异体移植炎症因子1通过核因子κB信号促进牛乳腺上皮细胞炎症因子的释放

doi:10.11843/j.issn.0366-6964.2021.03.022

摘要/Abstract

摘要： 奶牛乳腺炎是指在不同理化因素刺激下奶牛乳腺的炎性反应，其严重影响了奶牛养殖业的健康发展。许多细胞因子是炎症调节剂，但与奶牛乳腺炎相关的关键细胞因子还未被鉴定。异体移植炎症因子1（allograft inflammatory factor-1，AIF-1）在免疫调节中扮演重要角色，并在多种炎性疾病中过量表达。因此，本研究探讨了牛AIF-1在乳腺炎中的可能作用。首先，利用ELISA试剂盒检测了乳腺炎牛奶中牛AIF-1的含量，并用RT-PCR方法克隆牛AIF-1基因，然后，用亲和层析纯化牛AIF-1蛋白。用此重组蛋白刺激牛乳腺上皮细胞，ELISA试剂盒检测肿瘤坏死因子α、白细胞介素6和单核细胞趋化蛋白1的分泌，Western blot测定IκBα的磷酸化。结果显示，患乳腺炎奶牛乳中AIF-1的平均含量显著高于健康奶牛，而抗生素治愈后的奶牛乳中AIF-1的含量回落到正常水平。这些结果提示，牛AIF-1是一个分泌型蛋白，可能与乳腺炎相关。为了进一步探索这种关系，本研究克隆了牛AIF-1基因并纯化了牛AIF-1蛋白。此蛋白上调了牛乳腺上皮细胞肿瘤坏死因子α、白细胞介素6和单核细胞趋化蛋白1的分泌，并刺激了IκBα的磷酸化。IκBα磷酸化抑制剂BAY 11-7085阻断了牛AIF-1刺激的炎性细胞因子上调。综上表明，牛AIF-1通过核因子κB信号促进了牛乳腺上皮细胞炎症因子的释放。

关键词: 奶牛乳腺炎, 牛乳腺上皮细胞, 异体移植炎症因子1, 核因子κB

Abstract: Bovine mastitis is the inflammatory response in bovine mammary gland, it is the most prevalent disease in dairy cattle worldwide. Many cytokines are inflammatory mediators. But the key cytokines associated with bovine mastitis have not been elucidated. Allograft inflammatory factor-1 (AIF-1) plays a central role in immune regulation and it is over-expressed in a vast array of inflammatory diseases. Therefore, we explored the possible role of bovine AIF-1 related to bovine mastitis in the present study. First, bovine AIF-1 gene was cloned by RT-PCR and the AIF-1protein was purified by a Ni-NTA spin column. Then, bovine mammary gland epithelial cell MAC-T line was treated with AIF-1 for 48 h. The concentrations of TNF-α, IL-6, AIF-1 and monocyte chemoattractant protein 1 secretion were measured by ELISA assay kits. Finally, phosphorylation of IκBα was detected by Western blot. The results showed that the average concentration of AIF-1 in milks suffering from mastitis was significantly higher than that in the healthy cows, while its value decreased in cows recovered from mastitis. Furthermore, bovine AIF-1 gene was cloned and recombinant bovine AIF-1 protein was prepared. The protein up-regulated TNF-α, IL-6 and monocyte chemoattractant protein 1 secretion from bovine mammary epithelial cells with NF-κB activating, then NF-κB signaling inhibitor BAY 11-7085 abolished the increase of these inflammatory cytokine secretion induced by AIF-1. The present study indicated that AIF-1 induced inflammatory mediator production from bovine mammary epithelial cells via NF-κB signaling.

Key words: bovine mastitis, bovine mammary epithelial cells, allograft inflammatory factor-1, NF-κB

中图分类号:

S857.26

伍迎欢, 杨丹茹, 赵燕英. 异体移植炎症因子1通过核因子κB信号促进牛乳腺上皮细胞炎症因子的释放[J]. 畜牧兽医学报, 2021, 52(3): 782-788.

WU Yinghuan, YANG Danru, ZHAO Yanying. Bovine Allograft Inflammatory Factor-1 (AIF-1) Induces Inflammatory Mediator Secretion from Bovine Mammary Epithelial Cells via NF-κB Signaling[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 782-788.

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