肺炎克雷伯氏菌对荷斯坦奶牛乳腺上皮细胞黏附和侵袭的体外研究

畜牧兽医学报 ›› 2008, Vol. 39 ›› Issue (4): 494-498.

肺炎克雷伯氏菌对荷斯坦奶牛乳腺上皮细胞黏附和侵袭的体外研究

王亨，孟霞，邱昌伟，马翀，吴培福，韩超，齐长明

1.中国农业大学动物医学院临床兽医系，北京 100094；2.扬州大学兽医学院，扬州 225009；3 中国农业大学生物学院微生物学与免疫学系，北京 100094；4 北京三元集团，北京 100029

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-04-24 发布日期:2008-04-24

Study on Adherence and Invasion of Holstein Cows Mammary Epithelial Cells by Klebsiella pneumoniae in vitro

WANG Heng, MENG Xia, QIU Chang-wei, MA Chong, WU Pei-fu, HAN Chao, QI Chang-ming

1 Department of Clinical Veterinary, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China; 2 College of Veterinary Medicine， Yangzhou University， Yangzhou 225009, China; 3 Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100094, China; 4 Beijing Sanyuan Lvhe Dairy Group, Beijing 100029, China

Received:1900-01-01 Revised:1900-01-01 Online:2008-04-24 Published:2008-04-24

摘要/Abstract

摘要： 肺炎克雷伯氏菌是否黏附和侵袭乳腺上皮细胞可能是其导致乳房炎的致病机理之一。本试验从荷斯坦奶牛乳腺中分离纯化乳腺上皮细胞，并通过免疫组化和透射电镜观察，证实纯化细胞为乳腺上皮细胞。采用从临床乳房炎病例分离的肺炎克雷伯氏菌攻击原代培养的乳腺上皮细胞，以沙门氏菌和大肠杆菌DH5α为阳性和阴性对照。结果发现，肺炎克雷伯氏菌能够黏附乳腺上皮细胞，且具有时间和剂量依赖性，达到饱和黏附后，不再增加。肺炎克雷伯氏菌能够侵入乳腺上皮细胞，且其活力未受到明显影响，但侵入细菌的数量没有明显的增加，说明一定数量的细菌能够侵入乳腺上皮细胞。肺炎克雷伯氏菌黏附和侵入乳腺上皮细胞并在细胞内保持活力，可能是临床肺炎克雷伯氏菌导致的乳房炎难于治愈的原因之一。

关键词: 肺炎克雷伯氏菌, 黏附, 侵袭, 牛乳房炎

Abstract: Adherence and invasion of Klebsiella pneumoniae to bovine mammary epithelial cells can be the potentially pathogenic mechanisms. In the present study, the cultured bovine mamary epithelial cells was applied to detect the adherence and invasion of Klebsiella pneumoniae. The cells were determined by immunohistochemistry with anti-cytokeratin antibody and transmission electron microscopy. Positive immuno-reaction and desmosomes were observed respectively. Klebsiella pneumoniae, originating from clinical bovine mastitis cases, Salmonella typhimurium and Escherichia coli DH5α were tested for their ability of adherence and invasion to bovine mammary epithelial cells in vitro Results showed that Klebsiella pneumoniae adhered to bovine mammary epithelial cells in dose and time dependent until saturation, and invaded the cells without attenuation. In addition, no significant influence on the bacteria viability in cells was observed during the experimental time. The results of the present paper may partially contribute to an understanding of the pathogenesis of Klebsiella pneumoniae mastitis by surviving in the cells to protect themselves from antimicrobial drugs and host defense system.

Key words: Klebsiella pneumoniae, adherence, invasion, bovine mastitis

王亨;孟霞;邱昌伟;马翀;吴培福;韩超;齐长明. 肺炎克雷伯氏菌对荷斯坦奶牛乳腺上皮细胞黏附和侵袭的体外研究[J]. 畜牧兽医学报, 2008, 39(4): 494-498.

WANG Heng;MENG Xia;QIU Chang-wei;MA Chong;WU Pei-fu;HAN Chao;QI Chang-ming. Study on Adherence and Invasion of Holstein Cows Mammary Epithelial Cells by Klebsiella pneumoniae in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2008, 39(4): 494-498.

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