羟苯磺酸钙对顺铂诱导的HK-2细胞凋亡的抑制作用及其机制  

The inhibitory effect of calcium dobesilate on cisplatin induced HK-2 cell apoptosis and its mechanism

作　　者：任春霞 朱李飞 余自成 REN Chun-xia;ZHU Li-fei;YU Zi-cheng(Department of Pharmacy,Yangpu Hospital,Tongji University,Shanghai 200090,China)

机构地区：[1]同济大学附属杨浦医院药学部,上海200090

出　　处：《药学学报》2020年第8期1830-1835,共6页Acta Pharmaceutica Sinica

基　　金：上海市卫生和计划生育委员会科研课题(20184Y0227);上海市科学技术委员会科研计划项目(19ZR1450600);上海市临床药学重点专科建设项目支持课题。

摘　　要：研究羟苯磺酸钙(calcium dobesilate,CaD)对顺铂(cisplatin,CP)致人肾近端小管上皮细胞(HK-2)凋亡的影响并探讨其可能的作用机制。利用顺铂诱导建立HK-2凋亡模型,采用CCK-8法检测CaD对HK-2增殖的影响;用流式细胞术检测细胞凋亡;用活性氧(reactive oxygen species,ROS)检测观察氧化应激水平;用JC-1法测线粒体膜电位;利用Western blot法检测顺铂诱导HK-2的p53、caspase-3、bcl-2和bax蛋白表达水平;用ELISA法检测肾损伤因子1(kidney injury molecule-1,KIM-1)、中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase associated lipocalin,NGAL)的表达。研究显示,CaD能够降低顺铂引起的氧化应激水平,抑制细胞凋亡。顺铂可上调p53、caspase-3、bax、KIM-1及NGAL的蛋白表达,降低bcl-2的表达。使用CaD可明显减少KIM-1、NGAL、p53、caspase-3和bax蛋白水平,促使bcl-2表达水平升高。此研究结果表明,CaD可通过抑制HK-2凋亡减轻顺铂诱导的肾小管上皮细胞损伤,其作用机制可能与调控bax/bcl-2/caspase-3凋亡信号通路有关。We observed the effect of calcium dobesilate(CaD)on apoptosis induced by cisplatin in human proximal tubular epithelial cells(HK-2)and explored the possible mechanism.Based on HK-2 cells apoptosis model induced by cisplatin,CCK-8 method was used to detect the effect of CaD on the proliferation of HK-2.Apoptosis was detected by flow cytometry.Reactive oxygen species(ROS)assay was used to evaluate the level of oxidative stress.The mitochondrial membrane potential was measured by JC-1 method.The expression levels of p53,caspase-3,bcl-2 and bax in cisplatin-induced HK-2 were detected by Western blot.The expression of renal injury factor 1(KIM-1)and neutrophil gelatin-related apolipoprotein(NGAL),markers of acute kidney injury,were detected by ELISA.The results showed that CaD could reduce the oxidative stress level induced by cisplatin and inhibit apoptosis in renal tubular epithelial cells.Cisplatin can up-regulate the protein expressions of p53,caspase-3,bax,KIM-1 and NGAL,and reduce the expression of bcl-2.After using CaD,the protein levels of KIM-1,NGAL,p53,caspase-3 and bax were significantly reduced,while the levels of bcl-2 were increased.This study has shown that CaD can alleviate cisplatin-induced HK-2 injury and inhibit HK-2 apoptosis,which may be related to the regulation of bax/bcl-2/caspase-3 apoptosis signaling pathway.

关 键 词：羟苯磺酸钙 顺铂 人肾近端小管上皮细胞 细胞凋亡 氧化应激 

分 类 号：R965[医药卫生—药理学]