达格列净通过mTOR自噬信号通路改善糖尿病肾病大鼠足细胞损伤  

Dapagliflozin ameliorates podocyte injury in diabetic nephropathy rats through mTOR autophagy signaling pathway

作　　者：万倩 周华 惠辉[1] 王脉桃 张亚萍[1] 宋卫红[1] WAN Qian;ZHOU Hua;HUI Hui;WANG Maitao;ZHANG Yaping;SONG Weihong(Department of Endocrine,First People’s Hospital of Chenzhou City,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院内分泌科,郴州423000

出　　处：《山西医科大学学报》2022年第11期1439-1445,共7页Journal of Shanxi Medical University

基　　金：郴州市技术创新引导项目(lcyl2021070)。

摘　　要：目的探讨达格列净对糖尿病肾病(diabetic nephropathy,DN)大鼠足细胞损伤的影响及相关分子机制。方法46只Wistar雄性大鼠以随机数字表法分为对照组(NC组,n=10)和造模组(n=36)。成功构建DN模型的30只造模组大鼠随机分为模型组(DN组)、达格列净组(Dapa组)、达格列净+雷帕霉素组(Dapa+Rapa组),每组10只。各组大鼠均给药干预12周,1次/d。采用血糖仪检测大鼠空腹血糖(FBG)。全自动生化分析仪测定大鼠血肌酐(Scr)、尿素氮(BUN)、24 h尿蛋白(UP)。HE染色观察大鼠肾组织病理形态学。透射电子显微镜观察大鼠肾脏足细胞变化。Western blot法检测足细胞标志蛋白(nephrin、podocin)、自噬蛋白(Beclin-1、LC3Ⅱ、P62)及mTOR信号通路相关蛋白(mTOR、p-mTOR)表达。结果与NC组比较,DN组大鼠肾小球体积增大,肾小管上皮细胞空泡变性,存在大量炎性细胞浸润;足细胞足突融合,排列紊乱,可见明显线粒体嵴断裂或空泡化。与DN组比较,Dapa组和Dapa+Rapa组大鼠肾脏病理损伤明显减轻;足细胞可见部分足突融合,排列较整齐,整体情况明显改善。与Dapa组比较,Dapa+Rapa组大鼠肾脏病理损伤和足细胞整体情况显著改善。与NC组比较,DN组大鼠血清FBG、SCr、BUN、24 h UP水平均升高(P<0.05);肾组织nephrin、podocin、Beclin-1、LC3Ⅱ蛋白表达水平均下降,P62蛋白表达水平、p-mTOR/mTOR比值均升高(P<0.05)。与DN组比较,Dapa组和Dapa+Rapa组大鼠血清FBG、SCr、BUN、24 h UP水平均降低(P<0.05),肾组织nephrin、podocin、Beclin-1、LC3Ⅱ蛋白表达水平均升高(P<0.05),P62蛋白表达水平、p-mTOR/mTOR比值均降低(P<0.05)。与Dapa组比较,Dapa+Rapa组大鼠血清FBG、SCr、BUN、24 h UP水平均降低(P<0.05),肾组织nephrin、podocin、Beclin-1、LC3Ⅱ蛋白表达水平均升高(P<0.05),P62蛋白表达水平、p-mTOR/mTOR比值均降低(P<0.05)。结论达格列净通过调节足细胞自噬减轻DN足细胞损伤,进而改善DN,其作用机制�Objective To investigate the effect of dapagliflozin on podocyte injury in diabetic nephropathy(DN)rats and its molecular mechanism.Methods Forty-six Wistar male rats were randomly divided into control group(NC group,n=10)and model group(n=36).Thirty DN model rats were randomly divided into model group(DN group),dapagliflozin group(Dapa group),dapagliflozin+rapamycin group(Dapa+Rapa group),with 10 rats in each group.The rats in each group were given the corresponding drug once a day for 12 weeks.Fasting blood glucose(FBG)was measured by blood glucose meter.Serum creatinine(Scr),urea nitrogen(BUN),24-hour urine protein(UP)were measured by automatic biochemical analyzer.The renal histopathology was observed by HE staining.The changes of rat renal podocytes were observed under transmission electron microscope.The expression levels of podocyte marker proteins(nephrin,podocin),autophagic proteins(Beclin-1,LC3Ⅱ,P62)and mTOR signaling pathway-related proteins(mTOR,p-mTOR)were detected by Western blot.Results Compared with NC group,the glomerular volume was increased in DN group,the renal tubular epithelial cells were vacuolated and denatured,and there were a large number of inflammatory cells infiltration:the podocyte foot process fusion and disordered arrangement,with obvious mitochondrial cristae rupture or vacuolization.Compared with DN group,the renal pathological damages were significantly alleviated in Dapa group and Dapa+Rapa group:partial foot process fusion and orderly arrangement of the podocytes,and the overall situation was significantly improved.Compared with Dapa group,the renal pathological damage and the overall situation of podocytes were significantly improved in Dapa+Rapa group.Compared with NC group,the serum levels of FBG,Scr,BUN and 24 h UP were increased in DN group(P<0.05),the expression levels of nephrin,podocin,Beclin-1 and LC3Ⅱin renal tissue were decreased(P<0.05),while the expression level of P62 and the ratio of p-mTOR/mTOR in renal tissue were increased(P<0.05).Compared with DN group,t

关 键 词：达格列净 糖尿病肾病 足细胞 自噬 MTOR信号通路 

分 类 号：R587.2[医药卫生—内分泌]