摘要
目的采取离体细胞培养的方法观察氯化镉在不同时间、不同浓度作用下抑制正常大鼠肾纤维(NRK)细胞生长,及对Bcl-2、Bax基因的表达的影响。方法实验中选择5、10、20、40、60μmol/L氯化镉对NRK细胞染毒24 h,及20μmol/L氯化镉对NRK细胞染毒0.5、2、6、12、24 h,流式细胞仪观察细胞凋亡率,选择5、10、20、40、60μmol/L不同浓度的氯化镉离体培养NRK细胞12 h,及20μmol/L氯化镉分别离体培养NRK细胞0.5、2.0、6.0、12.0、24 h,利用免疫组化和逆转录聚合酶链反应(RT-PCR)方法分别检测Bcl-2、Bax蛋白及其基因的表达。结果氯化镉可以诱导NRK细胞凋亡,并且有剂量、时间-效应趋势。随着染毒剂量的增加和时间延长,Bax蛋白表达逐渐增强,且有剂量、时间-效应趋势,而Bax基因表达量保持在一个相对稳定的水平,无剂量、时间-效应趋势,Bcl-2蛋白及其基因的表达随着染镉时间的延长和浓度的增加,表达逐渐减弱,有剂量、时间-效应趋势。结论Bax蛋白表达增强,Bcl-2蛋白表达减弱是镉诱导NRK细胞凋亡过程中一个重要的调节机制,且与Bax/Bcl-2比值有密切的关系。
Objective To study the influence of cadmium chloride （ CdCl2 ） on the expression of Bcl-2, Bax and apoptosis in NRK cells in vitro at different time and concentrations. Methods NRK cells were exposed to 5,10, 20,40, 60 μmol/L CdCl2 for 24 hours and NRK cells were exposed to 20 μmol/L CdCl2 at different time points. FCM was used to measure the rate of cell apoptosis. The effects of Bcl-2, Bax protein and gene expression were measured by immunohlstochemistry technique and RT-PCR respectively. Results NRK cells apoptosis can be induced by cadmium, with a dose and time-dependent relationship. With the cadmium concentration and time increasing, Bax protein expression gradually increased and with a dose and time-dependent relationship. However, there were no statistical significance between the groups of the Bax gene expression. Bcl-2 protein and gene expression levels tend to decline with the increase of concentration and time. Conclusion One of the mechanisms of cadmium-induced apoptosis is that cadmium enhances Bax expression, and inhibites Bcl-2 expression.
出处
《中国职业医学》
CAS
北大核心
2009年第1期18-21,共4页
China Occupational Medicine
