化脓隐秘杆菌对奶牛脾脏组织细胞凋亡的影响  

Effect of Trueperella pyogenes on apoptosis of spleen tissue of dairy cows

作　　者：范春玲[1] 颜廷富 张林雁 王洋[3] 周玉龙[1] 孙斌[1] 郭东华[1] 原冬伟[1] 赵明迪 吴海燕[1] 刘琳珊[1] FAN Chun-ling;YAN Ting-fu;ZHANG Lin-yan;WANG Yang;ZHOU Yu-long;SUN Bin;GUO Dong-hua;YUAN Dong-wei;ZHAO Ming-di;WU Hai-yan;LIU Lin-shan(College of Animal Medicine,Heilongjiang Bayi Agriculture University,Daqing 163319,Heilongjiang Province,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江八一农垦大学人文学院,黑龙江大庆163319 [3]黑龙江八一农垦大学校医院,黑龙江大庆163319

出　　处：《中国生物制品学杂志》2022年第1期45-49,共5页Chinese Journal of Biologicals

基　　金：黑龙江省教育厅科学技术研究项目(12541582);黑龙江省农垦总局重点科研计划项目(HKKYZD190312)。

摘　　要：目的观察化脓隐秘杆菌自然感染奶牛脾脏组织的病理学变化,并检测组织细胞中Caspase-3、Caspase-8、Caspase-9、Bax和Bcl-2的表达,以确定化脓隐秘杆菌感染对奶牛脾脏组织细胞凋亡的影响。方法筛检2头化脓隐秘杆菌阴性健康奶牛(对照)和5头自然感染化脓隐秘杆菌的奶牛,收集脾脏组织样本,采用本课题组国家发明专利方法,进行石蜡制片和HE染色,观察脾脏组织病理学变化;采用免疫组织化学方法与本课题组国家发明专利方法相结合,检测奶牛脾脏组织中与细胞正负凋亡相关的调节基因Bax和Bcl-2及凋亡通路蛋白Caspase-3、Caspase-8和Caspase-9的表达情况。结果肉眼观察可见化脓隐秘杆菌自然感染奶牛脾脏肿大、柔软,有出血;病理组织学观察可见脾脏组织出血,水肿,淋巴细胞数量减少,有坏死;脾脏淋巴细胞Caspase-3、Caspase-9和Bax表达阳性,与对照组相比,差异有统计学意义(P <0.05),少量淋巴细胞Caspase-8和Bcl-2表达阳性,与对照组相比,差异无统计学意义(P> 0.05)。结论化脓隐秘杆菌能够引起奶牛脾脏淋巴细胞发生坏死和凋亡;可通过调节脾脏中淋巴细胞Bcl-2家族的活性,引起其Bax和Bcl-2比率改变,增加淋巴细胞线粒体内外膜通透性,导致与凋亡相关因子的释放,激活Caspase-9,进一步激活Caspase-3,从而引起脾脏中淋巴细胞发生凋亡。Objective To observe the pathological change of spleen tissue of dairy cows naturally infected with Trueperella pyogenes and the expressions of Caspase-3,Caspase-8,Caspase-9,Bax and Bcl-2 in splenocytes so as to determine the effect of T. pyogenes infection on apoptosis of spleen tissue of cows. Methods Specimens of spleen tissue were collected from two healthy dairy cows negative for T. pyogenes(control) and five dairy cows infected naturally with T. pyogenes,prepared into paraffin sections by the method developed by the authors,with national invention patent,and subjected to HE staining to observe the pathological change. The expressions of regulatory genes Bax and Bcl-2 associated with positive and negative apoptosis respectively and apoptosis pathway-associated proteins Caspase-3,Caspase-8 and Caspase-9 were determined by immunohistochemical assay combined with the developed method with national patent.Results The spleens of dairy cows naturally infected with T. pyogenes were enlarged and soft with bleeding. Histopathologic observation showed hemorrhage,interstitial edema as well as decrease in number and necrosis of lymphocytes.Caspase-3,Caspase-9 and Bax were expressed positively in the lymphocytes of spleens of infected cows,while the number of positive lymphocytes were significantly larger than that in control group(P < 0. 05). However,a small number of lymphocytes in infected cows were positive for Caspase-8 and Bcl-2 expressions,which showed no significant difference with that in control group(P > 0. 05). Conclusion T. pyogenes may cause the necrosis and apoptosis of lymphocytes in spleens of diary cows. By regulating the activity of Bcl-2 family of lymphocytes in spleen,T. pyogenes may change the ratio of Bax to Bcl-2,increase the permeability of the inner and outer membrane of mitochondria,lead to the release of apoptosis-related factors,activate Caspase-9,and further activate Caspase-3,finally cause the apoptosis of lymphocytes in spleen.

关 键 词：牛 化脓隐秘杆菌 脾脏 凋亡 信号通路 

分 类 号：S852.3[农业科学—基础兽医学]