【摘要】 目的 研究堆型艾美尔球虫延伸因子1-α(EaEF-1α)在堆型艾美尔球虫的感染期间调控细胞凋亡的主要机制。方法 从球虫裂殖子侵染DF-1细胞实验的上清液中分析筛选得到EaEF-1α,与质粒pEGFP-N1重组得到重组质粒pEGFP-N1-EaEF-1α,转染入DF-1细胞中，Western bolt及荧光检测蛋白表达，通过流式细胞术检测脂多糖组、pEGFP-N1组、pEGFP-N1-EF-1α组和空白组DF-1细胞凋亡情况，酶联免疫吸附实验检测脂多糖组、pEGFP-N1组、pEGFP-N1-EF-1α组和空白组DF-1细胞中凋亡因子Bax、Bcl-2、Caspase3、Caspase8表达水平。结果 流式细胞术检测发现pEGFP-N1-EF-1α组细胞凋亡率显著低于pEGFP-N1组和脂多糖组。DF-1细胞裂解液中Bax表达水平降低，Bcl-2及Caspase3表达水平上调，pEGFP-N1-EF-1α组Bcl-2/Bax的值显著高于pEGFP-N1组和脂多糖组。Caspase8表达水平无差异。结论 EaEF-1α体外过表达通过线粒体通路显著抑制细胞凋亡。更多还原

【Abstract】 Objective To investigate the main mechanism by which Eimeria acervulina elongation factor 1-α(EaEF-1α) regulates apoptosis during Eimeria acervulina infection.Method EaEF-1α was isolated from the supernatant of Eimeria acervulina schizozoites infecting DF-1 cells. It was then recombined with the plasmid pEGFP-N1 to obtain the recombinant plasmid pEGFP-N1-EaEF-1α, which was transfected into DF-1 cells. Western blot and fluorescence detection were used to verify the expression. Apoptosis of DF-1 cells in the lipopolysaccharide group, pEGFP-N1 group, pEGFP-N1-EF-1α group, and blank group was detected by flow cytometry. The expression levels of apoptosis factors Bax, Bcl-2, Caspase3, and Caspase8 in DF-1 cells in the lipopolysaccharide group, pEGFP-N1 group, pEGFP-N1-EF-1α group, and blank group were detected by enzyme-linked immunosorbent assay(ELISA). Result Flow cytometry showed that the apoptosis rate of the pEGFP-N1-EF-1α group was significantly lower than that of the pEGFP-N1 group and the lipopolysaccharide group. The expression level of Bax in the lysates of DF-1 cells decreased, while the expression levels of Bcl-2 and Caspase3 increased. The value of Bcl-2/Bax in the pEGFP-N1-EF-1α group was significantly higher than that in the pEGFP-N1 group and the lipopolysaccharide group. There was no difference in the expression level of Caspase8.Conclusion Over-expression of EaEF-1α in vitro significantly inhibits apoptosis through the mitochondrial pathway.更多还原