摘要
目的 探讨马兜铃酸(AA)对体外培养的人肾小管上皮细胞株(HK-2)及人肾间质成纤维细胞(hRIFs)的作用。方法 (1)用MTT比色法检测细胞增殖反应。(2)用乳酸脱氢酶(LDH)释放试验检测AA的细胞毒作用。(3)应用RT-PCR观察细胞Ⅰ型胶原(Col Ⅰ)、转化生长因子β(TGF-β)、纤溶酶原激活物抑制物1(PAI-1)、基质金属蛋白酶1(MMP-1)及金属蛋白酶组织抑制物1(TIMP-1)的mRNA表达。结果 (1)AA在10、20和40 μg/ml时对HK-2及hRIFs无刺激增殖及细胞毒效应(P>0.05);而在80和160μg/ml时却能明显杀伤上述细胞(P<0.01)。(2)40μg/ml浓度的AA孵育细胞16 h,可显著上调HK-2的TGF-β、PAI-1和TIMP-1 mRNA表达(P<0.05),也可上调hRIFs的Col Ⅰ、TGF-β、PAI-1和TIMP-1 mRNA的表达(P<0.05);而10和20μg/ml浓度的AA未观察到上述作用。结论 80和160μg/ml AA对HK-2有明显细胞毒作用,此作用可能与急性马兜铃酸肾病发病相关;而40μg/ml AA可上调HK-2及hRIFs的TGF-β、PAI-1和TIMP-1 mRNA表达,并能上调hRIFs的Col Ⅰ mRNA表达,此作用可能与慢性马兜铃酸肾病发病相关。
Objective To investigate the effects of aristolochic acid(AA) on cell proliferation, cytotoxicity and mRNA expression of some cytokines in cultured human renal interstitial fibroblasts (hRIFs) and human renal tubular epithelial cell line (HK-2). Methods (1) The cell proliferation of hRIFs and HK-2 was determined by MTT method. (2) The cytotoxicity of A A to hRIFs and HK-2 was determined by lactate dehydrogenase (LDH) release test. (3) The mRNA expression of type Ⅰ Collagen (Col Ⅰ), transforming growth factor-β(TGF-β), plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TTMP-1) was detected semiquantitatively with reverse transcription-polymerase chain reaction (RT-PCR). Results (1) AA in the concentrations of 10, 20 and 40μg/ml did not significantly stimulate the proliferation of hRIFs and HK-2 ( P > 0. 05) and increase LDH release rates of HK-2 and hRIFs (P > 0. 05) . (2) AA in the concentrations of 80 and 160μg/ml significantly elevated LDH release rates of HK-2 and hRIFs ( P < 0. 01); LDH release rates were increased by 1.88- and 4.86-fold in HK-2 respectively, and by 1.72- and 1.94-fold in hRIFs respectively as compared to control group. (3) When stimulated by AA of 40μg/ml for 16 hours, the mRNA expression of TGF-β, PAI-1 and TIMP-1 in HK-2 was significantly upregulated ( P <0. 05) and the mRNA expression of Col Ⅰ, TGF-β, PAI-1 and TIMP-1 in hRIFs was also significantly upregulated ( P < 0. 05) . The mRNA levels of TGF-β, PAI-1 and TIMP-1 in HK-2 were increased by 1. 65-, 1. 44- and 1. 30-fold respectively, and the mRNA levels of Col Ⅰ, TGF-β, PAI-1 and TIMP-1 in hRIFs were increased by 1. 47-,1. 41-,2. 32-and 1.41-fold respectively as compared to control group. No above-mentioned effects were found in the concentrations of 10 and 20μg/ml of AA. Conclusions The cytotoxic effects of AA in the concentrations of 80 and 160μg/ml on HK-2 may be related to pathogenesis of acute AA nephropathy. AA in the concentration of 40μg/rnl can significantly upregulate mRNA expression of TGF-β, PAI-1 and TIMP-1 of hRIFs and HK-2, and also upregulate Col Ⅰ mRNA expression of hRIFs, which may be related to pathogenesis of chronic AA nephropathy.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2002年第4期266-269,共4页
Chinese Journal of Nephrology
基金
国家自然科学基金(30170429)
