苦参碱对阿霉素损伤足细胞的干预作用及mTOR的表达

摘要

目的探讨苦参碱（MAT）对阿霉素（ADR）损伤足细胞的干预作用及哺乳动物雷帕霉素靶蛋白（mTOR）在此过程中的表达。方法用1μmol/L ADR制造足细胞损伤模型，分别以10、20、40μg/ml MAT干预24 h，检测足细胞活力、足细胞凋亡比例、足细胞中间丝蛋白（Desmin）和mTOR表达，分析不同浓度MAT干预ADR损伤足细胞的效果。结果与对照组相比，ADR损伤模型的足细胞活性下降、足细胞凋亡比例增加、损伤标志分子Desmin表达增多，差异均有统计学意义（P<0.01）；给予10~40μg/ml MAT干预后，相对于损伤模型组，足细胞活性增加、凋亡减少、Desmin表达下降，差异均有统计学意义（P<0.05）。足细胞受损后，mTOR和磷酸化mTOR表达均下降，差异有统计学意义（P<0.05）；给予10~40μg/ml MAT干预，mTOR、p-mROR表达较损伤模型组增加，差异有统计学意义（P<0.05）。足细胞受损后，mTOR下游靶蛋白S6K1和4EBP1 mRNA表达下降，但差异未达到统计学意义；予10~40μg/ml MAT干预后，S6K1和4EBP1 mRNA表达回升，差异有统计学意义（P<0.05）。结论10~40μg/ml MAT对ADR诱导足细胞损伤有保护作用，其保护机制可能与mTOR信号通路有关。%Objectives To investigate the effect of matrine (MAT) on adriamycin (ADR) induced podocyte injury in vitro and explore the function of the mammalian target of rapamycin (mTOR) protein signaling pathway during the intervention. Methods ADR (1μmol/L) was used to induce the model of podocyte injury and then the podocytes were intervened by 10, 20 and 40μg/ml MAT for 24 hours. The viability and apoptosis rate of podocytes, mRNA and protein expressions of desmin and mTOR were detected. The effect of different concentrations of MAT on ADR-induced podocytes was analyzed. Results Com-pared with the control group, declined viability of podocytes (P<0.001), increased percentage of apoptotic podocytes (P<0.001), and increased expression of damage marker desmin (P=0.002) were observed in ADR group. In ADR group, after intervention with MAT of 10-40μg/ml, increased viability of podocytes (P<0.05), decreased percentage of apoptotic podocytes (P<0.05), and decreased expression of damage marker Desmin were found (P<0.05). The protein expression of mTOR and phosphorylation state of mTOR (p-mTOR) decreased in ADR induced-podocytes (P<0.05), and after intervention with MAT of 10-40μg/ml, the expression of mTOR and p-mTOR increased (P<0.05). The expression of mTOR downstream target protein s6k1 and 4EBP1 mRNA decreased in ADR group (P=0.071), while increased after intervention with MAT of 10-40μg/ml (P<0.05). Conclusions 10-40μg/ml MAT have protective effects on ADR-induced podocytes in vitro. The protection mechanism may be related to mTOR signaling pathway.