摘要
目的:探讨miR-101靶向雷帕霉素靶蛋白(mTOR)信号通路调控足细胞自噬在小鼠糖尿病肾病发病中的作用。方法:将成年雄性C57BL/6小鼠分为对照组、模型组和雷帕霉素组,每组10只。模型组和雷帕霉素组小鼠连续腹腔注射链脲佐菌素(55 mg·kg^(-1))5 d,对照组腹腔注射等量生理盐水。腹腔注射链脲佐菌素后1周,雷帕霉素组小鼠腹腔注射雷帕霉素(2 mg·kg^(-1)),对照组和模型组小鼠腹腔注射等量生理盐水,隔天1次,连续12周后对小鼠肾脏组织行PAS染色,用透射电镜观察足细胞损伤及自噬情况,同时检测肾组织中miR-101、Nephrin、LC3Ⅰ、LC3Ⅱ、mTOR和AMPK mRNA的表达。结果:对照组小鼠肾脏和足细胞足突正常,模型组小鼠肾脏局部可见球囊融合;与对照组相比模型组小鼠肾脏系膜基质明显增多,基底膜明显增厚,足突明显增宽、融合,自噬体明显减少;与模型组相比,雷帕霉素组小鼠肾脏系膜基质明显减少,基底膜增厚和足突融合明显减轻,自噬体明显增加。与对照组相比,模型组和雷帕霉素组小鼠肾脏系膜基质面积增大、mTOR mRNA表达水平明显升高,Nephrin、LC3Ⅰ、LC3Ⅱ、miR-101和AMPK mRNA表达水平明显降低(P<0.05);与模型组相比,雷帕霉素组小鼠mTOR mRNA表达水平明显降低,Nephrin、LC3Ⅰ、LC3Ⅱ、miR-101和AMPK mRNA表达水平明显升高(P<0.05)。结论:miR-101通过靶向调控mTOR信号通路促进糖尿病肾病小鼠足细胞自噬,为糖尿病肾病的治疗提供了新方向。
Objective:To investigate the role of miR-101 targeting mammalian target of rapamycin(mTOR)signaling pathway in regulating podocyte autophagy in the pathogenesis of diabetic nephropathy in mice.Methods:The adult male C57BL/6 mice were divided into control group,model group and rapamycin group with 10 mice in each group.Mice in the model group and rapamycin group were intraperitoneally injected with streptozotocin(55 mg·kg^(-1))for 5 days,and the control group was intraperitoneally injected with the same amount of normal saline.One week after intraperitoneal injection of streptozotocin,mice in the rapamycin group were intraperitoneally injected with rapamycin(2 mg·kg^(-1)),and mice in the control and model groups were intraperitoneally injected with the same amount of normal saline,once every other day.After 12 weeks,the kidney tissue was stained with PAS,the podocyte damage and autophagy were observed by transmission electron microscope.At the same time,the expression of miR-101,Nephrin,LC3Ⅰ,LC3Ⅱ,mTOR and AMPK mRNA in the kidney tissue was detected.Results:The kidneys and podocyte foot processes of the mice in the control group were normal,and the kidneys of the model group mice were locally fused with balloons.Compared with the control group,the mesangial matrix in the model group was significantly increased,the basement membrane significantly thickened,and the foot processes were significantly widened and fused.Compared with the model group,the mesangial matrix of the kidney of mice in the rapamycin group was significantly reduced,the basement membrane thickening and foot process fusion significantly reduced,and the autophagosomes were significantly increased.Compared with the control group,the mesangial matrix area and level of mTOR mRNA in the model group and the rapamycin group were significantly increased,and the levels of Nephrin,LC3Ⅰ,LC3Ⅱ,miR-101 and AMPK mRNA were significantly reduced(P<0.05).Compared with the model group,the mesangial matrix area and mTOR mRNA level of mice in the rapamycin gr
作者
胡玲
周乐汀
王思思
王凉
孙铸兴
HU Ling;ZHOU Leting;WANG Sisi;WANG Liang;SUN Zhuxing(Department of Nephrology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023,China)
出处
《东南大学学报（医学版）》
CAS
2021年第2期214-219,共6页
Journal of Southeast University(Medical Science Edition)
基金
无锡市卫生计生委科研项目(Q201761)。
