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目的研究低剂量甲基叔丁基醚(MTBE)诱导中国仓鼠肺成纤维细胞(CHL)的细胞周期改变及其对凋亡的适应性反应。方法实验设6个剂量组,每组3个平行样,分别用剂量为0、0.10、0.50、2.50、10.00 g/L的MTBE预处理CHL 6 h,并以等体积磷酸盐缓冲液为正常对照,用35.00 g/L的MTBE冲击染毒18 h,收集细胞,运用流式细胞仪碘丙锭(PI)单染术检测各剂量组细胞凋亡比例及细胞周期。结果 MTBE剂量高于30.00 g/L时,CHL的存活率明显下降,选取35.00 g/L的MTBE作为后续冲击染毒剂量。以0.50～10.00 g/L的MTBE预处理CHL 6 h后,可降低随后35.00g/L的MTBE导致的细胞凋亡率的增加,并使G2期阻滞减轻,细胞产生了适应性反应。结论 MTBE在0.50～10.00g/L可以诱导CHL对35.00 g/L剂量MTBE引起的凋亡产生适应性反应。
Objective To investigate the changes of cell cycle of Chinese hamster lung fibroblasts (CHL) induced by low dose methyl tert-butyl ether (MTBE) and its adaptive response to apoptosis. Methods Six experimental groups were established. Each group consisted of 3 parallel samples. The cells were pretreated with MTBE at 0, 0.10, 0.50, 2.50 and 10.00 g / L for 6 h, respectively, and treated with an equal volume of phosphate buffer . The cells were exposed to MTBE of 35.00 g / L for 18 h. The cells were collected and the percentage of apoptotic cells and the cell cycle were measured by flow cytometry with iodine propidium iodide (PI) staining. Results When the dose of MTBE was higher than 30.00 g / L, the survival rate of CHL decreased obviously. The MTBE of 35.00 g / L was selected as the impact dose. The pretreatment of CHL with 0.50-10.00 g / L MTBE for 6 h reduced the subsequent increase of apoptosis rate induced by MTBE of 35.00 g / L, reduced the arrest of G2 phase and produced an adaptive response. Conclusion MTBE of 0.50 ~ 10.00g / L can induce CHL to 35.00 g / L dose MTBE-induced apoptosis adaptive response.