摘要
为了探究禽网状内皮组织增生病病毒(reticuloendotheliosis virus,REV)感染无特殊病原体(specific pathogen free,SPF)雏鸡后,SPF雏鸡胸腺细胞凋亡的动态变化及细胞凋亡与B细胞淋巴瘤/白血病-2基因(B cell lymphoma/lewkmia-2,BCL-2)mRNA和蛋白表达的相互关系,将60只1日龄SPF雏鸡随机分为对照组和REV感染组,在病毒感染后1,7,14,21,28,42 d分别采集上述2组雏鸡的胸腺,应用TUNEL法测定胸腺细胞的原位凋亡变化,用间接ELISA法和免疫组织化学法分别测定BCL-2蛋白含量及阳性细胞数量,用qRT-PCR检测BCL-2的mRNA表达。结果发现,与对照组雏鸡比较,REV感染雏鸡的胸腺细胞凋亡数量在整个试验期间均出现增多,且在病毒感染后21,28 d明显增多(P<0.05或P<0.01)。与此同时,BCL-2的mRNA表达、蛋白含量以及BCL-2阳性细胞数量均增多,并在病毒感染后21,28 d显著增多(P<0.05或P<0.01)。上述结果表明,REV感染的SPF雏鸡胸腺细胞BCL-2 mRNA表达和蛋白含量的增加与病毒感染导致的胸腺细胞凋亡密切相关,胸腺细胞凋亡的增加是REV感染所致雏鸡细胞免疫功能低下的直接因素。
The present study aimed to determine the dynamic changes of apoptotic thymic cells,and the correlation between apoptosis and BCL-2 mRNA and protein expression in thymus of experimentally infected and uninfected SPF chicks.Sixty 1-day-old chicks were divided as group control and group infection randomly,and their thymus were collected at 1,7,14,21 and 28 d post infection with REV.TUNEL method was performed to detect the changes of apoptotic thymic cells in situ.Indirect ELISA and IHC were also performed to detect BCL-2 protein and BCL-2 positive cells separately,and the BCL-2 mRNA was analyzed by qRT-PCR.The results showed that REV-infected SPF chicks got an increase in apoptotic thymic cells in varying degree,especially from 21 to 28 dpi(P<0.05 or P<0.01).However,Meanwhile,the mRNA expression and protein content of BCL-2 as well as the number of BCL-2 positive cells were significantly increased at 21 to 28 dpi(P<0.05 or P<0.01).These results suggest that the increase in BCL-2 mRNA and protein in thymocytes of REV-infected SPF chicks is closely related to apoptotic thymic cells caused by viral infection.The increase in thymocyte apoptosis due to REV infection directly lead to the low cellular immune function in chicks.
作者
吴文杰
赵晨辉
边洋
高雪丽
刘超男
吕晓萍
顾先哲
郑世民
WU Wenjie;ZHAO Chenhui;BIAN Yang;GAO Xueli;LIU Chaonan;LYU Xiaoping;GU Xianzhe;ZHENG Shimin(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2021年第12期2318-2324,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31472169)。