关键词: 堆型艾美尔球虫, 延伸因子 1-α, 线粒体通路, 细胞凋亡

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.

Key words: Eimeria acervulina, extension factor 1-α, mitochondrial pathway, apoptosis