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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 investigate the effect of cadmium chloride (CdCl2) on the growth of normal rat kidney (NRK) cells and the expression of Bcl-2 and Bax genes under different time and different concentrations in vitro. Methods NRK cells were exposed to 5, 10, 20, 40 and 60μmol / L cadmium chloride for 24 h and 20 μmol / L cadmium chloride for 0.5, 2, 6, 12 and 24 h, respectively Cell apoptosis rates were observed by flow cytometry. NRK cells were cultured in different concentrations of cadmium chloride (5, 10, 20, 40 and 60μmol / L) for 12 h, and cultured in vitro. NRK cells were cultured in vitro with 20μmol / The expression of Bcl-2 and Bax protein and their genes were detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) method at 2.0, 6.0, 12.0 and 24 h respectively. Results Cadmium chloride could induce apoptosis of NRK cells with dose-and time-effect trend. With the increase of dose and prolongation of time, the expression of Bax protein gradually increased with dose and time-effect trend, while the expression of Bax gene remained at a relatively stable level with no dose, time-effect trend, Bcl-2 The protein and its gene expression with the cadmium exposure time and concentration increased, the expression gradually weakened, there is dose, time-effect trend. Conclusions The expression of Bax protein is increased and the expression of Bcl-2 protein is decreased. It is an important regulatory mechanism of cadmium-induced NRK cell apoptosis and closely related to the Bax / Bcl-2 ratio.