摘要
目的:观察结缔组织生长因子(CTGF)在单侧输尿管梗阻(UUO)大鼠模型中的动态表达,探讨CTGF在肾小管间质纤维化中的作用机制。方法:将48只Wistar大鼠随机分为UUO组和假手术(SO)组,采用左输尿管结扎术复制UUO模型,于术后1、3、7、14d分别处死2组大鼠取左肾。采用Masson染色评定肾小管间质损伤程度;逆转录-聚合酶链式反应(RT-PCR)方法检测肾组织转化生长因子-β1(TGF-β1)、CTGF、Ⅰ型胶原(ColⅠ)和纤溶酶原激活物抑制物-1(PAI-1)mRNA表达;免疫组织化学方法测定TGF-β1、CTGF、ColⅠ、PAI-1表达;Westernblotting方法检测CTGF蛋白表达量的变化。结果:UUO术后1d,梗阻肾TGF-β1mRNA表达开始升高,第3-14d时升高更显著(P<0.01),CTGF、ColⅠ、PAI-1mRNA水平随之逐渐升高。免疫组织化学染色发现,UUO大鼠肾小管和间质区域CTGF表达随病程进展逐渐增强;相关分析显示,UUO术后第7d,CTGF表达量与肾小管间质损伤指数、肾小管间质TGF-β1、ColⅠ、PAI-1表达强度均呈正相关,相关系数(r)分别为0.62、0.85、0.78和0.76(均P<0.01)。Westernblotting结果显示,CTGF蛋白水平在术后第3d开始上升,随病程进展更显著。结论:CTGF可通过促进细胞外基质产生和抑制细胞外基质降解双重途径诱导肾小管间质纤维化的发生和进展。
AIM： To observe the expression of connective tissue growth factor （CTGF） in unilateral ureteral. obstruction （UUO） rats, and to explore its pathogenic role in renal tubulointerstitial fibrosis. METHODS： 48 Wistar rats were randomly divided into sham - operated and UUO group. The rats were sacrificed at day 1,3, 7, and 14. The degree of tubulointerstitial damage was scored according to the Masson staining. The mRNA and protein levels of CTGF, transforming growth factor - β1 （ TGF - β1 ）, collagen Ⅰ（ Col Ⅰ）, and plasminogen activator inhibitor - 1 （ PAl - 1 ） were detected by reverse transcriptional - pelymerase chain reaction （ RT - PCR） and immunohistoehemistry, respectively. Expression of CTGF protein in the kidney was also assessed using Western blotting. RESULTS ： TGF - β1 mRNA level began to increase as early as 1 day after UUO. This increase was followed by the elevation of CTGF mRNA level, which began to increase at third days after UUO （ P 〈 0. 01 ）. 7 days after UUO, the protein level of CTGF was positively related to the index of tubulointerstitial damage （ r = 0. 62, P 〈 0. 01 ）, the expression of TGF - β1 （ r = 0. 85, P 〈 0. 01 ）, Col Ⅰ（ r = 0. 78, P 〈0. 01 ）, and PAI - 1 （ r =0. 76, P 〈0. 01 ）, as determined by immunohistoehemistry. CTGF protein level appeared to increase progressively throughout the time course. CONCLUSION： As a downstream mediator of TGF - β1, CTGF may play a crucial role in the progression of tubulointerstitial fibrosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第9期1786-1790,共5页
Chinese Journal of Pathophysiology
基金
湖北省科技攻关计划资助项目(No.2003AA301C14)
关键词
结缔组织生长因子
转化生长因子Β
胶原Ⅰ型
纤溶酶原激活物抑制物Ⅰ
肾纤维化
Connective tissue growth factor
Transforming growth factor beta
Collagen type Ⅰ
Plasminogen activator inhibitor 1
Renal fibrosis
