摘要
羊源CXC趋化因子受体2基因（chemokine C-X-C motif receptor 2,CXCR2）主要表达于炎性细胞表面,在局部炎症反应、肿瘤发生等方面起重要调控作用。为研究CXCR2基因在患临床型乳房炎绵羊乳腺组织中的表达差异及可能的生物学功能,本研究选取正常和患临床型乳房炎的绵羊（Ovis aries）各3只,收集乳腺组织,HE染色观察其组织形态学结构差异,运用实时荧光定量PCR（qRT-PCR）和ELISA技术分别从mRNA和蛋白水平检测两者中CXCR2的表达差异,通过miRbase数据库和miranda软件对绵羊CXCR2基因的靶向miRNAs进行预测,并采用DAVID软件对CXCR2基因的生物功能进行分析。结果表明,与正常组相比,患病组的乳腺组织中结缔组织明显增生,腺泡腔萎缩,腔内含有脱落的上皮细胞并且有大量白细胞浸润;患病组中CXCR2mRNA和蛋白的表达量极显著上调（P〈0.01）;靶向miRNAs预测显示,绵羊CXCR2基因可能受到oar-miR-3956-3p、oar-miR-665-5p、oar-miR-432等miRNAs的调控;GO分析发现CXCR2基因参与构成膜的主要成分、CXC趋化因子受体活性、趋化作用等过程;KEGG通路富集发现CXCR2基因参与细胞因子—受体相互作用、趋化因子信号转导通路和内吞作用信号通路。由此推测,在绵羊乳房炎发生过程中,CXCR2基因可能通过与配体结合,以调控炎性细胞的趋化迁移,进而参与免疫应答反应。本研究为更深入地研究和探讨CXCR2基因在乳房炎发生发展过程中的分子生物学功能提供了研究素材。
The CXCR2 gene is mostly expressed on the surface inflammatory ceils, playing an im- portant regulatory role in local inflammatory reaction, tumorigenesis and so on. The objective of this study was to investigate the expression and potential biological function of the CXCR2 gene during sheep mastitis. Mammary tissues were collected from six lactating Hu sheep（three normal sheep, three clinical mastitic sheep）. HE staining was conducted to observe histomorphology differences in mammary tissues between normal and clinical mastitis,and the expression levels of the CXCR2 mRNA and protein were detected by qRT-PCR and ELISA,respectively. The targeted miRNAs（microRNA）of ovine CXCR2 gene were predicted by miRbase database and miranda soft- ware. Moreover, biological function of the CXCR2 gene was analyzed by DAVID software, t-IE staining showed that mammary tissue sections with clinical mastitis were character- ized by connective tissue hyperplasia, alveolar luminal atrophy,and massive leucocytes and exfolia- ted epithelial cells enriched in acinar chamber compared with normal mammary tissues. The qRT- PCR and ELISA results showed that expression levels of the CXCR2 mRNA and protein in clinical mastitic mammary gland were extremely significantly higher than those in normal tissue （P〈0.01）. The prediction results of targeted miRNAs indicated that ovine CXCR2 gene might be regu- lated by oar-miR-3956-3p,oar-miR-665-5p,oar-miR-432 and so on according to the score. GO an- notation revealed that the CXCR2 gene was involved in integral component of membrane, C-X-C chemokine receptor activity and ehemotaxis. KEGG enrichment analysis indicated that the CXCR2 gene took part in cytokine-eytokine receptor interaction,chemokine signaling pathway and endocy- tosis. Considering together these finding speculate that the CXCR2 may regulate the chemotactic migration of inflammatory cells to participate in inflammatory-immune responses by binding to its ligands. The observation provides basic materials for further research a
作者
宋宜泽
李讨讨
马友记
马轩
王新强
杨海军
安家福
SONG Yi-ze1 ,LI Tao-tao1 ,MA You-ji1,2 ,MA Xuan1 ,WANG Xin-qiang1 ,YANG Hai-jun1 ,AN Jia-fu1(1. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070,China;2. Sheep Breeding Biotechnology Engineering Laboratory of Gansu Province ,Min- qin,Gansu 733300, China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第11期2168-2174,共7页
Chinese Journal of Veterinary Science
基金
甘肃农业大学SRTP资助项目(20170438)
甘肃省科技重大专项计划资助项目(1602NKDH020)
国家肉羊产业技术体系资助项目(CARS-38)
关键词
CXCR2基因
绵羊
乳房炎
表达
chemokine （C-X-C motif） receptor 2 （ CXCR2 ） gene
sheep （ Ovis aries）
mastitis
expres-sion
