CAJ | 학술논문

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.
목적 탐토소벽감대C3H10T1/2지방세포자서적영향.방법 유도중배층적다능간세포주C3H10T1/2분화성지방세포,용소벽감처리,Western인적감측자서표지단백LC3Ⅱ/Ⅰ급자서저물P62적변화;용소벽감화용매체억제제록규공동처리,Western인적감측자서통량변화；투사전자현미경관찰소벽감처리후자서체적수량변화.결과 (1)소벽감처리후,Western인적현시LC3Ⅱ/Ⅰ강저,P62증고,병정시간여제량의뢰성(P＜0.05혹P＜0.01)；(2)록규처리후,Western인적현시LC3Ⅱ/Ⅰ증고,여록규조상비,록규화소벽감공동처리후,Western인적현시LC3Ⅱ/Ⅰ강저(P＜0.05)；(3)투사전자현미경현시소벽감처리후자서체수량명현감소.결론 소벽감능구억제C3H10T1/2지방세포적자서.
Objective To assess the effects ofberberine 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 LC3 Ⅱ/Ⅰ and autophagy substrate P62 was determined by Western blot ; after treating C3 H 10T1/2 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 LC3 Ⅱ / Ⅰin 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 LC3 Ⅱ/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.