中国兽医杂志2025，Vol.61Issue(2)：18-24,7.DOI:10.20157/j.cnki.zgsyzz.2025.02.003
TAK1调控奶牛乳腺上皮细胞炎症因子和趋化因子的机制
Mechanism of TAK1 in Regulating Inflammatory Factors and Chemokines in Cow Mammary Epithelial Cells
摘要
Abstract
To explore the regulatory mechanism of transforming growth factor β activated kinase 1(TAK1)on inflammatory factors and chemokines in cow mammary epithelial cells(CMECs),this study performed a bioinformatics analysis of the signaling pathways in normal and mastitis-affected CMECs.CMECs were identified via cytokeratin 18(CK-18)immunofluorescence staining and divided into three groups:Control group,Overexpression group,and Inhibition group.The Control group involved normal CMEC culture,the Overexpression group involved CMECs transfected with 2 µg TAK1 overexpression plasmid,and the Inhibition group involved CMECs transfected with 2 µg TAK1 overexpression plasmid followed by treatment with the TAK1 inhibitor 5Z-7-oxozeaenol.Western blot(WB)was used to detect levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),nuclear factor-κB(NF-κB),monocyte chemoattractant protein-1(MCP-1),macrophage inflammatory protein-1α(MIP-1α),and macrophage inflammatory protein-1β(MIP-1β).Aldehyde dehydrogenase(ALDH),a CMEC stem cell marker,was measured via flow cytometry.Bioinformatics analysis revealed activation of cellular immune responses and cell surface receptor signaling pathways in inflamed CMECs,with high-level expression of genes positively regulating inflammation.TNF-α interacted with IL-1β,MIP-1α,MIP-1β,MCP-1,upstream proteins cluster of differentiation 3(CD3)and cluster of differentiation 8(CD8),and chemokine ligand 5(CCL5).WB results showed that compared to the Control group,the Overexpression group exhibited significantly elevated levels of TAK1,inflammatory factors IL-1β,TNF-α and NF-κB,chemokines MIP-1α,MIP-1β,and MCP-1 proteins(P<0.01 or P<0.001),while these protein levels were significantly suppressed in the Inhibition group compared to the Overexpression group(P<0.001).Flow cytometry revealed ALDH-positive cell rates of 6.7%,10.21%,and 4.23%in the Control,Overexpression,and Inhibition groups,respectively,after 48 hours of culture.This study elucidates the regulatory relationship between TAK1 and inflammatory/chemotactic factors in CMECs,providing experimental and theoretical support for treating bovine mastitis.关键词
奶牛乳腺上皮细胞(CMECs)/转化生长因子β激活激酶1(TAK1)/炎症因子/相互作用Key words
cow mammary epithelial cells(CMECs)/transforming growth factor β activated kinase 1(TAK1)/inflammation factor/interaction分类
农业科技引用本文复制引用
陈平,王斐,谢建鹏,张鉴,何振富..TAK1调控奶牛乳腺上皮细胞炎症因子和趋化因子的机制[J].中国兽医杂志,2025,61(2):18-24,7.基金项目
甘肃省科协2023年创新驱动助力工程项目(GXH20230817-9) （GXH20230817-9）
甘肃省农业科学院农业科技自主创新专项博士基金项目(2021GAAS51) （2021GAAS51）
