芪归药对基于VEGF/PI3K/Akt信号通路干预糖尿病肾病大鼠肾小管间质纤维化的分子机制  

Molecular Mechanism of Huangqi(Astragali Radix)-Danggui(Angelicae Sinensis Radix)Medicinal Pairs on Tubulointerstitial Fibrosis in Rats with Diabetic Nephropathy Based on VEGF/PI3K/Akt Signaling Pathway

作　　者：沈梦琪 赵泉霖[2] SHEN Mengqi;ZHAO Quanlin(Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong,China)

机构地区：[1]山东中医药大学,山东济南250355 [2]山东中医药大学附属医院,山东济南250014

出　　处：《辽宁中医药大学学报》2023年第11期59-63,F0003,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：山东省自然科学基金资助项目(H2708)。

摘　　要：目的探讨黄芪配伍当归药对通过VEGF/PI3K/Akt通路干预糖尿病肾病(diabetic nephropathy,DN)对大鼠肾小管间质纤维化水平的影响,探究黄芪当归药对对于DN的作用分子机制及疗效的影响。方法采用链脲佐菌素(streptozocin,STZ)30 mg/kg腹腔注射进行DN大鼠模型的构建,分为模型组、芪归药对组(黄芪∶当归=1∶1、5.40 g·kg^(-1)·d^(-1))、达格列净组(0.45 mg·kg^(-1)·d^(-1)),每组各5只,以同周龄SD大鼠5只作为正常对照组。此外,给药期间分别测定各组大鼠体质量、尿量、尿糖、尿酮、空腹血糖INS,给药前和最后一次给药后进行HbA1c检查。8周后处死,各组均选择3只分离肾脏,按病理组织学资料制作肾组织石蜡切片,用于进行肾组织病理学、超微结构形态学的观察,RT-PCR法检测DN大鼠肾脏组织VEGF、PI3K及Akt的miRNA表达量,免疫组化法检测VEGF、PI3K及Akt的蛋白表达情况,以探究芪归药干预VEGF/PI3K/Akt通路对DN大鼠肾小管间质纤维化进程的影响。结果模型组的肌酐、肾脏纤维化的比例、24 h尿蛋白定量、VEGF/PI3K/Akt通路的miRNA蛋白的相对表达水平比对照组高(P<0.05)。芪归药对组的肌酐、空腹血糖、糖化血红蛋白数值、肾组织纤维化面积比例、尿素氮、24 h尿蛋白定量低于DN组(P<0.05),且芪归药对组糖化血红蛋白数值、空腹血糖、肌酐、尿素氮及VEGF/PI3K/Akt通路蛋白表达量与达格列净组基本相同。结论芪归药对通过调节VEGF/PI3K/Akt通路相关靶点miRNA蛋白的表达量起到抗肾脏组织纤维化及抗炎症的作用,以达到延缓DN病程进展的目的。Objective To explore the effect of Huangqi(Astragali Radix)-Danggui(Angelicae Sinensis Radix)medicinal pairs on the renal tubulointerstitial fibrosis in diabetic nephropathy(DN)rats through the VEGF/PI3K/Akt pathway,and to verify the therapeutic mechanism and curative effect of Huangqi(Astragali Radix)-Danggui(Angelicae Sinensis Radix)medicinal pairs on DN.Methods DN rat model was established by intraperitoneal injection of streptozocin(STZ),and STZ was injected at 30 mg/kg^(-1),and divided into model group and Huangqi(Astragali Radix)-Danggui(Angelicae Sinensis Radix)medicinal pairs group(Huangqi(Astragali Radix)∶Danggui(Angelicae Sinensis Radix)=1∶1,5.40 g·kg^(-1)·d^(-1)),Dapagliflozin group(0.45 mg·kg^(-1)·d^(-1)),5 rats in each group,5 SD rats of the same age as the normal control group;and the body weights of the rats in each group were measured during the administration period,urine volume,urine sugar,urine ketone,fasting blood glucose INS,and HbA1c examination was performed before administration and after the last administration.After 8 weeks,3 rats'kidneys were isolated from each group,and paraffin sections of kidney tissues were prepared according to histopathological data for observation of renal histopathology and ultrastructural morphology.The miRNA expression levels of VEGF,PI3K and Akt in renal tissues of DN rats were detected by RT-PCR.The protein expressions of VEGF,PI3K and Akt were detected by immunohistochemistry to explore the effect of Huangqi(Astragali Radix)-Danggui(Angelicae Sinensis Radix)medicinal pairs'intervention of VEGF/PI3K/Akt pathway on the progression of renal tubule interstitial fibrosis in DN rats.Results Model group creatinine,24h urinary protein quantity,ratio of kidney fibrosis,VEGF/PI3K/Akt pathway miRNA protein had a relatively high expression level higher than the control group(P<0.05).The creatinine,glycated hemoglobin values,empty blood glucose,24 h urine protein quantity,urea nitrogen,and renal tissue fibrosis area of Huangqi(Astragali Radix)-Danggui(Angelicae

关 键 词：糖尿病肾病 慢性肾脏病 黄芪当归药对 血管内皮生长因子通路(VEGF) 磷脂酰肌醇3激酶(PI3K) 蛋白激酶B(Akt) miRNA蛋白表达量 抗纤维化 

分 类 号：R285.5[医药卫生—中药学]