摘要
目的探讨多肽Apelin在顺铂诱导的急性肾损伤中对肾小管上皮细胞线粒体的保护作用及相关机制。方法体外培养人近端肾小管上皮细胞,给予含顺铂(5μmol/L)和(或)Apelin-13(1μmol/L)的培养基孵育。设立3个实验组:正常对照组、顺铂组、顺铂+Apelin组。RT-PCR检测线粒体去乙酰化酶Sirt3 mRNA表达量;Western blot法检测线粒体乙酰化蛋白水平,以及线粒体分裂蛋白Drp1、融合蛋白OPA1表达量。选取8周龄雄性野生型小鼠,一次性腹腔注射顺铂(20mg/kg)诱导AKI。从顺铂注射的次日起,每日腹腔注射Apelin-13(0.1μmol/kg)或0.9%氯化钠注射液进行干预。设立3个实验组,即正常对照组、顺铂+生理盐水组、顺铂+Apelin组。透射电镜观察近端肾小管上皮线粒体形态,ELISA试剂盒检测血浆肌酐(Cr)、尿素氮(BUN)水平,以及尿液肾损伤分子-1(Kim-1)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)含量。结果细胞实验观察到,与正常对照组比较,顺铂组Sirt3 mRNA表达降低(P<0.05),线粒体乙酰化蛋白水平升高(P<0.05)。线粒体分裂蛋白Drp1表达增多(P<0.05),线粒体融合蛋白OPA1表达减少(P<0.05)。而顺铂+Apelin组,上述标志物表达量的变化受到显著抑制(P均<0.05)。动物实验显示,对照组小鼠肾脏线粒体结构完整。顺铂+生理盐水组,线粒体数量减少,形态扭曲伴空泡变。顺铂+Apelin组,线粒体的病变程度轻于顺铂+生理盐水组。与正常对照组比较,顺铂+生理盐水组小鼠血浆Cr、BUN升高,尿液Kim-1、NGAL含量明显增加(P均<0.05)。顺铂+Apelin组与顺铂+生理盐水组比较,上述指标的水平有所降低(P均<0.05)。结论Apelin通过增加线粒体去乙酰化酶Sirt3的表达,维持线粒体稳态,从而减轻顺铂诱导的急性肾损伤。
Objective To investigate the inhibitory effect of bioactive peptide apelin on tubular epithelial mitochondria in cisplatin-induced acute kidney injury(AKI).Methods Human proximal renal tubular epithelial cells were cultured in vitro and incubated with cisplatin(5μmol/L)and/or apelin-13(1μmol/L).Three experimental groups were set up:Control group,Cisplatin group,and Cisplatin+Apelin group.The mRNA expression of Sirt3 was detected by RT-PCR.The level of acetylation protein in mitochondria and expression of mitochondrial fission protein Drp1 and fusion protein OPA1 was detected by Western blot.AKI was induced by intraperitoneal injection of cisplatin(20mg/kg)for one time in 8-week-old male wild-type mice.Apelin-13(0.1μmol/kg)or saline were injected intraperitoneally every day from the next day of cisplatin injection.Three experimental groups were set up:Control group,Cisplatin+Saline group and Cisplatin+Apelin group.The proximal tubular mitochondria was observed by transmission electron microscopy.Plasma creatinine(Cr)and urea nitrogen,as well as,kidney injure molecule(Kim-1)and neutrophil gelatinase-associated lipocalin(NGAL)in urine,were detected by ELISA kit.Results Cell experiments showed that the expression of Sirt3 mRNA was significantly lower and the level of acetylation protein in mitochondria was higher in Cisplatin group than that in Control group(all P<0.05).The expression of mitochondrial fission protein Drp1 increased,meanwhile the expression of mitochondrial fusion protein OPA1 decreased in Cisplatin group than that in Control group(all P<0.05).The above changes were significantly inhibited in the Cisplatin+Apelin group(all P<0.05).Animal experiments showed that the structure of mitochondria in the kidneys of Control group was intact.In the Cisplatin+Saline group,the number of mitochondria decreased significantly,and mitochondrial morphology was distorted with vacuolation.In the Cisplatin+Apelin group,mitochondrial lesions were less severe than those in the Cisplatin+Saline group.Compared with Control group,the levels of plasma Cr,BUN,and urine Kim-1,NGAL were significantly higher in cisplatin+Saline group(all P<0.05).However,the above biomarkers in Cisplatin+Apelin group were significantly lower compared with Cisplatin+Saline group(all P<0.05).Conclusion Apelin protects the homeostasis of mitochondria by increasing the expression of deacetylase Sirt3,thereby alleviates cisplatin-induced acute kidney injury.
作者
王丽妍
关毅鸣
刁宗礼
刘文虎
Wang Liyan;Guan Yiming;Diao Zongli(Department of Nephrology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)
出处
《医学研究杂志》
2020年第5期36-41,共6页
Journal of Medical Research
基金
国家自然科学基金资助项目(81570660)
北京市医院管理局临床医学发展专项基金资助项目(ZYLX201824)
首都医科大学科研培育基金资助项目(自然类)(PYZ2018051)。
关键词
急性肾损伤
多肽
线粒体
去乙酰化酶
Acute kidney injury
Polypeptide
Mitochondrion
Deacetylase
