CHIP调节UUO再通大鼠肾组织TGF-β1/Smads信号通路的研究  

CHIP Protein Regulating TGF-β Signal Pathway in the Reverse of Unilateral Ureteral Occlusion

作　　者：董飞侠[1] 张新志[2] 吴锋[3,4] 黄迪[3,4] 何立群[3,4] 

机构地区：[1]浙江中医药大学附属温州中医院肾内科,温州325000 [2]上海市第六人民医院金山分院肾内科,上海201500 [3]上海中医药大学附属曙光医院肾内科,肝肾疾病病证教育部重点实验室 [4]上海市高校中医内科E-研究院,上海201203

出　　处：《中华中医药学刊》2012年第2期245-248,I0003,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30873259);上海市优秀学科带头人培养计划资助项目(08XD14039);上海市教育委员会E-研究院建设计划资助项目(E03008);上海市教委高校创新团队资助项目;国家中医药管理局重点学科资助项目(30301041210);温州市科技局资助项目(Y20070049);温州市卫生局资助项目(2010A012);浙江省151人才和温州市551人才专项资助项目

摘　　要：目的:利用基因芯片研究Carboxyl terminus of Hsc70-interacting protein(CHIP)对肾间质纤维化TGF-β1/Smads信号通路的抑制性调节。方法:将18只SPF级SD大鼠中6只行unilateral ureteral occlusion(UUO)模型,6只植入弹性管脂肪垫加压梗阻,7天后再通定为reverse of unilateral ureteral occlusion(RUUO)组,6只UUO在7天处死留取左肾组织,另外6只为假手术组。6只RUUO大鼠在再通7天后处死,留取左侧肾组织,运用West-ern blotting检测肾组织CHIP的表达,免疫组化检测肾组织中α-SMA、CD68的阳性表达面积,定制芯片研究TGF-β1/Smads信号通路的基因变化。结果:UUO组TGF-β1、smad2、smad3、COL1a2、JunB基因显著上调,和假手术组比较上调超过1.5倍,肾组织中的α-SMA、CD68表达面积和假手术组比较有显著统计学意义P<0.01。而RUUO组CHIP表达量较UUO组更是显著增强P<0.01,TGF-β1、smad2、smad3、COL1a2、JunB基因显著下调,和UUO组比较下调超过1.5倍,肾组织中的α-SMA、CD68表达面积减小和UUO组比较有显著统计学意义P<0.01。结论:发现CHIP的过量表达能显著降低细胞内,TGF-β1、Smad2/3、COL1a2、JunB基因水平,Western blot结果也表明CHIP蛋白表达能明显降低Smads介导的基因转录活性,进一步的基因芯片结果显示CHIP蛋白能明显下调TGF-β1所诱导的下游基因JunB的表达。结果提示CHIP蛋白能抑制性调节TGF-β1/Smads信号通路,抑制肾纤维化。Objective： To investigate the role of CHIP expression inhibiting TGF -βsignal pathway by customized array in the release of unilateral ureteral occlusion（RUUO）. Methods ： 18 specified - pathogens free SD rats, six of them were underwent unilateral ureteral occlusion（UUO） operation, six of them embedding flexible tubing fat pad forcing obstruction were underwent RUUO operation after 7d, six of them were underwent sham - operation. Another six of them were for the shamoperated group. UUO rats were sacrificed after 7d, RUUO rats were harvested after 7d, specimens of the left kidney tissue, Western blotting was to detect the expression of CHIP and immunohistochemistry detection was for α - SMA, CD6s positive expression area in the release of kidney tissue, and customed array was used to detect fold up - regulation or down- regulation gene of TGF- β1/Smads signal pathway. Results： The expressions of TGF- β1 ,smad2, smad3, Coll a2 ,JunB in UUO group are increased significantly , compared with sham - operation group, which is over 1.5 times, and an positive area of α -SMA and CD6s in nephridial tissue is increased obviously in UUO group （ P 〈 0.01 ）. Compared with UUO group, the expression of CHIP increased obviously in RUUO group on 7d （ P 〈0.01 ） ,and the expressions of TGF- B1, smad2, smad3. Colla2.JunB in RUUO RUUD are decreased significanflv, comnared with UUO group,which is over 1.5 times, an positive area of α - SMA and CD68 in nephridial tissue is decreased obviously in RUUO group （ P 〈 0.01 ）. Conclusions ： It was observed that increasing expression of CHIP dramatically reduced the endougenous TGF- β1 ,Smad2/3 ,Colla2 ,JunB gene levels. Western reporter analysis showed that CHIP inhibited the Smads - induced transcription activity. Furthermore, Western blot resuhs indicate that CHIP could decrease gene expression of JunB, a quick response gene induced by TGF - β. The results suggest that CHiP can inhibit the protein regulator of TGF - β1/Smads signale pathway and inhib

关 键 词：UUO动物模型 RUUO动物模型 CHIP CD68 TGF-β1/Smads 肾间质纤维化 

分 类 号：R334.1[医药卫生—人体生理学]