基于Wnt/β-catenin/TRPC6/CaN通路探讨淫羊藿苷对激素抵抗性肾病综合征大鼠泼尼松抵抗的作用机制  

Discussion on the Mechanism of Icariin in Improving Prednisone Resistance in Rats with Steroid-Resistant Nephrotic Syndrome Based on Wnt/β-catenin/TRPC6/CaN Pathway

作　　者：白俊嫄 戴恩来[2] 蒲晓薇 张云霞[2] BAI Junyuan;DAI Enlai;PU Xiaowei;ZHANG Yunxia(Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730020,China;Gansu University of Chinese Medicine,Lanzhou 730020,China)

机构地区：[1]甘肃中医药大学附属医院,甘肃兰州730020 [2]甘肃中医药大学,甘肃兰州730020

出　　处：《中国中医药信息杂志》2024年第5期85-91,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82160852)。

摘　　要：目的观察淫羊藿苷对激素抵抗性肾病综合征(SRNS)大鼠泼尼松抵抗的改善作用,及对Wnt/β-catenin/TRPC6/CaN通路表达的影响。方法将50只SD大鼠分为空白组、模型组、泼尼松组(6.3 mg/kg)、淫羊藿苷组(50 mg/kg)和联合组,每组10只。采用2次尾静脉注射阿霉素建立SRNS大鼠模型,分别予相应方式干预6周。观察大鼠一般状态,检测24 h尿蛋白定量(24 h-UTP)、尿17-羟皮质类固醇(17-OHCS)及血清生化指标,Masson染色观察肾组织病理形态及超微结构,RT-PCR、Westernblot检测肾组织Wnt/β-catenin/TRPC6/CaN通路相关基因及蛋白表达。结果与空白组比较,模型组大鼠体质量、进食量、饮水量均明显减少,24 h-UTP增加、尿17-OHCS含量减少(P<0.01),血肌酐(SCr)、血尿素氮(BUN)、血清总胆固醇(TC)、三酰甘油(TG)含量升高(P<0.01),血清总蛋白(TP)、白蛋白(ALB)含量降低(P<0.05,P<0.01);肾小球增大,基底膜增厚,球囊壁增厚伴纤维化,足细胞空泡变性,有大量胶原纤维沉积,胶原容积分数(CVF)增加(P<0.01);Wnt、β-catenin、TRPC6、CaM、CaN、NFAT1 mRNA和蛋白表达均明显升高(P<0.01),F-actin、MYH9mRNA和蛋白表达明显降低(P<0.01)。与模型组比较,联合组大鼠体质量、饮水量明显增加,24 h-UTP降低、尿17-OHCS含量增加(P<0.01),SCr、BUN、TG含量降低(P<0.01);肾组织纤维化程度减轻,CVF减少(P<0.01),足突融合现象明显好转,足细胞结构趋于正常;肾组织Wnt、β-catenin、TRPC6、CaM、CaN、NFAT1 mRNA和蛋白表达明显降低,F-actin、MYH9 mRNA和蛋白表达明显升高(P<0.01,P<0.05)。结论淫羊藿苷可能通过抑制Wnt/β-catenin/TRPC6/CaN通路,上调骨架蛋白F-actin、MYH9基因和蛋白表达,进而保护足细胞结构、减缓足细胞损伤,改善SRNS大鼠泼尼松抵抗。Objective To observe the improving effects of icariin on the improvement of prednisone resistance in steroid resistant nephrotic syndrome(SRNS)rats and its effects on the Wnt/β-catenin/TRPC6/CaN pathway.Methods Totally 50 SD rats were divided into blank group,model group,prednisone group(6.3 mg/kg),icariin group(50 mg/kg)and combination group,with 10 rats in each group.Twice tail vein injections of adriamycin were used to establish a SRNS rat model,and corresponding interventions were given for 6 weeks.General condition of rats was observed,24-hour urinary protein quantification(24 h-UTP),urine 17-hydroxycorticosteroids(17-OHCS),and serum biochemical indicators were detected,renal tissue pathological morphology and ultrastructure were observed by Masson staining,RT-PCR and Western blot were used to detect Wnt/β-catenin/TRPC6/CaN pathway related genes and protein expressions in renal tissue.Results Compared with the blank group,the body mass,feed intake and water intake of rats in the model group significantly decreased,24 h-UTP increased,urine 17-OHCS content decreased(P<0.01),the contents of serum creatinine(SCr),blood urea nitrogen(BUN),total cholesterol(TC)and triglycerides(TG)inserum increased(P<0.01),while the contents of total protein(TP)and albumin(ALB)decreased(P<0.05,P<0.01);glomerular enlargement,thickening of basement membrane,thickening of balloon wall with fibrosis,vacuolar degeneration of podocytes,and deposition of a large amount of collagen fibers appeared,collagen volume fraction(CVF)increased(P<0.01);Wnt,β-catenin,TRPC6,CaM,CaN and NFAT1 mRNA and protein expressions significantly increased(P<0.01),while the mRNA and protein expressions of F-actin and MYH9 were significantly reduced(P<0.01).Compared with the model group,rats in the combination group showed a significant increase in body mass and water intake,a decrease in 24 h-UTP,and an increase in urine 17-OHCS content(P<0.01);the contents of SCr,BUN and TG decreased(P<0.01);the degree of renal tissue fibrosis was reduced,CVF was reduced(P<0

关 键 词：激素抵抗性肾病综合征 淫羊藿苷 足细胞 阿霉素肾病 Wnt/β-catenin/TRPC6/CaN通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]