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Roles of TNF receptor-associated and Fas-associated death domain proteins in the apoptosis of Eimeria tenella host cells
Veterinary Parasitology ( IF 2.6 ) Pub Date : 2021-01-07 , DOI: 10.1016/j.vetpar.2021.109351
ZhiYong Xu 1 , MingXue Zheng 2 , Yan Liu 3 , Li Zhang 2 , XueSong Zhang 2 , Rui Bai 2
Affiliation  

The present study aimed to investigate the effects of death receptor adapter proteins, namely, TNF receptor-associated death domain (TRADD) and Fas-associated death domain (FADD) proteins, on Eimeria tenella-induced host cell apoptosis. Gene silencing, culture technique for primary chick embryo cecal epithelial cells, enzyme-linked immunosorbent assay, Hoechst–Annexin V/PI apoptosis staining, fluorescence quantitative PCR, and flow cytometry were used to detect the E. tenella host cell apoptotic rate, RIP1 and FADD protein expression levels, and caspase-8 activity of the TRADD siRNA-treated and FADD siRNA-treated groups. Results showed that the apoptotic rate in the TRADD siRNA group was significantly higher than that in the NC siRNA group at 4 h post-infection with E. tenella (P < 0.05). The RIP1 protein expression level in the TRADD siRNA group was significantly lower than that in the NC siRNA group at 4–24 h (P < 0.05). The FADD expression and apoptotic rates in the TRADD siRNA group were significantly lower than those in the NC siRNA group at 24–120 h (P < 0.05). The caspase-8 activity and apoptotic rates in the FADD siRNA group were significantly lower than those in the NC siRNA group (P < 0.05) at 24–120 h. These findings indicated that E. tenella inhibited the host cell apoptosis through the TRADD-RIP1 pathway at the early developmental stage and promoted host cell apoptosis via the TRADD-FADD-caspase-8 apoptotic pathway at the middle and late developmental stages.



中文翻译:

TNF受体相关和Fas相关死亡域蛋白在艾美球虫宿主细胞凋亡中的作用

本研究旨在研究死亡受体衔接蛋白,即 TNF 受体相关死亡域 (TRADD) 和 Fas 相关死亡域 (FADD) 蛋白对艾美球虫诱导的宿主细胞凋亡的影响。采用基因沉默、原代鸡胚盲肠上皮细胞培养技术、酶联免疫吸附试验、Hoechst-Annexin V/PI 细胞凋亡染色、荧光定量 PCR 和流式细胞术检测E.tenella宿主细胞凋亡率、RIP1 和TRADD siRNA处理组和FADD siRNA 处理组的FADD 蛋白表达水平和 caspase-8 活性。结果表明TRADD中的细胞凋亡率感染E.tenella 4 h后,siRNA组显着高于NC siRNA组(P  < 0.05)。TRADD siRNA组RIP1蛋白表达水平在4-24 h显着低于NC siRNA组(P  < 0.05)。TRADD siRNA组FADD表达和凋亡率在24-120 h显着低于NC siRNA组(P  < 0.05)。 在24-120小时,FADD siRNA组的caspase-8活性和凋亡率显着低于NC siRNA组(P < 0.05)。这些发现表明E. tenella 在发育早期通过TRADD-RIP1通路抑制宿主细胞凋亡,在发育中后期通过TRADD-FADD-caspase-8凋亡通路促进宿主细胞凋亡。

更新日期:2021-01-18
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