小檗碱对C3H10T1/2脂肪细胞自噬的影响

Effects of berberine on autophagy in C3H10T1/2 adipocytes

目的 探讨小檗碱对C3H10T1/2脂肪细胞自噬的影响。方法 诱导中胚层的多能干细胞株C3H10T1/2分化成脂肪细胞,用小檗碱处理,Western印迹监测自噬标志蛋白LC3Ⅱ/Ⅰ及自噬底物P62的变化;用小檗碱和溶酶体抑制剂氯喹共同处理,Western印迹监测自噬通量变化;透射电子显微镜观察小檗碱处理后自噬体的数量变化。结果 （1）小檗碱处理后,Western印迹显示LC3Ⅱ/Ⅰ降低,P62增高,并呈时间与剂量依赖性（P〈0.05或P〈0.01）;（2）氯喹处理后,Western印迹显示LC3Ⅱ/Ⅰ增高,与氯喹组相比,氯喹和小檗碱共同处理后,Western印迹显示LC3Ⅱ/Ⅰ降低（P〈0.05）;（3）透射电子显微镜显示小檗碱处理后自噬体数量明显减少。结论 小檗碱能够抑制C3H10T1/2脂肪细胞的自噬。

Objective To assess the effects of berberine on autophagy in C3H10T1/2 adipocytes. Methods C3H10T1/2 cells, a pluripotent stem-cell line of mesodermal origin, were induced to differentiate into mature adipocytes, and then were treated with berberine, The expression of autophagy marker protein LC3Ⅱ/Ⅰ and autophagy substrate P62 was determined by Western blot; after treating C3H10T1/2 cells with berberine and lysosomal inhibitor, chloroquine, autophagy flux was assessed by Western blot. Autophagosome was observed by transmission electron microscopy after berberine treatment. Results （1） After berberine treatment, the expression of LC3Ⅱ/Ⅰ in C3H10T1/2 adipocytes was reduced and P62 was increased in a time-and dose-dependent manner（P〈0.05 or P〈0.01）; （2） Following treatment with chloroquine and berberine,the protein level of LC3Ⅱ/Ⅰ was decreased（P〈0.05）; （3） The number of autophagosome was decreased apparently after berberine treatment. Conclusions Berberine inhibits autophagy in C3H10T1/2 adipocytes.