益气养阴通络方通过抑制PI3K/AKT通路和水通道蛋白2(AQP2)表达减轻糖尿病肾病大鼠肾损伤  

Yiqi Yangyin Tongluo Recipe(YYTR)alleviates renal injury in diabetic nephropathy rats by inhibiting PI3K/AKT pathway and aquaporin 2 (AQP2)

作　　者：宇汝翠 陆智慧 李金虎 YU Rucui;LU Zhihui;LI Jinhu(Department of Traditional Chinese Medicine,The First Affiliated Hospital of University of Science and Technology of China(Anhui Provincial Hospital),Hefei 230001,China)

机构地区：[1]中国科学技术大学附属第一医院(安徽省立医院)中医科,安徽合肥230001

出　　处：《细胞与分子免疫学杂志》2022年第2期138-145,共8页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81904155)。

摘　　要：目的探讨益气养阴通络方(YYTR)对糖尿病肾组织磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)、水通道蛋白2(AQP2)表达的影响及机制。方法将60只Wistar大鼠随机分为空白组、模型组、缬沙坦(20 mg/kg)组、YYTR(4 mg/kg、1 mg/kg)组、4 mg/kg YYTR联合0.5μmol/kg 740Y-P组,每组各10只。除空白组外,其余各组大鼠利用高糖高脂饲料联合注射链脲佐菌素(STZ)制备糖尿病肾病(DN)大鼠模型。造模成功次日,灌胃给药,共8周。实验结束后检测肾功能指标,根据肾脏组织HE染色和Masson染色结果评价肾小球硬化指数、肾间质损伤指数。Western blot法检测AQP2、PI3K、AKT、磷酸化PI3K(p-PI3K)和磷酸化AKT(p-AKT)蛋白水平,免疫组织化学染色检测肾组织AQP2表达和分布。结果与正常组比较,模型组大鼠24 h尿蛋白总量(24 h UTP)、血肌酐、尿素氮、β2微球蛋白(β2-MG)含量、肾脏指数、肾小球硬化指数、肾间质损伤指数、AQP2蛋白和PI3K、AKT、p-PI3K和pAKT蛋白表达量升高;与模型组相比,4 mg/kg YYTR组大鼠以上指标均明显降低,与4 mg/kg YYTR组相比,YYTR联合740Y-P组大鼠以上指标则升高。结论YYTR下调糖尿病肾病大鼠肾脏组织AQP2蛋白表达并抑制PI3K/AKT通路活化发挥肾保护作用。Objective To explore the effects of Yiqi Yangyin Tongluo Recipe(YYTR) on the expressions of phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT) and aquaporin 2(AQP2) in diabetic renal tissue and its mechanism. Methods Sixty Wistar rats were randomly divided into the blank group, model group, valsartan(20 mg/kg) group, YYTR(4 mg/kg or 1 mg/kg), 4 mg/kg YYTR combined with 0.5 μmol/kg 740Y-P group, with 10 rats in each group. Except the blank group, the rats in other groups were fed with high-glucose and high-fat diet and injected with streptozocin to create diabetic nephropathy(DN) rat models. The day after successful modeling, the rats were administrated intragastrically for 8 weeks. At the end of the experiment, renal function indexes were measured, and glomerular sclerosis index and renal interstitial injury index were evaluated according to the results of HE staining and Masson staining. Western blot analysis was used to detect the levels of AQP2, PI3K, AKT, p-PI3K and p-AKT. Immunohistochemical staining was used to detect the expression and distribution of AQP2 in renal tissue. Results Compared with the normal group, urine protein quantitation in 24 hours(24 h UTP), serum creatinine, urea nitrogen and β2-microglobulin(β2-MG) content in model group were increased. It also reported a rise of kidney index, glomerulo sclerosis index, renal interstitial injury index, AQP2 protein and PI3K, AKT, p-PI3K and p-AKT protein expressions in model group. But compared with model group, the above indexes all decreased in 4 mg/kg YYTR group. In addition, compared with 4 mg/kg YYTR group, the above indexes in YYTR combined with 740Y-P group were increased. Conclusion YYTR can protect renal function by down-regulating AQP2 protein expression and inhibiting PI3K/AKT signaling pathway activation.

关 键 词：益气养阴通络方 糖尿病肾病 磷脂酰肌醇3激酶(PI3K) 蛋白激酶B(AKT) 肾损伤 水通道蛋白2(AQP2) 

分 类 号：R285.5[医药卫生—中药学] R392-33[医药卫生—中医学]