摘要
目的筛选转染人转化生长因子β1(TGFβ1)基因的大鼠系膜细胞相关反应性基因,并观察TGFβ1对其中之一的酸性核糖体蛋白P0的影响。方法筛选经SSHPCR和反向杂交法构建的大鼠系膜细胞TGFβ1相关反应性基因cDNA消减杂交库,并进行测序及与GenBank资料作同源性比较;分别用NorthernBlot、半定量RTPCR观察TGFβ1、酸性核糖体蛋白P0基因在培养的大鼠系膜细胞和动物模型体内表达的变化。结果经筛选获得的28个反应性基因cDNA片段,长度为59～535bp,其中7个片段均和已知酸性核糖体蛋白P0基因同源性一致。在转染TGFβ1的大鼠系膜细胞中P0基因mRNA表达增强;在大鼠抗Thy1系膜增生性肾小球肾炎模型的病变肾组织中,P0基因mRNA和TGFβ1基因mRNA的上调表达趋势一致。结论酸性核糖体蛋白P0是一个与TGFβ1基因表达密切相关的基因,可能参与TGFβ1介导的肾小球肾炎和肾小球硬化的发生机制。
Purpose To screen the responsive genes related to transforming growth factor-β1 （TGF-β1） in rat mesangial cell （MsC） transfected by human TGF-β1 gene; To detect the expression of acidic ribosomal protein P0, one of the responsive genes,in cultured rat MsC and renal cortex of rats with anti-Thyl glomerulonephritis. Methods Human TGF-β1 gene was transferred into rat MsC by lipofectin and confirmed by Western blot;SSH-PCR and differential screening for detecting the responsive genes related to TGF-β1 in MsC; Northern blot and semi-quantification RT-PCR analysis for identifying the expression of acidic ribosomal protein P0 in MsC in vitro and vivo. Results Compared with normal MsC,it was proved 28 responsive cDNA fragments related to TGF-β1 in transfected cells. Among them,7 expression sequence tags （EST） were completely identical to data of acidic ribosomal protein P0 from GenBank. Elevated levels of acidic ribosomal protein P0 mRNA were found in transfected cells and renal cortex of rat with anti-Thyl. 1 glomerulonephritis. Conclusions Acidic ribosomal protein P0 was closely related to TGF-β1 expression. It is possible that upregulation of acidic ribosomal protein P0 may be involved in the development of glomerulonephritis and glomerulosclerosis mediated by TGF-β1.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2005年第5期509-512,521,共5页
Fudan University Journal of Medical Sciences
基金
国家自然科学基金项目(30170430)资助
