慢性马兜铃酸肾病肾间质纤维化发病机制的初步探讨  

Experimental study on renal interstitial fibrosis of chronic aristolochic acid nephropathy in rats

作　　者：高艳丽[1] 谌贻璞[1] 董鸿瑞[1] 丛笑[2] 

机构地区：[1]中日友好医院肾病中心,北京北京军区总医院肾内科100029 [2]中日友好医院肾病中心检验科,北京100029

出　　处：《中华肾脏病杂志》2005年第1期31-35,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金资助项目(No.30170429)中医药管理局科研基金资助项目(国中医药科2001ZDZX04)

摘　　要：目的制作大鼠慢性马兜铃酸肾病模型,初步探讨该模型肾间质纤维化的发病机制。方法雄性SD大鼠随机分为2组,模型组给关木通浸膏水溶液间断灌胃,对照组仅给自来水灌胃。灌胃后第4、8和12周每组各处死6只大鼠,留取两侧肾脏,应用实时定量反转录聚合酶链反应(realtimequantitativeRT-PCR)和免疫组织化学方法研究两组大鼠肾脏Ⅰ型胶原(ColⅠ)、转化生长因子-β1(TGF-β1)、结缔组织生长因子((CTGF)、纤溶酶原激活物抑制物-1(PAl-1)和金属蛋白酶组织抑制物-1(TIMP-1)的mRNA及蛋白质表达。结果模型组大鼠用药后第4周肾实质ColⅠ、TGF-β1、CTGF、PAI-1及TIMP-1mRNA表达显著上调,分别达对照组的9.31倍、5.16倍、1.79倍、8.66倍及2.54倍(P<0.01或P<0.05);第8周除TIMP-1有所升高外其余指标mRNA表达均有所下降,但与对照组比较差异仍有统计学意义(P<0.01或P<0.05);第12周全部指标mRNA表达继续下降,除CTGF外其余指标与对照组比较差异仍有统计学意义(P<0.01或P<0.05)。模型组大鼠用药后第4周ColⅠ、TGF-β1、CTGF、PAI-1及TIMP-1在肾小管间质的阳性染色面积显著扩大,分别达对照组的2.24倍、1.43倍、1.13倍、1.17倍及1.24倍(P<0.01),第8周和第12周全部指标阳性染色面积持续扩大(P<0.01)。结论慢性马兜铃酸肾病大鼠肾间质?Objective To establish a model of chronic aristolochic acid nephropathy (CAAN) in rats and to investigate the pathogenesis of its renal interstitial fibrosis.Methods Male Sprague-Dawley rats were randomly divided into two groups. One group received extract of Aristolochia manshuriensis Kom by gavage intermittently as model group. Another group received only tap water by gavage as controls. Six rats in each group were sacrificed at the end of 4th, 8th and 12th week respectively and the kidneys of each rat were separately harvested. The mRNA and protein expression of type I collagen (Col I ), transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), plasminogen activator inhibitor-1 (PAI-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) was detected by real-time quantitative RT-PCR and immunohistochemical staining respectively. Results The mRNA expression of Col I, TGF-β1, CTGF, PAI-1 and TIMP-1 in kidney tissue of the rats in model group was significantly upregulated compared to that in controls at the end of 4th week (9.31-, 5.16-, 1.79-, 8.66- and 2.54-fold, respectively) (P < 0.01 or P < 0.05). The mRNA expression of these factors except TIMP-1 was decreased at the end of 8th week (P < 0.01 or P < 0.05). Their expression was further downregulated at the end of 12th week, but still significantly higher than that in controls (P < 0.01 or P < 0.05) except CTGF. The positive staining areas of Col I , TGF-β1, CTGF, PAI-1 and TIMP-1 protein in tubulointerstitium of the rats in model group were markedly enlarged compared to those in controls at the end of 4th week (2.24-, 1.43-, 1.13-,1.17- and 1.24-fold,respectively) (P < 0.01).These positive staining areas were continuously increased at the end of 8th and 12th weeks (P < 0.01).Conclusion Overexpression of fibrogenic factors (TGF-β1、CTGF) and extracelluar matrix degradadon inhibitors (PAI-1、 TIMP-1) may be involved in the pathogenesis of renal interstitial fibrosis in CAAN.

关 键 词：慢性马兜铃酸肾病 肾间质纤维化 发病机制 RT-PCR 免疫组织化学 

分 类 号：R692[医药卫生—泌尿科学]