雷帕霉素对高糖状态下肾小球系膜细胞自噬的影响  

Effect of rapamycin on autophagy of mesangial cells under high glucose condition

作　　者：付雅 郝艳霞[1] 贾奇凤 陈洋洋 李利 王燕[1] 薛月梅 周婷 张亮[1] Fu Ya;Hao Yanxia;Jia Qifeng;Chen Yangyang;Li Li;Wang Yan;Xue Yuemei;Zhou Ting;Zhang Liang(Department of Nephrology,Ordos Central Hospital,Inner Mongolia Autonomous Region,Ordos 017000,China)

机构地区：[1]内蒙古自治区鄂尔多斯市中心医院肾脏内科,鄂尔多斯017000

出　　处：《国际泌尿系统杂志》2024年第4期692-696,共5页International Journal of Urology and Nephrology

基　　金：内蒙古自治区自然科学基金项目(2020MS08089)。

摘　　要：目的探讨雷帕霉素对高糖状态下肾小球系膜细胞自噬的影响。方法将人肾小球系膜细胞(HMC)在恒温培养箱中培养,将细胞分为HMC组、低糖组、高糖组、甘露醇等渗组和雷帕霉素组。采用GFP-RFP-LC3标记各组细胞的自噬流;采用Western blot检测各组细胞的自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ、P62的表达情况;采用ELISA法测定各组细胞的细胞Ⅳ型胶原纤维(ColⅣ)、透明质酸(HA)、层粘连蛋白(LA)的含量。结果高糖组的自噬溶酶体最少,HMC组、低糖组、甘露醇等渗组、雷帕霉素组的自噬溶酶体均较高糖组多,其中雷帕霉素组的自噬溶酶体最多。与HMC组、低糖组、甘露醇等渗组相比,高糖组的LC3-Ⅱ表达明显减少,LC3-Ⅰ表达明显增多,P62明显增多(均P<0.05)。雷帕霉素组的LC3-Ⅱ表达较高糖组明显增多,而LC3-Ⅰ及P62表达明显减少(均P<0.05)。高糖组的ColⅣ、HA、LA含量均较HMC组、低糖组和甘露醇等渗组明显升高,差异均有统计学意义(均P<0.001)。雷帕霉素组的ColⅣ、HA、LA的含量均较高糖组明显下降,差异均有统计学意义(均P<0.05)。结论高糖状态可以抑制HMC自噬活性,增加细胞外基质的聚集,促进糖尿病肾病进展,而雷帕霉素可诱导细胞自噬的发生,减少肾小球系膜细胞系膜基质增生,进而抑制糖尿病肾病进展。Objective To investigate the effect of rapamycin on autophagy of mesangial cells in high glucose state.Methods Human mesangial cells(HMC)were cultured in a constant temperature incubater and divided into HMC group,low glycemic group,high glycemic group,mannitol isotonic group and rapamycin group in constant temperature incubator.The autophagy flow of cells in each group was marked by GFP-RFP-LC3.Western blot was used to detect the expression of autophagy related proteins LC3-Ⅱ/LC3-Ⅰand P62 in each group.The contents of cell typeⅣcollagen fiber(ColⅣ),hyaluronic acid(HA)and laminin(LA)were determined by ELISA.Results Compared with HMC group,low glycemic group and mannitol isotonic group,LC3-Ⅱexpression was significantly decreased,LC3-Ⅰexpression was significantly increased and P62 expression was significantly increased in high glycemic group.The expression of LC3-Ⅱin rapamycin group increased significantly,while the expression of LC3-Ⅰand P62 decreased significantly in high glycemic group,with statistical significance(all P<0.05).The contents of ColⅣ,HA and LA in high glycemic group were significantly higher than those in HMC group(all P<0.001).The contents of ColⅣ,HA and LA in rapamycin group were significantly decreased in high glycemic group,with statistical significance(all P<0.05).Conclusions High glucose status can inhibit the autophagy activity of human mesangial cells,increase the aggregation of extracellular matrix,and promote the progression of diabetic nephropathy,while rapamycin can induce the occurrence of autophagy,reduce the proliferation of mesangial matrix of glomerular mesangial cells,and then inhibit the progression of diabetic nephropathy.

关 键 词：肾小球系膜细胞 自噬 雷帕霉素 高糖 

分 类 号：R692[医药卫生—泌尿科学]