辣椒素对体外肝细胞脂质代谢的影响研究

Effects of capsaicin on lipid metabolism of liver cells in vitro

目的探讨辣椒素对脂肪变肝细胞内脂质沉积的影响及自噬表达情况,为脂肪性肝病防治的研究提供新的思路和靶点。方法用油酸(40μg/mL)诱导肝原代细胞LO2细胞株建立非酒精性脂肪性肝病(NAFLD)细胞模型。实验分为空白对照组、油酸组、辣椒素组[油酸+辣椒素(100μmol/L)]。油红O染色和三酰甘油试剂盒检测肝细胞内脂肪变程度;Western blot检测自噬体标记分子p62和LC3的表达水平并计算LC3-Ⅱ/LC3-Ⅰ比值。结果 40μg/mL油酸处理LO2细胞24h后,油红O染色观察,油酸组肝细胞中可见大小不等的橘红色脂滴,而空白对照组无明显橘红色脂滴形成,油酸在体外成功建立NAFLD模型。与油酸组相比,辣椒素组肝细胞内三酰甘油水平显著降低(P<0.05),脂滴明显减少,p62蛋白水平明显降低(P<0.05),而LC3-Ⅱ/LC3-Ⅰ比值明显上调(P<0.05)。结论在体外利用油酸诱导LO2细胞株建立NAFLD模型,辣椒素刺激NAFLD细胞,上调NAFLD细胞自噬水平,减少细胞内脂质沉积,但具体机制还需进一步研究。

Objective To observe the effect of capsaicin on lipid deposition in liver ceils and the expression of autophagy,to provide new ideas and targets for the study of fatty liver disease. Methods The model of nonalcoholic fatty liver disease （NAFLD） was established by inducing LGF cell line＇of liver primary ceils with oleic acid （40 μg/mL）. The experiment was divided into blank control group,oleic acid group, capsaicin group[oleic acid＋ capsaicin（100 μmol/L）]. The level of intracellular lipid was detected by oil red O staining and triglyceride （TG） kit. Western bolt was used to detect the expression level of autophagy marker molecule p62 and LC3,and to calculate the ratio of LC3- II/LC3- I . Results After treated with 40 μg/mL oleic acid for 24 h,the oil red O stai- ning showed that orange-red lipid droplets were found in the oleic acid group,but there was no obvious orange-red lipid droplet for- mation in the blank group,which suggesting that the NAFLD cell model was established successfully by using 40μg/mL oleic acid in vitro. Oil red O staining was observed,the ceils in the capsaicin group were significantly less than the oleic acid group, and the content of triglyceride in the liver cells was significantly lower than that in the oleic acid group（P〈0.05）. Western bolt test results showed that capsaicin group p62 protein levels were significantly lower compared with the oleic acid group（P〈0.05）, while the LC3-Ⅱ/LC3- Ⅰ ratio was higher than that of oleic acid group（P〈0.05）. Conclusion NAFLD model was established by inactiva- ting LO2 cell line with oleic acid in vitro. Capsaicin stimulated NAFLD cells, upregulated the autophagy level of NAFLD cells and reduced intracellular lipid deposition,but the specific mechanism need further study.