柔嫩艾美耳球虫感染对鸡血清凋亡蛋白的影响

Effect of Apoptosis Protein after Eimeria tenella Infection in Chicken

为了研究柔嫩艾美耳球虫(Eimeria tenella)感染对鸡血清细胞凋亡相关蛋白的影响,探讨柔嫩艾美耳球虫与宿主相互作用,将240只体重接近的7日龄海兰灰公雏鸡分为4组:A组为不感染不诱导凋亡组;B组为不感染诱导凋亡组;C组为感染不诱导凋亡组;D组为感染诱导凋亡组。诱导凋亡的处理方法是采血前2.5 h按6.4 mL/kg体重灌服40%酒精诱导凋亡,不诱导凋亡的处理则同时灌服与40%酒精等体积的生理盐水替代。于柔嫩艾美耳球虫感染(每只鸡感染1×10^5个柔嫩艾美耳球虫孢子化卵囊)后第24、48、72、96、120 h,每组抽取10只鸡心脏采血,制备血清,采用ELISA法检测血清中细胞色素C(Cyto-C)、半胱氨酸蛋白酶(Cas)-9、Cas-8、Cas-3、B细胞淋巴瘤/白血病(Bcl)-2、Bcl-2相关X蛋白(Bax)、Bcl-XL、BH3结构域凋亡诱导蛋白(Bid)的浓度。结果显示,感染后24、48、72、96 h D组Cyto-C、Cas-9、Cas-8、Cas-3的浓度均低于B组,感染120 h D组Cyto-C、Cas-9、Cas-8、Cas-3的浓度均显著高于B组(P<0.05),感染24、48、72、96 h D组Bcl-2/Bax和Bcl-XL/Bid值均高于B组,感染120 h D组的Bcl-2/Bax低于B组,Bcl-XL/Bid值高于B组。说明柔嫩艾美耳球虫感染早期通过Bcl-2/Bax值、Cyto-C、Cas-9、Cas-8、Cas-3浓度的增加来抑制宿主细胞凋亡,感染中晚期通过Bcl-2/Bax值、Cyto-C、Cas-9、Cas-8、Cas-3浓度的降低促进宿主细胞凋亡。

In order to study the effect of Eimeria tenella infection on apoptosis related proteins in chicken serum,and explore the interaction between Eimeria tenella and its host.2407-day-old Heilan Grey male chickens were divided into 4 groups,group A was non infection and non induction of apoptosis,group B was non infection and induction of apoptosis,group C was infection and non induction of apoptosis,group D was infection and induction of apoptosis.The treatment method of inducing apoptosis was to take 40%alcohol to induce apoptosis at 6.4 mL/kg body weight at 2.5 h before blood collection,while the treatment of not inducing apoptosis was replaced by 40%alcohol and other volumes of normal saline.At 24,48,72,96 and 120 h after Eimeria tenella infection(1×10^5 Eimeria tenella sporulated oocysts per chicken),10 chickens in each group were collected for blood collection.The concentrations of Cytochrome C(Cyto-C),Caspase(Cas)-9,Cas-8,Cas-3,Bcl-2,Bax,Bcl-XL,Bid were detected by ELISA.The results showed that the concentrations of Cyto-C,Cas-9,Cas-8 and Cas-3 in group D were lower than those in group B at 24,48,72 and 96 h after infection.The concentrations of Cyto-C,Cas-9,Cas-8 and Cas-3 in group D at 120 h were significant higher than those in group B(P<0.05).The value of Bcl-2/Bax and Bcl-XL/Bid in group D at 24,48,72 and 96 h after infection were higher than those in group B.The values of Bcl-2/Bax in group D at 120 h were lower than that in group B and the value of Bcl-XL/Bid in group D was higher than that in group B.The results showed that the apoptosis of host cells was inhibited by the increase of Bcl-2/Bax value,Cyto-C,Cas-9,Cas-8 and Cas-3 at the early stage of Eimeria tenella infection,and was promoted by the decrease of Bcl-2/Bax value,Cyto-C,Cas-9,Cas-8 and Cas-3 at the middle and late stage of infection.