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目的观察五味子乙素(Sch B)对顺铂(CDDP)诱导的人类近端肾小管上皮细胞(HK-2)损伤中P21和Caspase-3表达的影响。方法取离体培养HK-2细胞,随机分为5组:对照组细胞未经任何处理,CDDP组加入50μmol/L CDDP处理24 h,Sch B预保护组(CDDP+Sch B):分别加入终浓度为5、10、20μmol/L Sch B预处理2 h后,其余操作同CDDP组。CCK-8试剂盒检测各组细胞活性;Annexin V-FITC/PI双标记流式细胞仪检测各组细胞凋亡率;倒置显微镜观察各组细胞形态;Western blotting检测各组细胞中P21和Caspase-3蛋白表达。结果与对照组相比,CDDP组细胞活性明显降低,细胞凋亡率、P21和Caspase-3蛋白表达量显著增加,细胞体积明显缩小,相互连接消失。Sch B预保护组较CDDP组细胞活性显著增加,细胞凋亡率、Caspase-3蛋白表达量显著减少,P21蛋白表达量显著增加,细胞形态损伤明显减轻,体积略微缩小,相互间连接断裂减少。结论 Sch B对CDDP诱导的HK-2细胞损伤有保护作用,其机制可能与其上调P21和下调Caspase-3的表达有关,且其保护作用呈剂量依赖性。
Objective To observe the effect of Sch B on the expression of P21 and Caspase-3 in human proximal tubular epithelial cells (HK-2) induced by cisplatin (CDDP). Methods HK-2 cells were isolated and cultured in vitro. The cells were randomly divided into 5 groups: control group, without any treatment, CDDP group treated with 50μmol / L CDDP for 24 hours, Sch B pre-protection group (CDDP + Sch B) The concentration of 5, 10, 20μmol / L Sch B pretreatment 2 h, the rest of the operation with the CDDP group. CCK-8 kit was used to detect the activity of each group; Annexin V-FITC / PI double-labeled flow cytometry was used to detect the apoptosis rate of each group; inverted microscope was used to observe the cell morphology; Western blotting was used to detect the expression of P21 and Caspase- 3 protein expression. Results Compared with the control group, the cell viability of CDDP group was significantly decreased. The apoptosis rate, the expression of P21 and Caspase-3 protein were significantly increased, the cell volume was significantly reduced, and disappeared. Compared with CDDP group, Sch B preincubation group significantly increased the cell viability, the apoptosis rate, Caspase-3 protein expression decreased significantly, P21 protein expression increased significantly, cell morphological damage was significantly reduced, the volume was slightly reduced, the connection between the fracture decreased. Conclusions Sch B can protect CDDP-induced HK-2 cell injury. The mechanism may be related to up-regulation of P21 and down-regulation of Caspase-3 expression and its protective effect in a dose-dependent manner.