1,25(OH)_2D_3抑制嘌呤霉素氨基核苷诱导的肾病大鼠TRPC6的表达  

1,25-dihydroxyvitamin D_3 inhibits TRPC6 expression in kidney of nephrosis rat induced by puromycin aminonucleoside

作　　者：肖厚勤[1,2] 费沛[1] 陈新河[1] 胡兆雄[1] 张永[1] 刘双信[2] 史伟[2] 

机构地区：[1]湖北医药学院附属太和医院肾内科,湖北十堰442000 [2]广东省人民医院肾内科,广东广州510080

出　　处：《基础医学与临床》2014年第5期667-673,共7页Basic and Clinical Medicine

基　　金：湖北省自然科学基金重大项目(2010CDA037);湖北省教育厅重点项目(D20102105);十堰市太和医院博士启动项目(2010QD02)

摘　　要：目的观察1,25-二羟维生素D3[1,25(OH)2D3]对嘌呤霉素氨基核苷(PAN)肾病大鼠肾脏TRPC6表达的影响。方法大鼠随机分为对照组(Con)、模型组(Mod)和1,25(OH)2D3治疗组(Vit D),每组n=10。Mod组和Vit D组每10天尾静脉注射PAN 100 mg/kg建立PAN肾病模型。Vit D组每天给予1,25(OH)2D30.2μg/kg灌胃。在1和3月时分两批处死动物,检测24 h尿蛋白、肾功能、血脂和血浆蛋白;PAS染色和透射电镜观察肾组织学改变;RT-PCR、激光共聚焦检测TRPC6、synaptopodin及nephrin的表达和分布。结果 PAN肾病大鼠逐渐出现大量蛋白尿、大量腹水、高脂血症和低蛋白血症肾病综合征的表现。病理呈现肾间质水肿、炎性细胞浸润、大量的蛋白管型及局灶节段性肾小球硬化、肾小管萎缩和纤维化。与Mod组大鼠比较Vit D组大鼠24 h尿蛋白显著减少[1月时,(338±120)mg vs(669±142)mg,3月时(432±83)mg vs(601±95)mg,P<0.01],肾小球硬化指数显著减轻[(2.3+0.6)vs(3.4+0.4),P<0.01],肾功能改善[Cr(40.2±3.4)μmol/L vs(53.4±6.3)μmol/L,BNU(9.4±3.0)mmol/L vs(17.3±2.9)mmol/L,P<0.01];激光共聚焦显示PAN大鼠TRPC6表达显著升高并与synaptopodin高度融合,Nephrin mRNA的表达显著降低;与Mod组大鼠比Vit D组大鼠TRPC6 mRNA显著降低,nephrin的表达显著升高[在1月时,(0.42±0.10)vs(0.75±0.14);在3月时(0.35±0.07)vs(0.68±0.10),P<0.01],Nephrin mRNA[在1月时(0.81±0.19)vs(0.33±0.09);在3月时(0.77±0.10)vs(0.44±0.10),P<0.01]。结论 1,25(OH)2D3通过抑制PAN肾病大鼠TRPC6的表达来发挥肾脏保护作用。Objective To evaluate Effect 1,25-dihydroxyvitamin D3 [1,25 （OH）2D3] on TRPC6 expression in kidney of rat model in puromycin aminonucleoside nephropathy （PAN）.Methods Thirty male Sprague-Dawley rats were randomly divided into three groups：PAN model group （Mon）,1,25 （OH）2D3 treated group（Vit D） and normal control group （Con）.Con and Vit D rat medols were constructed by consecutive three time intravenous injection PAN of 100 mg.kg-1 body weight every ten days and Vit D rats were gavaged 1,25（OH）2D3（0.2 μ g/kg ·d）.The rats were sacrificed at one and three months respectively after PAN injection.24-hour urinary protein excretion was determined.Renal Function and blood lipid were determined by an autoanalyzer.The renal tissue morphology was observed with PAS staining by light and electron microscope.The expression of nephrin,TRPC6 mRNA were evaluated by RT-PCR.The protein expression and location of nephrin and TRPC6 were decided by confocal microscope.Results PAN administration caused heavy proteinuria,hydroperitoneum,hyperlipoidemia,hypoproteinemia and as well as loss of renal function.Inflammatory cell infiltration,renal interstitial edema,partly renal tubule atrophy,fibrosis and focal segmental gloumerular sclerosis were recorded.24 h urinary protein [one month,（338 ± 120）mg vs（669 ± 142）mg,three months （432 ± 83）mg vs（601 ± 95） mg,P ＜ 0.01]; index of gloumeruslocisin three months[（2.3 ±0.6）vs（3.4 ±0.4）,P＜0.01] and renal function[Cr（40.2 ±3.4） μmol/L vs （53.4 ± 6.3） μmol/L,BUN （9.4 ± 3.0） mmoL/L vs （17.3 ± 2.9） mmol/L,P ＜ 0.01] were significant lower in Vit D group compared with Mod group,but was not significant difference in blood fat.TRPC6 mRNA expression was increased and nephrin mRNA expression was decreased in PAN rats model.TRPC6 mRNA expression[1 month,（0.42 ±0.10） vs （0.75 ±0.14）,three months （0.35 ±0.07） vs（0.68 ±0.10）,P ＜0.01]were significant lower,and nephrin mRNA[one month,（0.81 ±0.19�

关 键 词：足细胞 1 25-二羟维生素D3 嘌呤霉素氨基核苷 TRPC6 

分 类 号：R692[医药卫生—泌尿科学]