ZEA与DON联合染毒对CTLL-2细胞凋亡及凋亡相关蛋白Bax、Bcl-2和Caspase3等表达的影响  

The combined effects of deoxynivalenol and zearalenone on apoptosis and the expression of Bax,Bcl-2 and apoptosis-related proteins of CTLL-2

作　　者：王冰洁[1,2] 潘顺叶 李茜[1] 岑雪 邹辉[1] 顾建红[1] 袁燕[1] 刘学忠[1] 刘宗平[1,2] 卞建春 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《黑龙江畜牧兽医》2017年第4期36-41,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：江苏省普通高校自然科学基金项目(08KJD230002);江苏高校优势学科建设工程项目(PAPD);江苏省大学生实践创新训练计划项目(201511117043Z)

摘　　要：为了探讨玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合作用对免疫系统毒性影响及其毒理机制,试验以T淋巴细胞株(CTLL-2)为材料,在利用CCK-8试验确定ZEA、DON对CTLL-2细胞联合染毒浓度的基础上,研究了ZEA与DON联合染毒对CTLL-2细胞凋亡及凋亡相关蛋白Bax、Bcl-2和Caspase-3等表达的影响,试验设对照组、ZEA染毒组(5μg/m L ZEA)、DON染毒组(0.5μg/m L DON)、ZEA+DON联合染毒组(5μg/m L ZEA+0.5μg/m L DON),染毒时间为48 h,用流式细胞术检测细胞的凋亡率,Western-blot检测Bax、Bcl-2和Caspase级联反应相关蛋白的表达情况。结果表明:与对照组相比,各染毒组CTLL-2细胞的凋亡率均极显著升高(P<0.01);各染毒组细胞凋亡相关蛋白Bax/Bcl-2比值和Cleaved Cspase-3、Cleaved Caspase-8、Cleaved Caspase-9等蛋白的表达量均较对照组显著升高(P<0.05或P<0.01);联合染毒组呈现协同效应。说明ZEA与DON联合染毒在诱导免疫细胞凋亡方面发挥协同效应,它们共同存在可以发挥更强的免疫抑制效应;ZEA与DON联合染毒诱导CTLL-2细胞发生凋亡的机制与Bax/Bcl-2上调的线粒体通路密切相关。To explore the combined toxicity effects of Zearalenone （ Zearalenone, ZEA） and Deoxynivalenol （ Deoxynivalenol, DON） on the immune system and its toxic mechanism, the current study performed the combined effects of DON and ZEA on apoptosis and the expression of Bax, Bcl - 2 and apoptosis - related proteins of CTLL - 2 by using CTLL - 2 cells as material based on the CCK - 8 experiment which determined the combined exposure concentrations of ZEA and DON to CTLL - 2 cells. The study included the control group , ZEA exposure group （5 μg/mL ZEA）, DON exposure group（0. 5 μg/mL DON） and ZEA ＋ DON combined exposure group （5 μg/mL ZEA ＋ 0.5 μg/mL DON）. After exposing for 48 h, the flow cytometry was used to detect the apoptosis rate, and the Western - blot was used to detect the expression of Bax, Bcl - 2 and Caspase related proteins. Results showed that, compared with the control group, the expression ratio of the Bax/Bel - 2 decreased （P 〈 0.01 ） while that of the Cleaved Caspase- 3, Cleaved Caspase- 8 and Cleaved Caspase- 9 protein increased （P 〈0.05 or P 〈 0.01 ） for all exposure group. The combined exposure group showed the synergistic effect. Moreover, results showed that the combined exposure of DON and ZEA in inducing the apoptosis of immune cells exhibited a tend of synergistic effect, which shows a stronger immune inhibitory effect. The apoptosis mechanism of the combined exposure of DON and ZEA was closely related to the pathway of mitochondria mediated by Bax/Bcl - 2.

关 键 词：玉米赤霉烯酮(ZEA) 脱氧雪腐镰刀菌烯醇(DON) 联合染毒 CTLL-2细胞 CCK-8 细胞 凋亡 

分 类 号：Q813[生物学—生物工程] Q816