摘要
目的探讨达格列净通过激活腺苷酸活化蛋白激酶(AMPK)途径增强足细胞自噬。方法20只4~5周龄SD雄性大鼠随机选取6只为空白对照组(Con),其余有12只成功建模T2DM大鼠,T2DM大鼠随机分为T2DM组及达格列净组(Dap),每组各6只。Dap组予达格列净灌胃治疗4周后,收集标本进行检测,电镜观察肾脏结构变化,免疫荧光、Western blot检测相关蛋白表达水平。结果电镜下Dap组肾小球区病变轻于T2DM组。Dap组肾重/体重、FPG、FIns、24 h尿总蛋白、24 hUAlb低于T2DM组(P<0.05)。与Con组比较,T2DM组自噬标志物微管相关蛋白轻链3(LC3-Ⅱ/LC3-I)比值、自噬同源蛋白(Beclin-1)、足细胞特异标记蛋白(NPHS2)降低(P<0.05),Dap组p-AMPK/AMPK比值、Beclin-1蛋白、LC3-Ⅱ/LC3-I比值升高(P<0.05),NPHS2降低(P<0.05)。与T2DM组比较,Dap组p-AMPK/AMPK比值、Beclin-1、NPHS2、LC3-Ⅱ/LC3-Ⅰ比值升高(P<0.05)。结论达格列净可通过激活AMPK途径增强T2DM大鼠足细胞自噬发挥肾脏保护作用。
Objective To investigate whether Dapagliflozin can induce podocyte autophagy by activating adenosine monophosphate-activited protein kinase(AMPK)pathway.Methods A total of 20male diabetic rats 4~5 week old were randomly divided into three groups:diabetes group(T2DM,n=6),Dapagliflozin group(Dap,n=6),and control group(Con,n=6).Dap group were treated with Dapagliflozin for 4 weeks,and the corresponding specimens were collected and examined after treatment.The changes of renal structure were evaluatedby electron microscopy,and the expression levels of related proteins were detected by immunofluorescence and western blot.Results Under electron microscope,the glomerular lesion was slighterin the Dapagliflozin group than in T2DM group.The ratio of kidney weight to body weight,serum glucose,serum insulin,24-hour urinary total protein and 24-hour urinary albumin were lower in Dap group than in T2DM group(P<0.05).Compared with Con group,autophagy marker microtubuleassociated protein light chain 3(LC3-Ⅱ/LC3-I)protein ratio and autophagy homologous protein(Beclin-1)protein,podocyte marker(NPHS2)protein were decreased in T2DM group(P<0.05),while p-AMPK/AMPK ratio,Beclin-1 protein and LC3-Ⅱ/LC3-Ⅰratio were increased and NPHS2 protein was decreased in Dap group(P<0.05).Compared with T2DM group,p-AMPK/AMPK ratio,Beclin-1 protein,NPHS2protein and LC3-Ⅱ/LC3-Ⅰratio were increased in Dap group(P<0.05).Conclusion Dapagliflozin can enhance the autophagy of podocytesby activating AMPK pathway and play a protective role in kidney in T2DM rats.
作者
熊哲学
李凝旭
唐明娟
XIONG Zhexue;LI Ningxu;TANG Mingjuan(Deparlment of Nephrology,Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430000,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2023年第1期53-59,共7页
Chinese Journal of Diabetes
基金
国家重点研发计划基金资助项目(SQ2018YFC200194-02)。
