绵羊CD14基因在临床型乳腺炎乳腺组织中的表达  

Expression of sheep CD14 in mammary tissue with clinical mastitis

作　　者：范梓文 尹德恩 满永恒 郭磊 马友记[1,2] FAN Ziwen;YIN Deen;MAN Yongheng;GUO Lei;MA Youji(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation,Lanzhou 730070,China;Animal Husbandry and Veterinary Technical Service Center of Yongchang County,Jinchang,Gansu 737200,China)

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070 [2]甘肃省动物生殖生理与繁殖调控重点实验室,甘肃兰州730070 [3]甘肃省金昌市永昌县畜牧兽医技术服务中心,甘肃金昌737200

出　　处：《中国兽医学报》2023年第8期1696-1702,共7页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2021YFD1100502)。

摘　　要：白细胞分化抗原14(cluster of differentiation antigen 14,CD14)是由成熟的单核巨噬细胞分泌的多态类炎症细胞因子,在细胞激活、免疫反应调节和机体炎症反应过程中起到关键作用。到目前为止,关于CD14基因在绵羊乳腺炎乳腺组织发生过程的研究仍处于空白阶段。为分析CD14基因的序列以及在临床型乳腺炎乳腺组织中的表达特征,本研究基于NCBI网站已公布的绵羊CD14 CDS序列,对该基因生物信息学进行分析。使用qRT-PCR、Western blot与免疫组织化学染色的方法检测CD14 mRNA及其蛋白在健康和临床型乳腺炎绵羊乳腺组织中的表达与分布情况。生物信息学分析结果显示绵羊CD14为疏水性不稳定蛋白、分泌蛋白,在进化过程中CD14基因高度保守趋于稳定。qRT-PCR与Western blot结果显示,临床型乳腺炎乳腺组织中CD14 mRNA显著高于健康组(P<0.05),CD14蛋白的相对表达量极显著高于健康组(P<0.01)。免疫组织化学检测结果显示在健康绵羊乳腺组织细胞中的CD14蛋白主要存在于基质细胞中,而在临床型乳腺炎乳腺组织中于乳腺上皮细胞和基质细胞中均强烈分布。这可能是由于CD14基因表达的上调,导致患临床型乳腺炎乳腺组织中基质细胞与乳腺上皮细胞中炎症反应加剧。为进一步研究CD14基因在绵羊乳腺炎发病机制中的调控作用提供了基础信息。Cluster of differentiation antigen 14(CD14)is a polymorphic class of inflammatory cytokines secreted by mature monocytes and plays a key role in cell activation,regulation ofthe immune response and the inflammatory response of the body.To date,studies on the CD14 gene in the mammary gland histogenesis of mastitis in sheep are still in a gap.To characterize the sequence of CD14 gene and its expression in the mammary tissue of clinical mastitis,this study was performed to analyze the bioinformatics of this gene based on the published sheep CD14 CDS sequence on the NCBI website.The test used qRT-PCR,Western blot and immunohistochemical staining to detect the expression and distribution of CD14 mRNA and its protein in mammary tissues of healthy and clinically mastitis sheep.The results of bioinformatics analysis showed that CD14 was a hydrophobic,unstable protein,a secretory protein,and the CD14 gene was highly conserved and stabilized during the evolutionary process.qRT-PCR and protein immunoblotting showed that CD14mRNA was significantly(P<0.05)higher in clinical mastitis mammary tissue than in the healthy group,and the relative expression of CD14protein was extremely high(P<0.01).The relative expression of CD14protein was higher than that of the healthy group.The immunohistochemical results showed that CD14protein was mainly present in stromal cells in healthy sheep mammary tissue,while it was strongly distributed in both mammary epithelial cells and stromal cells in clinical mastitis mammary tissue.This may be due to the upregulation of CD14gene expression,which leads to an increased inflammatory response in stromal cells and mammary epithelial cells in mammary tissue with clinical mastitis.This provides basic information for further studies on the regulatory role of the CD14gene in the pathogenesis of mastitis in sheep.

关 键 词：绵羊 白细胞分化抗原14(CD14) 乳腺炎 表达 定位 

分 类 号：S858.26[农业科学—临床兽医学]