马兜铃酸对人脐静脉内皮细胞的作用

目的探讨马兜铃酸对人脐静脉内皮细胞（HUVEC）的作用。方法用马兜铃酸钠盐（AA-Na）干预体外培养原代HUVEC。用四唑盐比色法及乳酸脱氢酶释放法分别检测细胞增殖反应及细胞毒性作用。RT-PCR及酶联免疫吸附法分别检测转化生长因子β1 （TGF-β1）、纤溶酶原激活物抑制物1（PAI-1）及血小板反应蛋白1（TSP-1）的mRNA表达及上清液含量。结果10、20、40及80 mg/L浓度的AA-Na能显著抑制HUVEC增殖及有明显细胞毒效应（P〈0.01）。与对照组相比,5 mg/L的AA-Na显著上调了HUVEC的TGF-β1、PAI-1及TSP-1 mRNA和蛋白表达（P〈0.05）。结论AA-Na可上调HUVEC的TGF-β1、PAI-1及TSP-1表达,此作用可能在慢性马兜铃酸肾病小动脉管壁病变的发病中起一定作用。

Effects of aristolochic acid on human umbilical vein endothelial cells

Objective To elucidate the effects of aristolochic acid (AA) on human umbilical vein endothelial cells (HUVEC). Methods (1) The proliferation of HUVEC was determined by MTT method. (2) The cytotoxicity of AA-Na to HUVEC was determined by lactate dehydrogenase (LDH) release test. (3) HUVEC was stimulated with 5 mg/L AA-Na for different times. The mRNA and protein expression of TGF-β1, PAI-1 and TSP-1 were measured by RT-PCR and enzyme-linked immunosorbent assay (ELISA). Results (1) When the concentration of AA-Na was higher than 10 mg/L, the proliferation of HUVEC was significantly inhibited and LDH releasing from HUVEC was significantly increased (P<0.01)； (2) When HUVEC was treated with 5 mg/L AA-Na for 8~16 h, the mRNA expression of TGF-β1, PAI-1 and TSP-1 was significantly up-regulated to 1.53, 1.24 and 1.23 times respectively (P<0.05) compared with the control group； (3) When HUVEC was stimulated with 5 mg/L AA-Na, TGF-β1 was significantly increased at 24 h (P<0.01); PAI-1 and TSP-1 were also significantly enhanced at 16 h(P<0.01). Conclusions AA-Na of 5 mg/L can up-regulate mRNA and protein expression of TGF-β1, PAI-1 and TSP-1 of HUVEC, which may be related to the pathological changes of arterioles in chronic AA nephropathy.