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当前位置: 首页 > 文章 > CBL蛋白表达对H9N2亚型禽流感病毒细胞中复制及感染细胞凋亡的影响 畜牧与兽医 2019 (5) 108-113
Position: Home > Articles > CBL蛋白表达对H9N2亚型禽流感病毒细胞中复制及感染细胞凋亡的影响 Animal Husbandry & Veterinary Medicine 2019 (5) 108-113

CBL蛋白表达对H9N2亚型禽流感病毒细胞中复制及感染细胞凋亡的影响

作  者：
郑阳;郝珊珊;张则;蔡佳希;宗嫚嫚;余远楠;田晓宁;冯秀丽
单  位：
南京农业大学动物医学院/农业部动物细菌学重点实验室/教育部"动物健康与食品安全"国际合作联合实验室
关键词：
CBL;真核表达;H9N2亚型病毒;细胞凋亡
摘  要：
CBL蛋白是一种E3泛素连接酶,在免疫调控、信号传导调节和对多种刺激的反应中起重要作用.本研究的主要目的是探讨真核表达的CBL蛋白对H9N2禽流感病毒增殖及感染细胞的凋亡情况的影响.荧光显微镜下观察真核表达重组载体EGFP-CBL在MDCK细胞内正常表达,qPCR结果证实禽流感病毒H9N2亚型感染的转染EGFP-CBL重组载体的MDCK细胞内病毒滴度明显少;Western blot结果证实禽流感病毒H9N2亚型感染的转染EGFP-CBL重组载体的MDCK细胞36 h,其凋亡蛋白Bcl-2/Bax比值较大;流式分析结果显示转染EGFP-CBL重组载体的MDCK的早期凋亡细胞的比例少,暗示CBL蛋白能够一定程度上抑制MDCK细胞的凋亡.该结果表明CBL蛋白能在一定程度上抑制H9N2禽流感病毒在MDCK细胞的扩增,并且抑制细胞的凋亡,为深入研究CBL蛋白抗病毒细胞机制提供了重要的试验依据.
作  者：
ZHENG Yang;HAO Shanshan;ZHANG Ze;CAI Jiaxi;ZONG Manman;YU Yuannan;TIAN Xiaoning;FENG Xiuli;College of Veterinary Medicine, Nanjing Agricultural University/Key Laboratory of Animal Microbiology of China's Ministry of Agriculture/MOE Joint International Research Laboratory of Animal Health and Food Safety;
单  位：
ZHENG Yang%HAO Shanshan%ZHANG Ze%CAI Jiaxi%ZONG Manman%YU Yuannan%TIAN Xiaoning%FENG Xiuli%College of Veterinary Medicine, Nanjing Agricultural University/Key Laboratory of Animal Microbiology of China's Ministry of Agriculture/MOE Joint International Research Laboratory of Animal Health and Food Safety
关键词：
CBL;;eukaryotic expression;;H9N2AIV;;apoptosis
摘  要：
Casitas B-lineage lymphoma(CBL) is an E3 ubiquitin ligase, known to play an essential role in the regulation of signal transduction, immunology and some stimulating responses. This paper was to investigate the effect of eukaryotic expression of CBL protein on the proliferation and apoptosis of MDCK cells infected with the H9N2 avian influenza virus. The results of fluorescence microscopy showed that the recombinant EGFP-CBL was expressed in the MDCK cells, and the red fluorescence stained AIV viral particles in EGFP-CBL recombinant protein group were less than those of the EGFP vector plus AIV group; and the results of qPCR confirmed that the viral titers of the MDCK cells transfected with the recombinant EGFP-CBL vector were significantly lower than those of the EGFP vector plus AIV group. Also, Western Blot results confirmed that the expression ratio of the apoptotic protein Bcl-2/Bax in the MDCK cells transfected with recombinant EGFP-CBL vector and with H9N2 AIV was higher than that of the EGFP vector plus AIV group at 36 h of post incubation. Furthermore, the FCM results showed that the percentage of the early apoptotic cells of MDCK transfected with EGFP-CBL recombinant vector and with H9N2 AIV was lowered, compared with that of the EGFP vector plus AIV group; which suggested that the CBL protein might inhibit the apoptosis of MDCK after infection with AIV. The present study indicated that CBL proteins inhibited the replication of H9N2 AIV in MDCK cells and the apoptosis of MDCK cells after infection with AIV; which provided an important experimental basis for further research on the antiviral mechanism of CBL proteins.

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