基于AMPK/mTOR自噬信号通路研究达格列净对糖尿病肾病模型大鼠肾损伤改善的作用机制  

Mechanism of dapagliflozin on the improvement of renal injury in a diabetic nephropathy rat model based on the AMPK/mTOR autophagy signaling pathway

作　　者：叶玉燕[1] 王鹏[2] 方霞 杨静[1] YE Yuyan;WANG Peng;FANG Xia;YANG Jing(Department of Nephrology,Jinhua People's Hospital,Jinhua,Zhejiang 321000,China;Department of Pharmacy,Jinhua People's Hospital,Jinhua,Zhejiang 321000,China)

机构地区：[1]浙江省金华市人民医院肾内科,金华321000 [2]浙江省金华市人民医院药学部,金华321000

出　　处：《浙江中西医结合杂志》2024年第10期905-909,914,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省公益类金华市科学技术研究计划项目(No.2021-4-067)。

摘　　要：目的研究达格列净对糖尿病肾病(diabetic nephropathy,DN)模型大鼠肾损伤的改善机制。方法40只清洁级Wistar雄性大鼠分为空白对照组(蒸馏水灌胃)、模型组(蒸馏水灌胃)、达格列净低剂量组[达格列净片1 mg/(kg·d)灌胃]、达格列净高剂量组[达格列净片10 mg/(kg·d)灌胃]各10只,干预12周。观察各组大鼠体质量、24 h尿量、24 h尿蛋白、随机血糖、血清尿素氮(BUN)、肌酐(Scr)的差异;苏木精-伊红染色观察各组大鼠肾脏组织病理变化;免疫组化检测各组大鼠肾组织腺苷酸活化蛋白激酶(AMPK)及哺乳动物雷帕霉素靶蛋白(mTOR)表达;实时荧光PCR(qRT-PCR)检测各组大鼠肾组织AMPK、mTOR mRNA表达。结果空白对照组、达格列净高剂量组、达格列净低剂量组、模型组大鼠体质量依次降低,24 h尿量、随机血糖、24 h尿蛋白、BUN、Scr依次升高,组间两两比较,差异均有统计学意义(P<0.01)。模型组、达格列净低剂量组、达格列净高剂量组大鼠肾脏组织病理变化比较,肾小球大小逐渐恢复,细胞排列依次紧密有序,肾小管上皮细胞空泡样变性逐渐减小,细胞排列整齐度依次增加,出血减少。空白对照组、达格列净高剂量组、达格列净低剂量组、模型组大鼠肾脏组织AMPK蛋白表达[(0.36±0.03)、(0.27±0.02)、(0.21±0.02)、(0.18±0.02)]依次降低(P<0.01),mTOR表达[(0.13±0.02)、(0.17±0.02)、(0.26±0.02)、(0.31±0.03)]依次升高(P<0.01),AMPK mRNA相对表达量[(1.00±0.08)、(0.77±0.07)、(0.52±0.05)、(0.26±0.03)]依次降低(P<0.01),mTOR mRNA相对表达量[(1.00±0.06)、(1.17±0.08)、(1.76±0.13)、(2.31±0.16)]依次升高(P<0.01)。结论达格列净可激活AMPK/mTOR自噬信号通路,保护肾小球滤过屏障,减少蛋白尿,延缓DN大鼠的疾病进程。Objective To study the mechanism of dapagliflozin in improving renal injury in diabetic nephropathy(DN)model rats.Methods Forty clean-grade male Wistar rats were divided into a blank control group(gavaged with distilled water),a model group(gavaged with distilled water),a low-dose group of dapagliflozin[gavaged with dapagliflozin 1 mg/(kg·d)],and a high-dose group of dapagliflozin[gavaged with dapagliflozin 10 mg/(kg·d)],with 10 rats in each group.After 12 weeks of intervention,body weight,24-hour urine volume,24-hour urine protein,random blood glucose,urea nitrogen(BUN),and creatinine(Scr)were measured and compared among the groups.Hematoxylin and eosin staining was used to observe the pathological changes of kidney tissue in each group.Immunohistochemistry was performed to detect the expression of adenosine monophosphate-activated protein kinase(AMPK)and mammalian target of rapamycin(mTOR)proteins in rat renal tissue.Real-time fluorescence quantitative PCR was used to detect the expression of AMPK and mTOR mRNAs in rat renal tissue.Results Among the blank control,high-dose dapagliflozin,low-dose dapagliflozin,and model groups,the body weight decreased sequentially,while 24-hour urine volume,random blood glucose,24-hour urine protein,BUN,and Scr increased sequentially,with significant differences between groups(P<0.01).Pathological examination revealed that among the model,low-dose dapagliflozin,and high-dose dapagliflozin groups,the glomerular size gradually returned to normal,the cell arrangement was tightly and orderly arranged,the vacuolar degeneration of renal tubular epithelial cells gradually decreased,and the cell arrangement became more organized with reduced bleeding.Among the blank control,high-dose dapagliflozin,low-dose dapagliflozin,and model groups,AMPK protein expression[(0.36±0.03),(0.27±0.02),(0.21±0.02),(0.18±0.02)]decreased sequentially(P<0.01),while the mTOR expression[(0.13±0.02),(0.17±0.02),(0.26±0.02),(0.31±0.03)]increased sequentially(P<0.01).The relative expression level of AM

关 键 词：大鼠 糖尿病肾病 达格列净 腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白自噬信号通路 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]