外源性表皮生长因子对马兜铃酸-Ⅰ所致细胞增殖抑制作用的影响  

Effects of exogenous epidermal growth factor(EGF) on growth inhibition induced by aristolochic acid I(AA-I) in renal proximal tubular epithelial cells

作　　者：周娜[1] 杨莉[1] 唐嘉薇[1] 李晓玫[1] 

机构地区：[1]北京大学第一医院肾内科,北京大学肾脏病研究所,卫生部肾脏疾病重点实验室,北京100034

出　　处：《中国中药杂志》2008年第19期2222-2226,共5页China Journal of Chinese Materia Medica

摘　　要：目的:探讨加入外源性表皮生长因子(EGF)能否改善马兜铃酸-Ⅰ(AA-Ⅰ)引起的细胞增殖抑制作用。方法:以体外培养的人近端肾小管上皮细胞系(HK-2)为研究对象,在正常培养组(对照组)、AA-Ⅰ(10 mg.L-1)刺激组(AA组)和不同时机加入EGF处理组之间比较各项指标变化;用倒置显微镜对细胞形态进行观察;台盼蓝染色后细胞计数法观察细胞增殖反应;流式细胞仪分析细胞周期变化情况。结果:与对照组相比,AA-Ⅰ对细胞的毒性作用呈浓度和时间依赖性增强,AA-Ⅰ(10 mg.L-1)刺激24,48 h后对存活细胞的细胞周期分析可见G0/G1期细胞比例分别减少31.4%和87.4%,而G2/M期细胞比例逐渐增高达对照的1.32倍和3.73倍(均P<0.05)。EGF(20μg.L-1)刺激24 h后细胞数达对照组的1.36倍(P<0.05),细胞周期中G0/G1期细胞比例减少9.7%,S和G2/M期细胞比例分别增高2.9%和6.7%,表明EGF具有促进细胞周期的作用。与AA-Ⅰ组相比,无论预先(preEGF)、同时(EGF)或刺激后(postEGF)加入EGF,AA-Ⅰ所致的细胞增殖抑制现象并没有得到改善,也未能明显改善其导致的细胞周期异常。结论:AA-Ⅰ(10 mg.L-1)能够显著抑制损伤后存活HK-2细胞的增殖,使细胞周期阻滞于G2/M期;外源性EGF(20μg.L-1)能够明显促进正常细胞增殖,但不同时机加入EGF均不能改善AA-Ⅰ所致的细胞增殖抑制作用。Objective： The purpose of this study was to investigate the effects of exogenous epidermal growth factor （EGF） on the growth inhibition of renal proximal tubular epithelial cell induced by AA-Ⅰ. Method： Cultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. The changes of the survived HK-2 ceils were observed and compared among control, AA-Ⅰ stimulation, pre-EGF, together-EGF and post-EGF groups. In the study, cellular morphologic assessments were performed with a phase-contrast inverted microscope. Cell counting after stained with 0.04% trypan blue was adopted to analyze cell proliferation. Cell cycle was assessed by flow cytometry. Result： Number of the survived cells stimulated by AA-Ⅰ for 12, 24, 48 hours decreased gradually in a dose and time dependent manner. At 24, 48 hours, the survived cells showed a significant disturbance in the cell cycle procedure, which was characterized as decreased percentage of cells in G0/G1 phase, significant increased percentage of cells in G2/M phases. Exogenous EGF （20 μg· L^-1） could significantly promote the proliferation of HK-2 cells, which shown a increased cell number, accompanied down-regulated cells in G0/G1 phase, increased ceils in S and G2/M phase. Compared with AA-Ⅰ groups, it failed to improve the inhibitory effect on cell proliferation and abnormal cell cycle procedure by AA-Ⅰ, no matter EGF was added in before, at same time or after AA-Ⅰ stimulation. Conclusion： AA-Ⅰ （10 g·L^-1） has remarkable growth inhibition effects on survived RTEC, and can induce a blockage of G2/M phase in the cell cycle procedure. Exogenous EGF （20 μg·L^-1 ） promote the proliferation in normal cultured HK-2 cell. EGF treatment could not improve the proliferation inhibitory effect induced by AA-Ⅰ, no matter adding EGF to cultures before, together with AA-Ⅰ or after AA-Ⅰ stimulation.

关 键 词：马兜铃酸-Ⅰ 肾小管上皮细胞 表皮生长因子 细胞周期 

分 类 号：R285.5[医药卫生—中药学]