肺炎支原体感染猪模型的建立及相关指标分析
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摘要
旨在建立肺炎支原体感染猪模型。选取姜曲海猪和杜洛克猪的杂交一代为试验动物,随机分成感染组和对照组,人工接种猪肺炎支原体,试验期40 d,期间观察猪群临床症状,称体质量,检测血清抗体和IgG水平,屠宰后检查肺组织病变,并采用qRT-PCR检测免疫相关分子基因的表达。结果显示,试验16 d后感染组出现支原体肺炎临床症状;试验30 d后,感染组的体质量极显著低于对照组;试验20 d后,感染组血清肺炎支原体抗体阳性,IgG质量浓度极显著高于对照组;试验40 d时屠宰,病理检查发现,感染组表现严重的肺组织病变,脾和肺组织的 IL-1β、 TLR4、 IL-4、 IL-2基因表达显著高于对照组。结果表明,感染组表现出典型支原体肺炎临床表现和病理变化,成功建立肺炎支原体感染猪模型。
In order to establish an animal model for Mycoplasma hyopneumonia(Mhp) infection, crossbred offspring of Jiangquhai pig and Duroc pig were used as experimental materials,the experimental pigs were divided into infection group and control group randomly and the infection group was inoculated with Mhp. The experiment lasted 40 days, clinical feature,body mass change, antibody and IgG production, pulmonary lesion, immunological molecules expression were detected for evaluating the impact of Mhp infection. The experiment results showed the infected pigs appeared clinical symptoms of Mhp at the 16 th-day post inoculation(dpi). Starting 30 th dpi, the average body mass of infected pigs was significantly lower than that of control pigs; starting 20 th dpi, the infected pigs were detected positive Mhp antibody titers in the serum and showed very significantly higher levels of IgG compared with control pigs. At the 40 th dpi, all experimental pigs were slaughtered and dissected; the infected pigs showed serious Mhp pulmonary lesions. In spleen and lung tissue of infected pigs, we observed significantly higher levels of IL-1β, TLR4, IL-4, IL-2 gene expression. The results showed that the infected pigs showed Mhp-typical clinical symptoms and pathologic changes, and successfully established a pig model of Mhp infection.
In order to establish an animal model for Mycoplasma hyopneumonia(Mhp) infection, crossbred offspring of Jiangquhai pig and Duroc pig were used as experimental materials,the experimental pigs were divided into infection group and control group randomly and the infection group was inoculated with Mhp. The experiment lasted 40 days, clinical feature,body mass change, antibody and IgG production, pulmonary lesion, immunological molecules expression were detected for evaluating the impact of Mhp infection. The experiment results showed the infected pigs appeared clinical symptoms of Mhp at the 16 th-day post inoculation(dpi). Starting 30 th dpi, the average body mass of infected pigs was significantly lower than that of control pigs; starting 20 th dpi, the infected pigs were detected positive Mhp antibody titers in the serum and showed very significantly higher levels of IgG compared with control pigs. At the 40 th dpi, all experimental pigs were slaughtered and dissected; the infected pigs showed serious Mhp pulmonary lesions. In spleen and lung tissue of infected pigs, we observed significantly higher levels of IL-1β, TLR4, IL-4, IL-2 gene expression. The results showed that the infected pigs showed Mhp-typical clinical symptoms and pathologic changes, and successfully established a pig model of Mhp infection.
引文
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[2] 倪博,白昀,甘源,等.猪肺炎支原体强弱毒株感染后呼吸道特异性sIgA抗体分泌规律差异性 [J].中国兽医学报,2018,38(7):1307-1310.NI B,BAI Y,GAN Y,et al.The difference in sIgA antibody production of swine respiratory tract after the virulent and avirulent strains of Mycoplasma hyopneumoniae infection [J].Chinese Journal of Veterinary Science,2018,38(7):1307-1310.
[3] 邢福珊,王韦华,张彦明,等.猪支原体肺炎的流行病学调查及病理学诊断 [J].西北农业学报,2009,18(5):67-70.XING F SH,WANG W H,ZHANG Y M,et al.Eepidemiological investigation and pathological histology observation of mycoplasmal pneumonia of swine[J].Acta Agriculturae Boreali-occidentalis Sinica,2009,18(5):67-70.
[4] BORJIGIN L,SHIMAZU T,KATAYAMA Y,et al.Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity [J].Animal Science Journal,2016,87(5):638-645.
[5] BERGMAN I M,ROSENGREN J K,EDMAN K,et al.European wild boars and domestic pigs display different polymorphic patterns in the Toll-like receptor (TLR) 1, TLR2,and TLR6 genes [J].Immunogenetics,2010,62(1):49-58.
[6] 方晓敏,刘筱,孟翠,等.猪Toll样受体基因的变异及其与支原体肺炎感染的关系 [J].华北农学报,2012,27(2):6-11.FANG X M,LIU X,MENG C,et al.Genetic variation of toll-like receptors and Mycoplasma pneumoniae infection in pigs[J].Acta Agriculturae Boreali-Sinica,2012,27(2):6-11.
