苦参碱抑制游离脂肪酸诱导小鼠肝实质细胞脂肪变性

摘要

目的：探讨苦参碱对游离脂肪酸（free fatty acids，FFA）诱导的小鼠肝实质细胞脂肪变性的影响。方法Ⅳ型胶原酶灌注分离 BALB／c 小鼠原代肝实质细胞并进行体外培养。实验分对照组（CG），高脂组（FG）及苦参碱组（MG）。通过体外测定细胞内三酰甘油的含量。用油红染色的方法观察肝实质细胞的脂肪样变性。Western 印迹检测细胞内MAPK 相关信号通路磷酸化的变化。实时定量 PCR 检测细胞中与脂肪化密切相关的 miR-122的表达和相关靶基因的表达改变。结果与高脂组比较，苦参碱组肝实质细胞内三酰甘油含量显著降低，脂肪颗粒明显减少，脂肪变性得到改善，表明苦参碱能够抑制 FFAs 诱导的 JNK、p38通路磷酸化。Q-PCR 结果表明苦参碱能促进肝实质细胞内 miR-122的表达，并降低脂肪化相关基因 Fas、Acc1的表达。结论苦参碱能显著改善高脂诱发的肝细胞脂肪样变性，并抑制 JNK、P38通路磷酸化，其机制可能同 miR-122通路相关。%Objective To investigate the effect of matrine on liver steatosis induced by free fatty acids (FFAs)in liver parenchymal cells.Methods Mouse primary hepatocytes were isolated by a two-step collagenase perfusion procedure and cultured in vitro.Cells were divided into three groups,including control group (CG),FFAs group (FG)and matrine treatment group (MG).The content of triglyceride was measured in vitro.Steatosis was analyzed by oil red O staining.The MAPK related signal pathway was assessed by western blot assay.Real-time polymerase chain reaction (PCR)was carried out to detect the expression of miR-122,which was closely related to fatty degeneration,and detect relevant target genes in liver parenchymal cells.Results Compared to that in FG,the content of triglyceride and fat particles in MG decreased significantly.MG could effectively reduce the level of the phosphorylation of JNK and p38 pathway,which was induced by FFAs.Q-PCR results showed that matrine up-regulated the expression of miR-122 while down-regulated the expression of Fas and Acc1 .Conclusion Matrine could significantly improve FFAs-induced steatosis and inhibit phosphorylation of JNK, p38 pathway,which might be through miR-122.