香豆素苷通过PI3K/AKT/mTOR通路保护糖尿病肾病大鼠足细胞损伤的机制研究  

Coumarin Glycosides Protects Podocyte Injury in Diabetic Nephropathy Rats through PI3K/AKT/mTOR Pathway

作　　者：鲁庆红 李益明[1] 王志芳[1] 张玲[1] 祝亮[1] LU Qinghong;LI Yiming;WANG Zhifang;ZHANG Lin;ZHU Liang(Department of Nephrology,Huanggang Central Hospital,Huanggang,Hubei,438000,China)

机构地区：[1]黄冈市中心医院肾病学科,湖北省黄冈市438000

出　　处：《医学分子生物学杂志》2022年第2期151-156,共6页Journal of Medical Molecular Biology

基　　金：湖北省卫生健康委员会2019~2020年度面上项目和血防专项(No.WJ2019M089)。

摘　　要：目的探究香豆素苷(coumarin glycosides)通过调控磷脂酰肌醇-3激酶(phosphoinositide 3-kinase,PI3K)-蛋白激酶B(AKT)-哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路对糖尿病肾病大鼠足细胞损伤的影响机制。方法采用链脲佐菌素(streptozotocin,STZ)腹腔注射构建大鼠糖尿病肾病模型,并用不同浓度的香豆素苷类化合物圆锥绣球(hydrangea paniculata,HP)提取物处理模型肾病大鼠,实验室检查大鼠血糖、尿蛋白以及内生肌酐清除率。免疫组织化学法(immunohistochemistry,IHC)检测大鼠足细胞凋亡情况,实时荧光定量逆转录聚合酶链反应(quantitative reverse transcriptase-polymerase chain reaction,qRT-PCR)检测足细胞裂孔膜蛋白Nephrin和Prodocin以及足细胞损伤标志蛋白Desmin蛋白的mRNA表达,Western印迹检测PI3K/AKT/mTOR信号通路及其磷酸化的蛋白表达。结果HP可显著改善模型大鼠的肾功能,与正常组相比,模型组的纤维连接蛋白和Ⅳ型胶原的表达明显上升,Nephrin和Prodocin的表达显著下调,Desmin的表达显著上调,PI3K/AKT/mTOR的磷酸化水平明显下降(P<0.05)。与模型组相比,随着HP浓度的增加,纤维连接蛋白和Ⅳ型胶原的表达随之下降,HP剂量依赖性地上调Nephrin和Prodocin,并下调Desmin,同时显著激活PI3K/AKT/mTOR信号通路(P<0.05)。结论香豆素苷可通过活化PI3K/AKT/mTOR信号通路,发挥减轻糖尿病肾病模型大鼠足细胞损伤的作用。Objective To explore the protective effect of coumarin glycosides on podocyte injury by regulating phosphoinositide 3-kinase(PI3K)-protein kinase B(AKT)-mammalian target of rapamycin(mTOR)signaling pathway in rats with diabetic nephropathy.Methods The rat model of diabetic nephropathy was constructed by intraperitoneal injection of streptozotocin(STZ),and different doses of coumarin glycosides extracted from hydrangea paniculata(HP),were used to treat the rat model of diabetic nephropathy.Blood glucose,urine protein and endogenous creatinine clearance rate in rats were measured.Immunohistochemistry(IHC)was applied to detect the apoptosis of podocytes in rats.Real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of podocyte split membrane proteins Nephrin,Prodocin,and Desmin,a podocyte injury marker.Western blotting was adopted to detect the expression levels of proteins in PI3K/AKT/mTOR signaling pathway and their phosphorylation.Results HP could significantly improve the renal function in the rat model.In the model group relative to the control group,the expression levels of fibronectin and typeⅣcollagen were significantly increased,and the expression levels of Nephrin and Prodocin were significantly downregulated while the expression of Desmin was significantly up-regulated,and the phosphorylation level of PI3K/AKT/mTOR was decreased significantly(P<0.05).With the dose of HP treatment increased,the expression levels of fibronectin and typeⅣcollagen were decreased,and the expression levels of Nephrin and Prodocin were up-regulated while the expression of Desmin was downregulated in a HP dose-dependent manner,and the PI3K/AKT/mTOR signaling pathways were significantly activated(P<0.05)when compared to the model group.Conclusion Coumarin glycosides can reduce the podocyte injury in rats with diabetic nephropathy by activating the PI3K/AKT/mTOR signaling pathway.

关 键 词：香豆素苷 PI3K/ AKT/ MTOR 通路 糖尿病肾病大鼠 足细胞损伤 

分 类 号：R965[医药卫生—药理学]