[7] 邵国青,刘茂军,孙佩元,等.猪气喘病实验猪模型的建立 [J].微生物与感染,2007,2(4):215-218.SHAO G Q,LIU M J,SUN P Y.et al.The establishment of an experimental swine model of swine mycoplasma pneumonia[J].Journal of Microbes and Infection,2007,2(4):215-218.
[8] OSTANELLO F,DOTTORI M,GUSMARA C,et al.Pneumonia disease assessment using a slaughterhouse lung-scoring method [J].Journal of Veterinary Medicine Series A-physiology Pathology Clinical Medicine,2007,54(2):70-75.
[9] 方晓敏,赵为民,付言峰,等.猪支原体肺炎发生的品种敏感差异及分子基础 [J].中国农业科学,2015,48(14):2839-2847.FANG X M,ZHAO W M,FU Y F.et al.Difference in susceptibility to Mycoplasma pneumoniae among various pig breeds and its molecular genetic basis[J].Scientia Agricultura Sinica,2015,48(14):2839-2847.
[10] 李彦伟,刘茂军,武昱孜,等.猪肺炎支原体致病机理研究进展 [J].动物医学进展,2013,34(8):84-88.LI Y W,LIU M J,WU Y Z,et al.Advance in pathogenic mechanism of Mycoplasma hyopneumoniae[J].Progress in Veterinary Medicine,2013,34(8):84-88.
[11] 杜德燕,方鹏飞,朱冬梅,等.猪支原体肺炎病变对体重的影响 [J].中国兽医杂志,2018,54(6):61-63.DU D Y,FANG P F,ZHU D M,et al.Effect of Mycoplasma pneumonia inflammatorye lesion change on body weight [J].Chinese Journal of Veterinary Medicine,2018,54(6):61-63.
[12] SIBILA M,MENTABERRE G,BOADELLA M,et al.Serological,pathological and polymerase chain reaction studies on Mycoplasma hyopneumoniae infection in the wild boar [J].Veterinary Microbiology,2010,144(1):214-218.
[13] JACOBS E,STUHLERT A,DREWS M,et al.Host reactions to Mycoplasma pneumoniae infections in guinea-pigs preimmunized systemically with the adhesin of this pathogen [J].Microbial Pathogenesis,1988,5(4):259-265.
[14] 邓均华,熊焰.猪肺炎支原体与猪支气管败血波氏杆菌在猪体液免疫和生长性能上的协同作用 [J].中国预防兽医学报,2005,27(3):223-226.DENG J H,XIONG Y.Synergism of Mycoplasma hyopneumoniae and Boidatella bronchisepticai in humoral response and growth performance of pigs [J].Chinese Journal of Preventive Veterinary Medicine,2005,27(3):223-226.
[15] 石君霞,刘玉兰,鲁晶,等.脂多糖对断奶仔猪外周血免疫细胞和免疫器官中PPARγ mRNA表达水平的影响 [J].畜牧兽医学报,2008,39(5):608-613.SHI J X,LIU Y L,LU J,et al.Effects of lipopolysaccharide challenge on PPARγ mRNA expression in immune cells and immune organs of weanling piglet [J].Acta Veterinaria Et Zootechnica Sinica,2008,39(5):608-613.
[16] MEDVEDEV A E.Toll-like receptor polymorphisms,inflammatory and infectious diseases,allergies,and cancer [J].Journal of Interferon & Cytokine Research,2013,33(9):467-484.
[17] THADA S,VALLURI V L,GADDAM S L.Influence of Toll-like receptor gene polymorphisms to tuberculosis susceptibility in humans [J].Scandinavian Journal of Immunology,2013,78(3):221-229.
[18] HUNT K A,ZHERNAKOVA A,TURNER G,et al.Novel celiac disease genetic determinants related to the immune response [J].Nature Genetics,2008,40(4):395-402.
[19] RAMíREZ A S,ROSAS A,HERNáNDEZ-BERIAIN J A,et al.Relationship between rheumatoid arthritis and Mycoplasma pneumoniae:a case-control study [J].Rheumatology,2005,44(7):912-914.
[20] 卿素珠,张靖飞,张为民,等.抗病毒合剂对机体免疫器官的影响 [J].西北农业学报,2002,11( 2):18-20.QING S ZH,ZHANG J F,ZHANG W M,et al.Influence of anti-virus mixture on immune organs of body [J].Acta Agriculturae Boreali-occidentalis Sinica,2002,11( 2):18-20.
[21] PIETSCH K,EHLERS S,JACOBS E.Cytokine gene expression in the lungs of BALB/c mice during primary and secondary intranasal infection with Mycoplasma pneumoniae [J].Microbiology,1994,140(8):2043-2048.
[22] SHI C S,SHENDEROV K,HUANG N N,et al.Activation of autophagy by inflammatory signals limits IL-1β production by targeting ubiquitinated inflammasomes for destruction [J].Nature Immunology,2012,13(3):255-263.