VEGF及TRPC6的表达与糖尿病肾病大鼠足细胞损伤的相关机制  

Correlation between expressions of VEGF and TRPC6 and their roles in podocyte injury in rats with diabetic nephropathy

作　　者：朱晓娜 王玉环[2] 吴娟娟[2] 董鹏[2] 张敏[3] ZHU Xiaona;WANG Yuhuan;WU Juanjuan;DONG Peng;ZHANG Min(Department of Nuclear Medicine;Department of Endocrinology, The Second Affiliated Hospital of Xi＇an Jiao Tong University, Xi＇an 710004, China;Department of Endocrinology, The First Affiliated Hospital of Xi＇an Medical University, Xi＇an 710077, China)

机构地区：[1]西安交通大学第二附属医院核医学科,陕西西安710004 [2]西安交通大学第二附属医院内分泌科,陕西西安710004 [3]西安医学院第一附属医院内分泌科,陕西西安710077

出　　处：《南方医科大学学报》2018年第3期296-304,共9页Journal of Southern Medical University

基　　金：西安市科技计划医学攻关项目(NO.SF1316)

摘　　要：目的探讨血管内皮生长因子(VEGF)与瞬时受体电位阳离子通道蛋白6(TRPC6)有无相关性及其相互作用的可能机制。方法随机取正常SD大鼠8只作为正常对照组,腹腔注射链脲佐菌素(STZ)65 mg/kg建立糖尿病肾病大鼠模型40只,并随机分为5组,每组8只。正常对照组及糖尿病肾病组不干预,其余4组分别给予SU5416 10 mg/kg、5 mg/kg(2次/周)以及LY294002 2 mg/kg、1 mg/kg(1次/d)腹腔注射干预8周。检测大鼠血肌酐、尿素氮及24 h尿蛋白生化指标变化情况,观察糖尿病肾病大鼠肾脏组织的病理改变,同时采用Realtime PCR及Western blot观察VEGF、Nephrin、TRPC6 m RNA表达及蛋白质的变化。结果与正常对照组相比,糖尿病肾病组大鼠空腹血糖、血肌酐、尿素氮、24 h尿蛋白升高,Nephrin的表达下降,VEGF及TRPC6的表达升高(P<0.05)。SU5416干预后24 h尿蛋白及尿素氮下降,Nephrin表达升高,VEGF表达无明显变化,TRPC6表达下降(P<0.05)。LY294002干预后24 h尿蛋白下降,Nephrin及VEGF表达无明显变化,TRPC6表达下降(P<0.05)。结论VEGF对TRPC6的调控作用可能是通过PI3K/Akt通路实现的。抑制VEGFR-2和/或者阻断PI3K/Akt信号通路可阻断VEGF对TRPC6的这种调控作用。Objective To analyze the correlation between the expressions of vascular endothelial growth factor（VEGF） and transient receptor potential canonical 6（TRPC6） and their role in podocyte injury in rats with diabetic nephropathy. Methods Forty SD rats with diabetic nephropathy induced by intraperitoneal injection of 65 mg/kg streptozotocin were randomized equally into 5 groups, including a diabetic nephropathy model group and 4 treatment groups, with 8 normal SD rats as the normal control group. In the 4 treatment groups, the rats received intraperitoneal injections with SU5416 at 5 mg/kg or 10 mg/kg twice a week or with LY294002 at 1 mg/kg or 2 mg/kg once daily for 8 weeks. Blood glucose, serum creatinine, blood urea nitrogen, and 24-h urinary protein levels of the rats were detected at different time points, and the pathologies in the renal tissue were observed using HE staining, PAS staining and immunohistochemistry. The expressions of VEGF, nephrin, and TRPC6 at m RNA and protein levels were detected using RT-PCR and Western blotting. Results Compared with normal control rats, the diabetic rats showed significantly increased fasting blood glucose, serum creatinine, blood urea nitrogen and 24-h urinary protein levels with decreased expressions of nephrin m RNA and protein（P〈0.05） and increased expressions of VEGF and TRPC6（P〈0.05）. Compared with the untreated diabetic rats, the rats with SU5416 treatment showed increased 24-h urinary protein, urea nitrogen, and nephrin expression and decreased TRPC6 expression without significant changes in fasting blood glucose, serum creatinine, or VEGF expression. The rats treated with LY294002 showed decreased 24-h urinary protein and TRPC6 expression without significant changes in fasting blood glucose, serum creatinine, urea nitrogen, or expressions of nephrin and VEGF. Conclusion The regulatory effect of VEGF on TRPC6 can be blocked by inhibiting VEGFR-2 or blocking PI3 K/Akt signaling pathway.

关 键 词：糖尿病肾病 足细胞 血管内皮生长因子 瞬时受体电位阳离子通道蛋白6 磷脂酰肌醇3激酶 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]