马兜铃酸对人肾小管上皮细胞细胞周期影响的实验研究

Effect of Aristolochic Acid on Cell Cycle of Human Kidney Cell in Vitro

目的:研究马兜铃酸(aristolochic acid,AA)对人肾小管细胞(human kidney cell,HKC)的细胞周期的影响。方法:以体外培养的HKC为研究对象,实验分为正常对照组和AA组。检测指标包括MTT 490 nm处OD值、细胞免疫组化测定的增殖核细胞抗原(proliferation cell neuclus antigen,PCNA)表达和流式细胞仪测定的细胞周期。结果:AA呈剂量正相关和时间正相关地抑制细胞增殖,特别是40mg/LAA组(0.49±0.076vs0.831±0.134,P<0.05)。10mg/LAA组PCNA表达增加[(75.7±2.1)%vs(42.2±5)%,P<0.01]。AA将HKC阻滞在S期,随着剂量的增加和时间的延长,愈加明显。结论:AA将人肾小管上皮细胞抑制在细胞周期的S期。

Objective：To study the effect of Aristolochic Acid （AA）on cell cycle of Human Kidney Cell in Vitro.Methods： HKC were cultured with or without AA. Cell viability was evaluated by using the 3-（4,5- dimethyl –thiazol -2-yl）-2,5- diphenyl-tetrazolium bromide （MTT） assay. The expression of PCNA（Proliferation cell neuclus antigen） was examined by immunol histochemistry. Flow cytometry tested the change of cell cycle.Results：The OD value in 490nm of 40 mg/L AA group was decreased（0.49±0.07 vs 0.83±0.13,P0.05）, as compared with the normal group. The number of PCNA positive cells in 10mg/L AA group is more than that of normal group[（75.74±2.11）% vs （42.23±5.03） %,P0.01]. Cell-cycle analysis revealed that HKC Were arrested in the S-phase in a dose- and time-dependent manner.Conclusion：HKC were arrested in the S-phase by AA.