畜牧兽医学报2017，Vol.48Issue(3)：561-569,9.DOI:10.11843/j.issn.0366-6964.2017.03.021
酿酒酵母菌诱导绵羊瘤胃上皮细胞SBD-1表达的信号通路初步研究
Preliminary Study on the Signaling Pathways of SBD-1 Expression in Sheep Rumen Epithelial Cells Induced by Saccharomyces cerevisiae
摘要
Abstract
In order to explore the regulation mechanism of probiotic Saccharomyces cerevisiae inducing Sheep Beta-Defensin-1 (SBD-1) gene expression in sheep rumen epithelial cells (RECs),we studied the NF-κB and MAPKs signal pathways.Firstly,the RECs of sheep were cultured successfully as in vitro experimental model,and then selected the concentration and incubation time of S.cerevisiae,inducing SBD-1 expression highest,to preliminarily study the subsequent signaling pathways.Real-time fluorescence quantitative PCR (RT-qPCR) was conducted to determine mRNA expressive variation of the toll like receptor 2 (TLR2),myeloid differentiation factor88 (MyD88) and related factors of Nuclear factor-kappa B (NF-κB) and Mitogen-activated protein kinases (MAPKs) pathways in the established model to induce SBD-1.And then four specific inhibitors of NF-κB and MAPKs pathways were chosen,which were PDTC (NF-κB signaling pathway inhibitor),SB202190 (P38 signaling pathway inhibitor),PD98059 (ERK1/2 signaling pathway inhibitor) and SP600125 (JNK signaling pathway inhibitor).Four kinds of inhibitors were used to treat cells by single or combined with each other and then further induced culture,while using RT-qPCR methods for detecting expression level of RECs SBD-1 mRNA after inhibitor treatment.The results showed that the cells stimulated by S.cerevisiae resulted in the transcription of NF-κB,P38,JNK,ERK1/2,TLR2 and MyD88 were significantly increased (P＜0.05).And PDTC,SB202190,SP600125 and PD98059 significantly inhibited (P＜0.01)the SBD-1 transcription of stimulated cells that previously treated with the inhibitor or inhibitors,and the SB202190 had the best inhibitory effect.The results suggest that the pathways of S.cerevisiae inducing SBD-1 transcription may be related to TLR2-MyD88-NF-κB/MAPKs,but the TLR2-MyD88-P38 of TLR2-MyD88-MAPKs may be as main signaling pathway.关键词
β-防御素-1/瘤胃上皮细胞/酿酒酵母菌/抑制剂/信号通路/实时荧光定量PCRKey words
β-defensin-1/ruminal epithelial cells/Saccharomyces cerevisiae/inhibitors/signaling pathways/real-time fluorescence quantitative PCR分类
农业科技引用本文复制引用
金鑫,张曼,田巧珍,刘骄,王云鹤,杨银凤..酿酒酵母菌诱导绵羊瘤胃上皮细胞SBD-1表达的信号通路初步研究[J].畜牧兽医学报,2017,48(3):561-569,9.基金项目
国家自然科学基金(31160491 （）
31560682) （）
