甲基-β-环糊精抑制AngⅡ诱导肾小球系膜细胞增殖及TRPC6表达  

MβCD inhibits glomerular mesangial cell (GMC) proliferation and expression of TRPC6 induced by Ang Ⅱ

作　　者：张娜[1] 纪泽泉[1] 

机构地区：[1]广州医科大学第二附属医院,510260

出　　处：《实用医学杂志》2013年第12期1897-1900,共4页The Journal of Practical Medicine

基　　金：广东省自然科学基金(编号:S2012010008588);广东省教育厅高等学校科技创新项目(编号:2012KJCX0088);广州市高校科研项目(编号:10A254)

摘　　要：目的:探讨MβCD(甲基-β-环糊精)对AngⅡ诱导肾小球系膜细胞(GMC)TRPC6的表达及小窝(caveolae)在GMC增殖中的作用。方法:实验分为:空白对照组(Con组)、AngⅡ组、MβCD预处理组(MβCD组)。根据预实验选定AngⅡ10-7mol/L作用GMC24h作为AngⅡ组。MβCD组用10-2mol/LMβCD预处理GMC1h后,用10-7mol/LAngⅡ刺激24h。四甲基偶氮唑盐比色法(MTT法)检测GMC增殖,WesternBlot检测TRPC6蛋白,实时荧光定量PCR检测TRPC6mRNA表达,免疫荧光检测TRPC6的分布。结果:AngⅡ组较Con组明显促进GMC增殖(P<0.05),MβCD组明显抑制AngⅡ对GMC的增殖作用,抑制率为48.96%(P<0.01)。AngⅡ组TRPC6mRNA及蛋白水平均较Con组升高(P<0.01,P<0.05),而MβCD组TRPC6mRNA及蛋白水平均较AngⅡ组降低(P<0.01)。GMCOD值与TRPC6蛋白表达水平呈正相关(R=0.907,P<0.01)。结论:AngⅡ诱导GMC增殖需要完整的小窝结构,TRPC6与GMC增殖密切相关。Objective The aims of this study were to explore the effect of MβCD on the expression of TRPC6 in glomerular mesangial cell induced by Ang Ⅱ and the effect of caveolae on Ang Ⅱ induced GMC proliferation. Methods The ceils were divided into three groups： Control group, Ang Ⅱ group and MβCD group. The GMCs in Ang Ⅱ group were treated with 10^-7 mol/L of Ang Ⅱ for 24 h. The GMCs in MβCD group were pre- treated with 10^-2 mol/L of MβCD for 1 h, and then treated with 10^-7 mol/L of Ang Ⅱ for 24 h. The methyl thiazolil tetracolium （MTT） assay was used to detect GMC proliferation. The expression of TRPC6 protein was detected using western blot. The mRNA expression of TRPC6 was assessed using real time polymerase chain reaction （PCR）. The distribution of TRPC6 was detected using immunofluorescence. Results Ang Ⅱ significantly promoted GMC proliferation（P 〈 0.05）. MβCD markedly blocked Ang Ⅱ induced GMC proliferation at the rate of 48.96% （P 〈 0.01 ）. Compared with control group, the mRNA and protein expressions of TRPC6 significantly increased in AngⅡ group （P 〈 0.01, P 〈 0.05）. However, MβCD decreased the mRNA and protein levels of TRPC6 （P 〈 0.01 ）. The OD value of GMC had a positive correlation with protein level of TRPC6 （R = 0.907, P 〈 0.01）. Conclusion The intact caveolae structure was required for Ang Ⅱ induced GMC proliferation. Meanwhile, the expression of TRPC6 was consistent with GMC proliferation.

关 键 词：小窝 瞬时受体电位阳离子通道蛋白6 血管紧张素Ⅱ 肾小球系膜细胞 

分 类 号：R692[医药卫生—泌尿科学]