枯草芽胞杆菌肽聚糖介导绵羊瘤胃上皮细胞β-防御素表达的通路研究  

Studies on the signaling pathway of the expression of ORECs β-defensin mediated by Bacillus subtilis peptidoglycan

作　　者：齐盟 胡亮 辛雅明 白绥明 郭铸苇 杨银凤[1,2] QI Meng;HU Liang;XIN Yaming;BAI Suiming;GUO Zhuwei;YANG Yingfeng(College of Veterinary Medicine of Inner Mongolia Agricultural University,Hohhot 010011,China;Key Laboratory of Basic Veterinary Medicine of Inner Mongolia Autonomous Region,Hohhot 010011,China;Bayannur Institute of Agriculture and Animal Husbandry,Bayannur 015000,China;Erdos Animal Disease Prevention and Control Center,Erdos 017000,China;Qingjian County Animal Disease Prevention and Control Center,Shaanxi Province,Yulin 718399,China;Inner Mongolia Animal Disease Prevention and Control Centre/National Centre for Dairy Technology and Innovation,Hohhot 010020,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010011 [2]内蒙古自治区基础兽医学重点试验室,内蒙古呼和浩特010011 [3]巴彦淖尔市农牧业科学研究所,内蒙古巴彦淖尔015000 [4]内蒙古鄂尔多斯市动物疫病预防和控制中心,内蒙古鄂尔多斯017000 [5]陕西省清涧县动物疫病预防控制中心,陕西榆林718399 [6]内蒙古自治区动物疫病预防控制中心/国家乳业技术创新中心,内蒙古呼和浩特010020

出　　处：《畜牧与兽医》2025年第5期43-48,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(32060780);内蒙古自治区教育厅项目(NJZZ21015)。

摘　　要：旨在探究来源于枯草芽胞杆菌细胞壁的肽聚糖诱导绵羊瘤胃上皮细胞(ORECs)β-防御素-1(SBD-1)表达的信号通路情况。使用20μg/mL肽聚糖刺激ORECs 2 h,通过实时荧光定量PCR(RT-qPCR)和蛋白免疫印迹(Western blot)方法,检测SBD-1表达的信号通路因子mRNA和蛋白的表达量;使用特异性抑制剂抑制各信号通路因子,再使用20μg/mL肽聚糖刺激ORECs 2 h,通过RT-qPCR和ELISA方法检测SBD-1 mRNA和蛋白的表达量,确定介导SBD-1表达的主要信号通路。结果:20μg/mL肽聚糖显著提升了Toll样受体2(TLR-2)、髓样分化因子88(MyD88)、细胞外调节蛋白激酶(ERK1/2)、c-Jun氨基末端激酶(JNK)和核因子κB(NF-κB)mRNA和蛋白的表达量(P<0.001,P<0.05),对p38丝裂原活化蛋白激酶(p38 MAPK,P38)的mRNA表达量没有显著影响;使用信号通路抑制剂抑制了ERK1/2、JNK和NF-κB信号通路,极显著降低肽聚糖诱导ORECs中SBD-1的mRNA和蛋白表达量(P<0.001,P<0.01)。结论:枯草芽胞杆菌肽聚糖诱导SBD-1表达由TLR-2-MyD88-NF-κB和TLR-2-MyD88-MAPK(JNK、ERK1/2)共同介导。This study aimed to investigate the signaling pathways through which peptidoglycan derived from the cell wall of Bacillus subtilis induces the expression of sheep β-defensin-1(SBD-1) in ovine rumen epithelial cells(ORECs).ORECs were stimulated with 20 μg/mL peptidoglycan for 2 hours,and the mRNA and protein expression levels of SBD-1-related signaling pathway factors were analyzed using quantitative real-time PCR(RT-qPCR) and Western blot.Then,specific inhibitors were employed to block the individual signaling pathways,followed by stimulation with 20 μg/mL peptidoglycan for 2 hours.Finally,SBD-1 mRNA and protein expression levels were measuredvia RT-qPCR and ELISA to identify the primary signaling pathways involved.The results showed that 20 μg/mL peptidoglycan significantlyupregulated the mRNA and protein expression of TLR-2,MyD88,ERK1/2,JNK,and NF-κB(P<0.001,P<0.05).Inhibition of theERK1/2,JNK,and NF-κB signaling pathways using specific inhibitors markedly reduced peptidoglycan-induced SBD-1 mRNA and proteinexpression in ORECs(P<0.001,P<0.01).These findings indicated that the induction of SBD-1 expression byBacillus subtilispeptidogly-can was mediated through both the TLR-2-MyD88-NF-κB and TLR-2-MyD88-MAPK(JNK,ERK1/2) pathways.

关 键 词：枯草芽胞杆菌肽聚糖 β-防御素-1 绵羊瘤胃上皮细胞 核因子κB 

分 类 号：S826[农业科学—畜牧学]