马兜铃酸对人肾小管上皮细胞细胞周期影响的实验研究

摘要

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(prolifera tion 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40 mg/L AA组(0.49±0.076 vs 0.831±0.134,P<0.05).10 mg/L AA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 ,P < 0. 05 ), as compared with the normal group. The number of PCNA positive cells in lOmg/L AA group is more than that of normal group[ ( 75.74 ± 2. 11 )% vs ( 42. 23 ± 5. 03 ) % ,P < 0. 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.