兔脑炎原虫致脑细胞凋亡的信号转导通路的研究  

Apoptotic signal transduction pathway in cerebral tissue caused by Encephalitozoon cuniculi

作　　者：潘耀谦 王选年 岳锋 李鹏 吴玉萍 刘兴友 PAN Yao-qian;WANG Xuan-nian;YUE Feng;LI Peng;WU Yu-ping;LIU Xing-you(School of Life Science and Technology,Xinxiang University,Xinxiang,Henan 453003,China)

机构地区：[1]新乡学院生命科学技术学院

出　　处：《中国兽医学报》2019年第11期2190-2194,2214,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31372407);河南省现代农业产业技术体系资助项目(S2012-06-02);河南省科技开放合作资助项目(182106000043)

摘　　要：为探讨兔脑炎原虫引起脑组织细胞凋亡的信号转导通路,本试验用普通染色、特殊染色、免疫组化染色和Western blot检测等方法,对40只病兔和10只对照兔的脑组织进行了研究。结果显示,病兔的脑组织中均检出了形态不一的肉芽肿和脑炎原虫。兔脑炎原虫可引起巨噬细胞和肉芽肿中的上皮样细胞表达TNF-α,anti-TNF-α抗体染色呈阳性反应。细胞凋亡信号转导通路的关键蛋白caspase-8和caspase-3在血管内皮细胞、星形胶质细胞和上皮样细胞中呈强阳性或阳性反应,被anti-caspase-8和anti-caspase-3抗体染成淡棕色或深棕色。对脑组织蛋白提取物进行Western blot检测,病兔脑组织中TNF-α,caspase-8和caspase-3的表达量明显高于对照兔,差异极显著(P<0.01)。结果表明,兔脑炎原虫可刺激脑组织中的巨噬细胞和上皮样细胞产生大量TNF-α,后者与细胞膜上的TNFR结合,从而激活死亡受体通路,引起胞浆内的caspase家族发生级联反应,导致细胞凋亡。to probe the apoptotic signal transduction pathway caused by Encephalitozoon cuniculi,the cerebral tissues of 40ill rabbits and 10control rabbits were studied particularly by common staining,special staining,immunohistochemistry staining and Western blot in this research.The results showed that the different form of cerebral granulomas and Encephalitozoon cuniculi were detected in cerebral tissues of ill rabbits.TNF-αwas expressed by macrophges and epithelioid cells infected by Encephalitozoon cuniculi.The cells expressed TNF-αwere positive reaction staining to anti-TNF-αantibody.The keys proteins caspase-8and caspase-3of apoptotic signal transduction pathway were presented as strong positive or positive reaction in vascular endothelial cell,astrocytes and epithelioid cells,respectively as light brown or dark brown stained by anti-caspase-8and anti-caspase-3antibody.The cerebral protein extractions were checked by Western blot and shown that proteins expression of TNF-α,caspase-8and caspase-3in sick rabbits were obviously higher than those of control rabbits.The differences were very significant(P<0.01).The conclusions were that macrophage and epithelioid cell stimulated by Encephalitozoon cuniculi could produce lots of TNF-α.While TNF-αcombined with TNFR on cytomembrane,the death receptor pathway was activated,the cascade reaction of caspase was induced in intracytoplasm,and finally led to apoptosis of cerebral cells.

关 键 词：兔脑炎原虫 细胞凋亡信号通路 肿瘤坏死因子 半胱天冬蛋白酶8 半胱天冬蛋白酶3 

分 类 号：S852.72[农业科学—基础兽医学]