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2025年4月3日 星期四
农业生物技术学报  2022, Vol. 30 Issue (4): 676-685    DOI: 10.3969/j.issn.1674-7968.2022.04.006
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
运输应激对山羊免疫器官细胞凋亡及Bcl-2和Bax表达的影响
彭瑞妮1, 刘犇2,3, 余四九1,*
1 甘肃农业大学 动物医学院,兰州 730070;
2 宜春学院 生命科学与资源环境学院,宜春 336000;
3 江西绿科农牧科技有限公司,宜春336000
Effects of Transport Stress on Apoptosis and Expression of Bcl-2 and Bax in the Immune Organs of Goat (Capra hircus)
PENG Rui-Ni1, LIU Ben2,3, YU Si-Jiu1,*
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2 College of Life Science and Resources and Environment, Yichun University, Yichun 336000, China;
3 Jiangxi Lvke Agriculture and Animal Husbandry Technology Co., Ltd., Yichun 336000, China
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摘要 细胞凋亡是维持免疫器官稳态及发挥功能的重要方式,有利于山羊(Capra hircus)在运输应激下维持体内平衡。本研究旨在探讨运输应激对山羊免疫器官细胞凋亡及B细胞淋巴瘤-2 (B-cell lymphoma-2, Bcl-2)、Bcl-2相关X蛋白(B-cell lymphoma-2 associated X protein, Bax)表达的影响。将12头健康赣西公山羊随机分为3组:对照组(不运输)、运输2 h组、运输6 h组。运输完成后采集山羊的免疫器官(脾脏, 肠系膜淋巴结),采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(terminal dexynucleotidyl transferase-mediated dUTP nick end labeling, TUNEL)观察免疫器官细胞凋亡的情况,用免疫组织化学、Western blot和qPCR方法在蛋白和基因水平上进行定位和检测。TUNEL分析结果显示,运输2和6 h组的脾脏和淋巴结细胞凋亡率极显著高于对照组(P<0.01);免疫组化结果显示,运输前后Bax和Bcl-2在脾脏的脾小结及其边缘区和淋巴结的淋巴小结及其周围弥散淋巴组织都有阳性表达,但3组间表达强度不同。Western blot结果显示,脾脏和淋巴结的Bax、Bcl-2蛋白表达量及其比率Bax/Bcl-2在3组间并无统计学差异(P>0.05)。qPCR结果显示,与对照组相比,运输2和6 h组脾脏的BaxBcl-2基因表达量与运输6 h组Bax/Bcl-2的比率均极显著升高(P<0.01),在淋巴结中,运输2 h组BaxBcl-2基因表达量极显著降低(P<0.01),而运输6 h组Bcl-2基因表达量极显著升高(P<0.01),运输2和6 h组Bax/Bcl-2的比率极显著降低(P<0.01)。本研究结果表明,运输应激会导致免疫器官的细胞凋亡增加,且BaxBcl-2基因发生显著变化。本研究为防治运输应激对山羊造成的影响提供了一定的理论依据。
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彭瑞妮
刘犇
余四九
关键词 运输应激脾脏淋巴结B细胞淋巴瘤-2 (Bcl-2)Bcl-2相关X蛋白(Bax)    
Abstract:Apoptosis is an important way to maintain the homeostasis and function of immune organs, which is beneficial to maintain homeostasis in goats (Capra hircus) under transport stress. The study was to investigate the effects of transport stress on apoptosis and the expression of B-cell lymphocytoma-2 (Bcl-2) and Bcl-2 related X protein (Bax) in goats immune organs. Twelve healthy male Ganxi goats were randomly divided into 3 groups: Control group (non transported), transport 2 h group and transport 6 h group. The immune organs (spleen, mesenteric lymph nodes) of goats were collected after transportation. Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to detect apoptosis in immune organs. Immunohistochemistry, Western blot, qPCR methods were used to locate and detect at the protein and gene level. TUNEL analysis showed that the apoptosis rate of spleen and lymph nodes in transport 2 and 6 h were significantly higher than the control group (P<0.01); Immunohistochemical results showed that Bax and Bcl-2 were positively expressed in goats splenic nodules and marginal area of spleen, and lymphatic nodule and surrounding diffuse lymphatic tissue of lymph nodes before and after transportation, but the expression intensity varied among the 3 groups. Western blot results showed that the proteins expression levels of Bax, Bcl-2 and Bax/Bcl-2 ratio were not statistically different among the 3 groups in spleen and lymph nodes (P>0.05). qPCR results showed that, compared with the control group, the mRNA expression levels of Bax and Bcl-2 in transport 2 and 6 h groups and Bax/Bcl-2 ratio in transport 6 h group were significantly increased in spleen. In the lymph nodes, the gene expression levels of Bax and Bcl-2 in transport 2 h group were significantly decreased (P<0.01), and the gene expression of Bcl-2 in 6 h group was significantly increased (P<0.01), Bax/Bcl-2 ratio in transport 2 and 6 h groups was significantly decreased (P<0.01). The results showed that transport stress could lead to increase apoptosis and significant change of the expression of Bax and Bcl-2 in immune organs. The present study provides a theoretical basis for preventing and treating the effects of transport stress in goats.
Key wordsTransport stress    Spleen    Lymph nodes    B-cell lymphocytoma-2 (Bcl-2)    Bcl-2 associated X (Bax)
收稿日期: 2021-04-30     
ZTFLH:  S852.2  
基金资助:国家重点研发项目(2017YFD0502200); 江西现代农业科研协同创新专项(JXXTCX201702)
通讯作者: * sjyu@163.com   
引用本文:   
彭瑞妮, 刘犇, 余四九. 运输应激对山羊免疫器官细胞凋亡及Bcl-2和Bax表达的影响[J]. 农业生物技术学报, 2022, 30(4): 676-685.
PENG Rui-Ni, LIU Ben, YU Si-Jiu. Effects of Transport Stress on Apoptosis and Expression of Bcl-2 and Bax in the Immune Organs of Goat (Capra hircus). 农业生物技术学报, 2022, 30(4): 676-685.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.04.006     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I4/676
 
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