蟾蜍灵对体外庆大霉素诱导的大鼠肾小管上皮细胞自噬的影响  

Effects of bufalin on autophagy induced by gentamicin in rat renal tubular epithelial cells in vitro

作　　者：丁玲[1] 施会敏 张爱青[1] 甘卫华[1] 

机构地区：[1]南京医科大学第二附属医院儿科,江苏南京210003

出　　处：《南京医科大学学报（自然科学版）》2017年第9期1120-1123,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学科技发展基金面上项目(2014NJMU135);江苏省高校自然科学研究面上项目(15KJD320005)

摘　　要：目的:观察蟾蜍灵对体外庆大霉素诱导的大鼠肾小管上皮细胞自噬的影响。方法:体外培养大鼠肾小管上皮细胞,分为正常对照组、庆大霉素组(2 mg/m L)、蟾蜍灵组(1×10^(-8)mol/L)、庆大霉素+蟾蜍灵组(庆大霉素2 mg/mL+蟾蜍灵1×10^(-8) mol/L)。应用透射电镜观察各组大鼠肾小管上皮细胞的自噬水平;Western blot检测庆大霉素及蟾蜍灵对肾小管上皮细胞自噬标志蛋白LC3Ⅱ、P62及肾损伤分子1(kindey injury molecule-1,KIM1)表达的影响。结果:透射电镜下观察:与对照组相比,庆大霉素组自噬相关形态指标自噬体、自噬泡等数量明显增加;蟾蜍灵干预后,细胞自噬体数量较庆大霉素组明显减少。Western blot结果显示,与对照组相比,庆大霉素组自噬标志蛋白LC3Ⅱ、P62、KIM1的蛋白相对表达量均升高(P<0.05),蟾蜍灵干预后,LC3Ⅱ、P62及KIM1蛋白相对表达量较庆大霉素组下降(P<0.05)。结论:庆大霉素可促进大鼠肾小管上皮细胞发生自噬、提高KIM1蛋白的表达;蟾蜍灵可部分缓解庆大霉素诱导的肾小管上皮细胞自噬的增加及KIM1蛋白表达量的提高,提示这可能是其发挥肾脏保护作用的机制之一。Objective：To analyze the effects of bufalin on autophagy induced by gentamicin （GM） in rat renal tubular epithelial cells （NRK52e） in vitro.Methods：NRK52e cells were cultured in vitro and divided into 4 groups： the control group,the GM group （2 mg/m L）,the bufalin group （1×10-8mol/L） and the GM＋bufalin group （GM 2 mg/m L＋bufalin 1×10-8mol/L）.The autophagy of NRK52 e cells was observed by transmission electron microscopy.Effects of GM and bufalin on the expression of autophagy-related protein LC3 Ⅱ and P62,and kidney injury molecule-1 （KIM1） protein were examined by Western blot.Results：Transmission electron microscopy showed that the number of autophagosomes in the GM group was significantly increased compared with the control group,and the number of autophagosomes of the bufalin group was less than that in the GM group.Western blot showed that the relative expressions of LC3Ⅱ,P62 and KIM1 protein were significantly increased in the GM group compared with the control group （all P〈0.05）,however,the relative expressions of LC3Ⅱ,P62 and KIM1 protein were decreased in the bufalin group than those in the GM group （P〈0.05）.Conclusion：Gentamicin could activate the autophagy of NRK52e cells and up-regulate the expression of KIM1 protein,while bufalin could partially inhibit GM-induced NRK52e cell＇s autophagy and the up-regulated expression of KIM1 protein.It may be the mechanisms by which bufalin acts the renoprotection.

关 键 词：蟾蜍灵 庆大霉素 肾小管上皮细胞 自噬 肾损伤分子1 

分 类 号：R729[医药卫生—儿科]