益气活血方改善血糖波动型GK大鼠肾损伤的机制  

Mechanism of Yiqi Huoxue Recipe⁃mediated renal injury improvement in GK rats with glycemic fluctuation

作　　者：方文明 胡张捷 戴灵豪[1] 刘艺 王慧铭[1] 王辉[1] FANG Wen-ming;HU Zhang-jie;DAI Ling-hao;LIU Yi;WANG Hui-ming;WANG Hui(Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学,浙江杭州310053

出　　处：《中成药》2021年第7期1726-1733,共8页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金面上项目(81973767)。

摘　　要：目的探讨益气活血方是否通过TGF⁃β1/miR⁃21/PTEN/Akt信号通路改善血糖波动造成的糖尿病肾病损伤。方法将6只雄性Wistar大鼠作为正常组,通过上下午定时对GK大鼠注射葡萄糖和胰岛素制备血糖波动模型,将造模成功的GK大鼠随机分为模型组、缬沙坦组及益气活血方组。每周称量1次大鼠体质量,测量1次空腹血糖,给药8周后上代谢笼收集尿液,检测尿肌酐和24 h尿蛋白排泄率,颌下静脉采血,检测血清尿素氮、尿酸、总胆固醇、甘油三酯等指标。麻醉处死大鼠,取双肾称定质量,计算脏器指数。将单侧肾置于4%多聚甲醛保存,HE、PAS染色观察病理变化;另一单侧肾用于miR⁃21基因表达及TGF⁃β1(转化生长因子⁃β1)、PTEN(张力蛋白同源的磷酸酯酶基因)、P⁃Akt(磷酸化蛋白激酶B)蛋白表达检测。结果与模型组比较,益气活血方组的空腹血糖、尿肌酐、24 h尿蛋白排泄率、尿素氮、尿酸、总胆固醇、甘油三酯、肾脏指数、miR⁃21、TGF⁃β1及P⁃Akt表达均降低(P<0.05,P<0.01),PTEN表达升高(P<0.05),肾小球萎缩、基底膜增厚、细胞外基质积聚、系膜扩张及细胞形态改变均明显改善。结论益气活血方可能通过TGF⁃β1/miR⁃21/PTEN/Akt信号通路改善血糖波动造成的糖尿病肾病损伤。AIM To investigate whether Yiqi Huoxue Recipe can improve diabetic nephropathy(DN)damage due to glycemic fluctuation via TGF⁃β1/miR⁃21/PTEN/Akt signaling pathway.METHODS Six male Wistar rats were selected into the normal group.Glucose and insulin injections were applied to the 42 GK rats at regular intervals in the morning and afternoon to develop glycemic fluctuatng GK rat.models.The GK rats were randomly divided into three groups:model group,valsartan group and Yiqi Huoxue Recipe group.The rats had their weights and fasting blood glucose(FBG)measured once a week.After eight weeks of administration,the rats had the urine collected from the upper metabolic cage for determination of urinary creatinine and 24⁃hour urinary protein excretion rate,their blood samples collected from submaxillary vein for quantitative assessments of serum urea nitrogen,uric acid,total cholesterol and triglyceride.Finally,the rats killed under anesthesia were subjected to the kidney weight measurement and calculation of organ indices levels.To the bilateral kidneys,one unilateral kidney stored in 4%paraformaldehyde was conducted HE and PAS staining to observe the pathological changes,and another kidney underwent determinations of the expression of miR⁃21 gene and the protein expressions of TGF⁃β1,PTEN and P⁃Akt.RESULTS Compared with the model group,Yiqi Huoxue Recipe group shared significantly decreased levels of FBG,urinary creatinine,24⁃hour urinary protein excretion rate,urea nitrogen,uric acid,total cholesterol,triglyceride,renal index,miR⁃21 gene,TGF⁃β1 and P⁃Akt protein expression(P<0.05,P<0.01),significantly increased PTEN protein expression(P<0.05),and significantly improved glomerular atrophy,basement membrane thickening,extracellular matrix accumulation,mesangial expansion and cell morphological changes.CONCLUSION Yiqi Huoxue Recipe may improve the glycemic fluctuation⁃induced DN injury via TGF⁃β1/miR⁃21/PTEN/Akt signaling pathway.

关 键 词：益气活血方 糖尿病肾病 miR⁃21 PTEN P⁃Akt TGF⁃β1 

分 类 号：R285.5[医药卫生—中药学]