苦参碱对顺铂诱导的肾小管上皮细胞损伤的保护作用

The protective effect of matrine on the cisplatin-induced rat renal tubular epithelial cell injury

目的:研究苦参碱对顺铂所致大鼠肾小管上皮细胞NRK-52E损伤的保护作用。方法:将培养的肾小管上皮细胞分为3组:对照组(不干预)、顺铂组(加入50μmol/L顺铂处理24h)、顺铂+苦参碱组(加入顺铂50μmol/L和苦参碱0.5mg/mL处理24h)。显微镜下观察NRK-52E细胞形态;采用CCK-8法和乳酸脱氢酶(LDH)细胞毒性实验检测细胞生长及损伤情况;实时荧光定量PCR(qPCR)法和Western blotting法分别检测NRK-52E细胞自噬相关标志蛋白微管相关蛋白1轻链3(LC3)基因和蛋白表达水平。结果:与对照组比较,顺铂诱导NRK-52E细胞出现形态学改变,细胞存活力下降,细胞损伤增加,LC3 mRNA水平及LC3-Ⅱ/LC3-Ⅰ比值升高(P<0.05)。与顺铂组比较,苦参碱干预能够改善顺铂诱导的细胞形态变化,提高细胞存活力,降低细胞损伤,并进一步升高LC3mRNA水平及LC3-Ⅱ/LC3-Ⅰ比值(P<0.05)。结论:苦参碱能够改善顺铂诱导的肾小管上皮细胞毒性损伤,其可能通过增强细胞自噬水平来发挥肾保护作用。

Objective: To study the effect of matrine on cisplatin-induced damage in rat renal tubular epithelial cells NRK-52E. Methods: NRK-52E cells were divided into three groups:control group (non-treated cells),cisplatin group (cells were cultured with 50 μmol/L cisplatin for 24 h),and cisplatin+matrine group (cells were cultured with 50 μmol/L cisplatin and 0.5 mg/mL matrine for 24 h).The morphologic changes of NRK-52E cells were observed under an inverted microscope.CCK-8 assay and lactate dehydrogenase (LDH) cytotoxicity assay were used to detect the proliferation and survival rate of NRK-52E cells.The fluorescence quantitative PCR (qPCR) and western blotting were used to detect the mRNA and protein expressions of microtubule associated protein 1 light chain 3 (LC3),respectively. Results: Compared with the control group,the morphology of cells obviously changed,the cell viability was decreased,the cell death,the LC3 mRNA level and the ratio of LC3-Ⅱ/LC3-Ⅰ were increased after treatment with cisplatin ( P <0.05).The cytotoxicity of cisplatin was inhibited by matrine ( P <0.05).The autophagy-related marker LC3 mRNA level and LC3-Ⅱ/LC3-Ⅰ ratio were markedly higher in the cisplatin+matrine group than those in the cisplatin group ( P <0.05). Conclusion: Matrine could protect against cisplatin-induced injury in renal tubular epithelial cells,and the mechanism might be related to the enhancement of autophagy.