Acta Veterinaria et Zootechnica Sinica ›› 2022, Vol. 53 ›› Issue (8): 2763-2772.doi: 10.11843/j.issn.0366-6964.2022.08.033

• CLINICAL VETERINARY MEDICINE • Previous Articles     Next Articles

Construction of Dairy Cow Mastitis Model and Analysis of mRNA Transcription Level of Inflammation Related Cytokine Genes

LUORENG Zhuoma1,2, WANG Jinpeng1,2, JIAO Peng1,2, LI Yanxia1,2, DONG Yiwen1,2, WEI Dawei1,2, WNAG Xingping1,2*   

  1. 1. School of Agricultural, Ningxia University, Yinchuan 750021, China;
    2. Key Laboratory of Ruminant Molecular Cell Breeding, Ningxia Hui Autonomous Region, Yinchuan 750021, China
  • Received:2021-11-25 Online:2022-08-23 Published:2022-08-23

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Abstract: The study aimed to investigate the mRNA transcription levels of inflammation related cytokine genes in mammary tissues of dairy cows with S. aureus or E. coli type mastitis. S. aureus or E. coli (105 CFU·mL-1) were respectively injected into the cow's udder through its duct. On the 7th day of infection, the mammary tissues were collected by aseptic operation in vivo. Then the mastitis models were identified by HE staining and immunofluorescence of mammary tissues. qPCR was used to detect the mRNA transcription levels of nine genes in cow mammary tissues of two induced groups and control group, including chemokine family (CCL2, CCL8, CXCR1, CXCL2 and CXCL13), complement factor (CFI and CFB), autophagy factor DEPP1 and interleukin receptor IL21R. Compared with the control group, the protein expressions of the key molecules (TLR4, NF-κB and TNF α) in TLR4/NF-κB signaling pathway related inflammation were extremely significantly up-regulated in the mammary tissues of the two induced groups (P<0.01), indicating that two types of mastitis in vivo models were successfully constructed combined with the results of HE staining. The results of mRNA transcription levels detection showed that, compared with the control group, the mRNA transcription levels of seven genes (CCL2, CCL8, CXCR1, CXCL2, CFI, CFB and IL21R) were extremely significantly up-regulated in mammary tissues of the two induced groups (P<0.001), the mRNA transcription levels of CXCL13 were extremely significantly up-regulated only in mammary tissues of the S. aureus induced group (P<0.01), while the expression of DEPP1 was extremely significantly down-regulated in mammary tissues of the both induced groups (P<0.01). In addition, the mRNA transcription levels of five genes including CCL2, CCL8, CXCR1, CFI and IL21R in E. coli induced group were significantly higher than those in S. aureus induced group (P<0.01). S. aureus or E. coli infection can cause serious clinical mastitis symptoms in dairy cows, and promote the abnormal expression of the above inflammation related genes in mammary tissues, to response to the occurrence and development process of mammary inflammation. The mRNA transcription levels of five genes including chemokine CCL2 in E. coli induced group were significantly higher than that in S. aureus induced group, which explained that E. coli often caused acute mastitis, while S. aureus caused chronic mastitis. The above results will provide the reference for further research on the molecular regulation mechanism of different type mastitis in dairy cow.

Key words: cow, mastitis, gene expression, inflammation

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