Dectin-2和TLR-2在酿酒酵母甘露聚糖诱导SBD-1表达过程中的作用  

The role of Dectin-2 and TLR-2 in the expression of SBD-1 induced by Saccharomyces cerevisiae mannan

作　　者：金鑫[1,2] 张曼[1,2] 王云鹤[1,2] 魏方 温婧怡 杨银凤[1,2] JIN Xin;ZHANG Man;WANG Yun-he;WEI Fang;WEN Jing-yi;YANG Yin-feng(College of Veterinary,Inner Mongolia Agricultural University,Hohhot 010018,China;Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease,Ministry of Agriculture,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2020年第1期122-128,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31560682);内蒙古自治区研究生教育创新计划资助项目(B20171012921)

摘　　要：为了探索膜受体Dectin-2和TLR-2在酿酒酵母甘露聚糖诱导绵羊瘤胃上皮细胞(ovine ruminal epithelial cells,ORECs)β-防御素-1(sheepβ-defensin-1,SBD-1)表达过程中的作用。本研究首先利用免疫组化、RT-PCR和Western blot方法确定Dectin-2是否在ORECs内表达;然后采用qPCR和Western blot方法检测甘露聚糖刺激ORECs后Dectin-2和TLR-2的表达变化;最后用qPCR和ELISA方法检测不同质量浓度(0.1,1.0,10.0 mg/L)的Dectin-2和TLR-2特异性封闭抗体对甘露聚糖诱导SBD-1表达的影响。结果显示:ORECs内表达Dectin-2,且甘露聚糖刺激ORECs后Dectin-2和TLR-2的表达量均显著增加(P<0.01);同时甘露聚糖可以诱导SBD-1的表达(P<0.01),但是该表达过程可以被不同质量浓度的Dectin-2封闭抗体和TLR-2封闭抗体所抑制(P<0.05),且随着抗体浓度的增加,这种抑制作用越明显。此外,在封闭抗体质量浓度相同时,Dectin-2封闭抗体比TLR-2封闭抗体抑制SBD-1表达的作用更明显(P<0.05)。结果表明,酿酒酵母甘露聚糖诱导SBD-1的表达与Dectin-2和TLR-2受体有关,但是Dectin-2在此过程中发挥更主要的作用。In order to explore the role of membrane receptors Dectin-2 and TLR-2 in the expression of sheep β-defensin-1(SBD-1) in ovine ruminal epithelial cells(ORECs) induced by Saccharomyces cerevisiae mannan.In this study,immunohistochemistry,RT-PCR and Western blot were used to determine whether Dectin-2 was expressed in ORECs.Then qPCR and Western blot were used to detect the expression changes of Dectin-2 and TLR-2 after mannan stimulation of ORECs.Finally,qPCR and ELISA were used to detect the effects of different concentrations(0.1,1.0,10.0 mg/L) of Dectin-2 and TLR-2 specific blocking antibodies on the expression of SBD-1 induced by mannan.The results showed that Dectin-2 was expressed in ORECs,and the expression of Dectin-2 and TLR-2 were significantly increased after mannan stimulation of ORECs(P<0.01).At the same time,mannan could induce the expression of SBD-1(P<0.01),but the expression could be inhibited by different concentrations of Dectin-2 blocking antibody and TLR-2 blocking antibody(P<0.05),and the inhibition is more obvious with the increase of antibody concentration.In addition,when the blocking antibody concentration was the same,Dectin-2 blocking antibody was more effective than TLR-2 blocking antibody in inhibiting SBD-1 expression(P<0.05).The results showed that the expression of SBD-1 induced by Saccharomyces cerevisiae mannan was related to Dectin-2 and TLR-2 receptors,but Dectin-2 played a more important role in this process.

关 键 词：甘露聚糖 β-防御素-1 绵羊瘤胃上皮细胞 Dectin-2 TLR-2 

分 类 号：S852[农业科学—基础兽医学]