Xu Zhi-Yong, Zheng Ming-Xue, Zhang Yan, Cui Xiao-Zhen, Yang Sha-Sha, Liu Rui-Li, Li Shan, Xi Rou, Gong Xin, Bai Rui
College of Animal Science and Technology, Shanxi Agricultural University, Taigu, 030801, China; College of Animal Science and Technology, Henan Institute of Science and Technology, Xinxiang, 453003, China.
College of Animal Science and Technology, Shanxi Agricultural University, Taigu, 030801, China.
Exp Parasitol. 2016 Dec;171:42-48. doi: 10.1016/j.exppara.2016.10.011. Epub 2016 Oct 17.
The purpose of the present study was to investigate the dynamic changes in the main regulatory genes of the mitochondrial permeability transition pore in E. tenella host cells. Primary chick embryo cecum epithelial cell culture techniques, spectrophotometer technology, Hoechst-Annexin V-PI apoptosis staining and ELISA were used to detect the apoptosis rate and dynamic changes of Bcl-2, Bcl-xl, Bax, Bak, Bid, Bad, HK-II, and ATP content in E. tenella host cells at 4, 24, 48, 72, 96, and 120 h. The rates of early apoptosis, late apoptosis, and necrosis of group T0 were significantly lower (P < 0.05) or highly significantly lower (P < 0.01) than those of group C at 4 h, but higher (P < 0.05 or P < 0.01) at varying degrees than those of the same group at 24-120 h. Compared to group C, the amount of Bcl-2, ATP, Bax and Bad in group T0 were visibly lower (P < 0.05 or P < 0.01) at 4 h, whereas Bcl-xl/Bax was highly significantly higher (P < 0.01) at 4 h. In addition, group T0 had less ATP at 24-120 h than group C, whereas the amount of Bcl-2, Bcl-xl, Bax, Bak, Bid, Bad and HK-II in group T0 inversely increased in varying degrees at 24-120 h compared with group C. Moreover, Bcl-2/Bax was lower (P < 0.01) at 24, 48, and 96 h, and Bcl-xl/Bax was lower (P < 0.05) at 48 h in group T0 than in group C, respectively. Taken together, these observations indicate that in the early developmental stages of E. tenella, the host-cell apoptosis rate decreased; although the amount of anti- and pro-apoptotic genes in host cells decreased, the ratios of anti-apoptotic to pro-apoptotic bcl-2 gene-family members increased. In the middle and later developmental stages of E. tenella, the host-cell apoptosis rate increased; the amount of anti- and pro-apoptotic genes increased, while the ratios of anti-apoptotic to pro-apoptotic bcl-2 gene-family members decreased. In addition, ATP decreased at all developmental stages of E. tenella.
本研究的目的是探讨柔嫩艾美耳球虫宿主细胞中线粒体通透性转换孔主要调控基因的动态变化。采用原代鸡胚盲肠上皮细胞培养技术、分光光度计技术、Hoechst-Annexin V-PI凋亡染色法及ELISA法，检测柔嫩艾美耳球虫宿主细胞在4、24、48、72、96和120 h时的凋亡率以及Bcl-2、Bcl-xl、Bax、Bak、Bid、Bad、HK-II和ATP含量的动态变化。T0组在4 h时的早期凋亡、晚期凋亡和坏死率均显著低于（P<0.05）或极显著低于（P<0.01）C组，但在24120 h时不同程度地高于（P<0.05或P<0.01）同组4 h时的水平。与C组相比，T0组在4 h时Bcl-2、ATP、Bax和Bad的含量明显降低（P<0.05或P<0.01），而Bcl-xl/Bax在4 h时极显著升高（P<0.01）。此外，T0组在24120 h时的ATP含量低于C组，而T0组在24~120 h时Bcl-2、Bcl-xl、Bax、Bak、Bid、Bad和HK-II的含量与C组相比不同程度地呈反向增加。而且，T0组在24、48和96 h时的Bcl-2/Bax低于C组（P<0.01），在48 h时的Bcl-xl/Bax低于C组（P<0.05）。综上所述，这些观察结果表明，在柔嫩艾美耳球虫的早期发育阶段，宿主细胞凋亡率降低；尽管宿主细胞中抗凋亡和促凋亡基因的含量减少，但抗凋亡与促凋亡bcl-2基因家族成员的比例增加。在柔嫩艾美耳球虫发育的中后期，宿主细胞凋亡率升高；抗凋亡和促凋亡基因的含量增加，而抗凋亡与促凋亡bcl-2基因家族成员的比例降低。此外，在柔嫩艾美耳球虫的所有发育阶段ATP含量均降低。
