中国兽医杂志2026，Vol.62Issue(5)：11-18,8.DOI:10.20157/j.cnki.zgsyzz.2026.05.002
堆型艾美耳球虫棒状体蛋白17对鸡胚成纤维细胞凋亡的影响
Effect of Eimeria acervulina Rhoptry Organelle Protein 17 on Apoptosis in Chicken Embryo Fibroblasts
摘要
Abstract
To investigate the mechanism by which Eimeria acervulina rhoptry organelle protein 17(Ea ROP17)affects apoptosis in chicken embryo fibroblast(DF-1)cells,a eukaryotic expression vector Ea ROP17-pEGFP-N1 was constructed and transfected into DF-1 cells,forming the recombinant plasmid transfection(Ea ROP17-pEGFP-N1)group.The Blank control(CTRL)group and Empty vector transfection(pEGFP-N1)group were also established.Green fluorescence was observed under an inverted fluorescence microscope,and Western blot(WB)analysis was used to verify protein expression.DF-1 cells were then divided into four groups:CTRL group,Recombinant plasmid-treated(ROP17+LPS)group,Empty vector-treated(EGFP-N1+LPS)group,and Model(LPS)group.Flow cytometry was used to detect apoptosis rates in the four groups,and real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to examine the apoptosis pathway regulated by Ea ROP17.The results showed that the Ea ROP17-pEGFP-N1 eukaryotic expression vector was successfully constructed.Under the fluorescence microscope,green fluorescence was observed in both the Ea ROP17-pEGFP-N1 and pEGFP-N1 groups but not in the CTRL group.WB analysis confirmed successful transfection and expression of the recombinant Ea ROP17-pEGFP-N1 plasmid in DF-1 cells.The apoptosis model was successfully established,and flow cytometry results showed that,compared with the LPS group,the total apoptosis rate in the ROP17+LPS group was significantly reduced(P<0.05).RT-qPCR analysis revealed no significant difference in the expression of cytochrome c(Cyt c)mRNA between the ROP17+LPS and LPS groups(P>0.05),while the mRNA expression levels of Caspase-3,Caspase-8,and Caspase-9 were significantly downregulated(P<0.05,P<0.001,or P<0.000 1).These findings suggest that Ea ROP17 can inhibit host cell apoptosis,likely via the death receptor signaling pathway rather than the mitochondrial pathway.This study provides a scientific reference for the studies on the pathogenic mechanisms of avian coccidia.关键词
堆型艾美耳球虫/棒状体蛋白17/鸡胚成纤维细胞/细胞凋亡/死亡受体信号通路Key words
Eimeria acervulina/rhoptry organelle protein 17/DF-1/apoptosis/death receptor signaling pathway分类
农业科技引用本文复制引用
郭璐嘉,安健,李治翰,尹徽,王黎霞..堆型艾美耳球虫棒状体蛋白17对鸡胚成纤维细胞凋亡的影响[J].中国兽医杂志,2026,62(5):11-18,8.基金项目
北京农业职业学院科技创新项目(XY-YF-24-05) （XY-YF-24-05）
北京农学院教改项目(BUA20211G44) （BUA20211G44）
