[关键词]
[摘要]
目的 观察五味子乙素(SchB)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,SchB预保护组(CDDP+SchB):分别加入终浓度为5、10、20 μmol/L SchB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;AnnexinV-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。SchB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 SchB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。
[Key word]
[Abstract]
Objective To explore the effect of schisandrin B (SchB) on P21 and Caspase-3 expression in cisplatin induced human proximal renal tubular epithelial (HK-2) cells injuried by CDDP in vitro. Methods HK-2 cells were randomly divided into five groups:control group (untreated), CDDP group (treated with 50 μmol/L CDDP for 24 h), SchB pretreat groups (CDDP+SchB, cells were pretreated with 5, 10, and 20 μmol/L SchB for 2 h, and the rest of the operation was the same as CDDP group). CCK-8 kit was used to detect the cell viability of five groups. Flow cytometry was used to detect the apoptotic rate of five groups. The morphology of cells was observed by inverted microscope. The protein expression of P21 and Caspase-3 was assessed by Western blotting assay. Results Compared with the control group, the cell viability decreased, and the apoptosis increased, the protein expression of P21 and Caspase-3 was up-regulated in HK-2 cells after treated with 50 μmol/L CDDP. Cell volume was reduced, and the mutual connection disappeared. Compared with the CDDP group, the cell viability increased and the apoptosis was decreased, the expression of Caspase-3 was down-regulated and the expression of P21 was up-regulated in HK-2 cells after pretreated with 5, 10, and 20 μmol/L SchB. Cell morphological damage lightened, the volume slightly decreased, and the mutual connection reduced. Conclusion SchB has a protective effect on HK-2 cells damage induced by cisplatin. The mechanism may be related to up-regulation of P21 and down-regulation of Caspase-3 expression, and its protective effect is dose dependent.
[中图分类号]
[基金项目]
天津市应用基础与前沿技术研究计划(14JCYBJC28200)