植物乳杆菌诱导绵羊瘤胃上皮细胞SBD-1表达的信号通路途径初探

doi:10.11843/j.issn.0366-6964.2016.05.021

摘要/Abstract

摘要：

本试验旨在探索乳酸杆菌诱导绵羊瘤胃上皮细胞SBD-1表达的可能途径。采用实时荧光定量PCR（RT-qPCR）的方法，对已建立的诱导SBD-1表达模型中Toll样受体2（Toll like receptor 2，TLR2）及其相关因子的基因表达变化进行检测；然后选用3种信号通路抑制剂，即NF-κB信号通路抑制剂PDTC、ERK 1/2信号通路抑制剂PD98059和JNK信号通路抑制剂SP600125，将细胞分为8组：细胞组:不作处理；阳性对照组:只添加植物乳杆菌P-8（L.plantarum P-8）诱导；PDTC组:只添加PDTC预处理细胞（PDTC）；PDTC+ L.plantarum P-8组: PDTC+L.plantarum P-8诱导；PD98059 组：PD98059；SP600125组: SP600125；PD98059+ L.plantarum P-8组: PD98059+ L.plantarum P-8；SP600125+ L.plantarum P-8组: SP600125+ L.plantarum P-8。采用RT-qPCR的方法检测各组SBD-1 mRNA表达水平。结果表明，绵羊瘤胃上皮细胞被L.plantarum P-8诱导后，TLR2及其转接蛋白MyD88、NF-κB和ERK1/2、JNK各基因的mRNA水平都较空白组细胞内的表达极显著增加（P＜0.01）；添加抑制剂后再诱导，发现抑制剂PD98059和SP600125均能极显著（P＜0.01）抑制细胞内SBD-1 mRNA的表达，而PDTC仅能显著抑制（P＜0.05）SBD-1的表达。结果表明，L.plantarum P-8可促进绵羊瘤胃上皮细胞TLR2、MyD88、NF-κB、JNK和ERK1/2基因的mRNA表达；添加NF-κB信号通路抑制剂PDTC、ERK 1/2信号通路抑制剂PD98059和JNK信号通路抑制剂SP600125，可抑制L.plantarum P-8对SBD-1 mRNA的诱导作用。综上表明，L.plantarum P-8有可能通过激活绵羊瘤胃上皮细胞内NF-κB、JNK、ERK1/2等信号通路促进SBD-1的表达。

关键词: β-防御素-1, 瘤胃上皮细胞, 乳酸杆菌, Toll样受体2, 实时荧光定量PCR

Abstract:

This study aims to explore the possible approach that lactobacilli induce the expression of SBD-1 in sheep rumen epithelium cells. Real-time fluorescence quantitative PCR（RT-qPCR）was conducted in this study to determine mRNA expressive variation of the toll like receptor 2 (TLR2) and its related factors in the established model to induce SBD-1. Three signaling pathway inhibitors were chosen which named NF-κB signaling pathway inhibitor PDTC, ERK 1/2 signaling pathway inhibitor PD98059 and JNK signaling pathway inhibitor SP600125, respectively. The tested cells were divided into 8 groups : cell group: without treatment; positive groups:L.plantarum P-8; PDTC groups : PDTC pretreatment cells only; PDTC+ L.plantarum P-8 group:PDTC+L.plantarum P-8;PD98059 group:PD98059;SP600125 group:SP600125;PD98059+ L.plantarum P-8 group:PD98059+L.plantarum P-8;SP600125+L.plantarum P-8 group: SP600125+ L.plantarum P-8.RT-qPCR method was used for detecting the expression levels of SBD-1 mRNA. The results indicated that the mRNA expression of TLR2, MyD88, NF-κB, ERK1/2 and JNK had significant increase compared with the cell groups (P<0.01) after the sheep rumen epithelial cells were induced by L.plantarum P-8, PD98059 and SP600125 could significantly inhibited the mRNA expression of SBD-1 in cells (P<0.01) with pretreatment that adding the inhibitors and were induced by L.plantarum P-8, however, the expression of SBD-1 was only inhibited significantly by the inhibitor PDTC (P<0.05). The results in this paper implied that L.plantarum P-8 could promote the mRNA expression of TLR2, MyD88, NF-κB, JNK and ERK1/2. To add NF-κB signaling pathway inhibitor PDTC, ERK 1/2 signaling pathway inhibitor PD98059 and JNK signaling pathway inhibitor SP600125 could inhibit the effect of L.plantarum P-8 on inducing SBD-1 mRNA. Thus, L.plantarum P-8 could improve the SBD-1 expression by activating the signaling pathways of NF-κB, JNK and ERK1/2.

Key words: β-defensin-1, rumen epithelium cells, lactobacillus, toll-like receptor 2, real-time quantitative PCR

中图分类号:

S826
S852

范燕茹，金鑫，田巧珍，张曼，刘骄，杨银凤. 植物乳杆菌诱导绵羊瘤胃上皮细胞SBD-1表达的信号通路途径初探[J]. 畜牧兽医学报, 2016, 47(5): 1026-1032.

FAN Yan-ru，JIN Xin，TIAN Qiao-zhen，ZHANG Man，LIU Jiao，YANG Yin-feng. Preliminary Study on the Pathway of SBD-1 Expression in Sheep Rumen Epithelial Cells Induced by Lactobacillus plantarum[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(5): 1026-1032.

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