摘要
背景:瞬时受体电位阳离子通道蛋白6是足细胞上一种新发现的阳离子通道蛋白,是组成裂隙隔膜重要蛋白之一,其表达变化与糖尿病肾病蛋白尿密切相关。目的:观察高糖刺激对足细胞瞬时受体电位阳离子通道蛋白6表达的影响,探讨糖尿病肾病发生发展的可能机制。方法:以永生化小鼠足细胞(MPC5)作为实验对象,将其设为:正常糖组(D-葡萄糖5.6 mmol/L);渗透压对照组(D-葡萄糖5.6 mmol/L+甘露醇25 mmol/L);高糖组(D-葡萄糖30 mmol/L);缬沙坦组(D-葡萄糖30 mmol/L+10-5mol/L缬沙坦);高糖+U73122组(D-葡萄糖30 mmol/L+10μmol/L磷脂酶抑制剂U73122)。各组细胞干预时间为48 h。采用荧光定量PCR和western-blot方法检测各组足细胞瞬时受体电位阳离子通道蛋白6、nephrin、血管紧张素ⅡmR NA和蛋白的表达。结果与结论:与正常糖组相比,高糖组足细胞瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ蛋白及mR NA水平明显升高(P<0.01),nephrin m RNA及蛋白水平明显降低(P<0.01);与高糖组相比,缬沙坦组瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ的表达显著降低(P<0.05,P<0.01);高糖+U73122组瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ表达较高糖组明显下降(P<0.05,P<0.01);渗透压对照组与正常糖组之间各因子差异无显著性意义(P>0.05)。结果说明高糖可能通过血管紧张素Ⅱ-瞬时受体电位阳离子通道蛋白6反馈信号通路损伤足细胞,同时也为血管紧张素受体拮抗剂通过此通路保护足细胞而治疗糖尿病肾病的提供了一新的理论基础。
BACKGROUND：Transient receptor potential channel C6 （TRPC6） is a new and important slit diaphragm-associated protein in podocytes involved in regulating glomerular filter function. Glomerular TRPC6 expression is closely associated with proteinuria in diabetic kidney disease. OBJECTIVE： To investigate the expression of canonical TRPC6 in mouse podocytes induced by high glucose, and to explore the possible mechanism of diabetic kidney disease. METHODS：Mouse podocyte cels were cultured and divided into normal glucose group （5.6 mmol/L D-glucose）, normal control group （5.6 mmol/L D-glucose＋25 mmol/L mannitol） and experimental groups which were in the environment of high glucose （30 mmol/L）. The experimental groups included high glucose group, valsartan treatment groups （10-5 mol/L） and U73122 control group （10μmol/L U73122）. After 48 hours, the expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II （AngII） were detected respectively by real-time quantitative PCR and western blot analysis. RESULTS AND CONCLUSION：Compared with the normal control group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were markedly elevated in the high glucose group （P 〈 0.01）, while the expressions of mRNA and proteins of nephrin were decreased （P 〈 0.01）. The mRNA and proteins of TRPC6 and angiotensin II expressions were significantly down-regulated by valsartan （P 〈 0.05,P 〈 0.01）, while the mRNA and protein expressions of nephrin were effectively up-regulated （P 〈 0.05）. Compared with the high glucose group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were ameliorated in the U73122 control group. The expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II had no statistical significance between the normal control group and normal glucose group （P 〉 0.05）. Angiotensin II-TRPC6 signaling pathway may mediate high glucose-induced podocyte injury, meanwhile it provides a new theoretical basis for the treatment of diabetic kidney disease, by which the angiotensin receptor blockers can protect podocytes in diabetic kidney disease.
出处
《中国组织工程研究》
CAS
CSCD
2014年第46期7447-7451,共5页
Chinese Journal of Tissue Engineering Research
基金
山东省自然科学基金(2012ZRB01659)
项目名称:TRPC6与糖尿病肾病足细胞损伤的关系及干预治疗研究~~
