR检测结果显示模型组过氧化物酶体增殖物激活受体α、CPTI、CPTII、Acot1、Acot2、ACOX、Cyp4a10、Cyp4a14的mRNA的表达上升，经干预治疗后CPTI、Cyp4a14、Acot1、过氧化物酶体增殖剂激活受体α表达降低（P值均< 0.05）。 结论： 水飞蓟宾-磷脂复合物可减轻胺碘酮引起的药物性脂肪变，其机制可能通过保护线粒体功能和调节脂肪酸代谢起作用，提示水飞蓟宾-磷脂复合物对胺碘酮诱导的药物性脂肪肝具有潜在干预作用。.
Keywords: Amiodarone; Drug-induced liver injury; Fatty liver; Models, animal; Silybin-phospholipid complex.