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

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (5): 1026-1032.doi: 10.11843/j.issn.0366-6964.2016.05.021

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

范燕茹^1，2，金鑫^1，2，田巧珍^1，2，张曼^1，2，刘骄^1，2，杨银凤^1，2*

（1.内蒙古农业大学兽医学院，呼和浩特 010018； 2.农业部动物疾病临床诊疗技术重点实验室，呼和浩特 010018）

收稿日期:2015-10-22 出版日期:2016-05-23 发布日期:2016-05-23
通讯作者: 杨银凤，教授，博士，主要从事动物解剖学与黏膜免疫研究，E-mail：julie1963@163.com
作者简介:范燕茹（1988-），女，内蒙古呼和浩特人，博士生，主要从事动物解剖学与黏膜免疫研究，E-mail: shining_fyr@126.com
基金资助:
国家自然科学基金（31160491）；内蒙古自治区人才开发基金项目

Preliminary Study on the Pathway of SBD-1 Expression in Sheep Rumen Epithelial Cells Induced by Lactobacillus plantarum

FAN Yan-ru^1.2，JIN Xin^1.2，TIAN Qiao-zhen^1.2，ZHANG Man^1.2，LIU Jiao^1.2，YANG Yin-feng^1.2*

（1.College of Veterinary，Inner Mongolia Agricultural University，Hohhot 010018，China；2.Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease，Ministry of Agriculture，Hohhot 010018，China）

Received:2015-10-22 Online:2016-05-23 Published:2016-05-23

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。

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.

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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参考文献

［1］SCHRÖDER J M，HARDER J.Human beta-defensin-2［J］.Int J Biochem Cell Biol，1999,31(6)：645-651.
［2］SØRENSEN O E,COWLAND J B,THEILGAARD-MÖNCH K,et al.Wound healing and expression of antimicrobial peptides/polypeptides in human keratinocytes，a consequence of common growth factors［J］. J Immunol，2003，170(11)：5583-5589.
［3］YANG Y F，WANG C Y，ZHAO Y F，et al.Reindeer beta-defensin-1:full-length cDNA cloning and tissue expression［J］. Vet Immunol Immunopathol,2009，131(1-2)：137-139.
［4］TANG Y Q，YUAN J，OSAPAY G，et al.A cyclic antimicrobial peptide produced in primate leukocytes by the ligation of two truncated α-defensins［J］. Science，1999，286(5439)：498-502.
［5］CHANG T L ，FRANÇ OIS F，MOSOIAN A，et al.CAF-mediated human immunodeficiency virus (HIV) type 1 transcriptional inhibition is distinct from α-defensin-1 HIV inhibition［J］.J Virol，2003，77(12)：6777-6784.
［6］HUTTNER K M，BREZINSKI-CALIGURI D J，MAHONEY M M，et al.Antimicrobial peptide expression is developmentally regulated in the ovine gastrointestinal tract［J］.J Nutr, 1998，128(2 Suppl)：297S-299S.
［7］HICKEY D K，FAHEY J V，WIRA C R.Mouse estrous cycle regulation of vaginal versus uterine cytokines，chemokines，α-/β-defensins and TLRs［J］.Innate Immun，2013,19(2)：121-131.
［8］DENG J，LI Y，ZHANG J，et al.Co-administration of Bacillus subtilis RJGP16 and Lactobacillus salivarius B1 strongly enhances the intestinal mucosal immunity of piglets［J］.Res Vet Sci，2013，94(1)：62-68.
［9］SCHLEE M，WEHKAMP J，ALTENHOEFER A，et al.Induction of human β-defensin 2 by the probiotic Escherichia coli Nissle 1917 is mediated through flagellin［J］. Infect Immun，2007，75(5)：2399-2407.
［10］PENHA FILHO R A，DÍAZ S J，FERNANDO F S，et al.Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic［J］. Vet Immunol Immunopathol，2015，167(1-2)：64-69.
［11］OZES O N，MAYO L D，GUSTIN J A，et al.NF-κB activation by tumour necrosis factor requires the Akt serine–threonine kinase［J］.Nature，1999，401(6748)：82-85.
［12］HERSHBERG R M.The epithelial cell cytoskeleton and intracellular trafficking.V.Polarized compartmentalization of antigen processing and Toll-like receptor signaling in intestinal epithelial cells［J］. Am J Physiol Gastrointest Liver Physiol，2002,283(4):G833-G839.
［13］GEWIRTZ A T.Intestinal epithelial toll-like receptors：to protect.And serve?［J］.Curr Pharm Des，2003,9(1):1-5.
［14］ABREU M T.Immunologic regulation of toll-like receptors in gut epithelium［J］.Curr Opin Gastroenterol，2003，19(6):559-564.
［15］RIZZO A，LOSACCO A，CARRATELLI C R.Lactobacillus crispatus modulates epithelial cell defense against Candida albicans through Toll-like receptors 2 and 4，interleukin 8 and human β-defensins 2 and 3［J］.Immunol Lett，2013，156(1-2)：102-109.
［16］温世勇.17-β雌二醇调控绵羊输卵管上皮细胞β-防御素1基因表达过程中NF-κB信号通路的研究［D］.呼和浩特：内蒙古农业大学,2014.
WEN S Y.Studys about NF-κβ signaling pathways in the course of the 17 β-estradiol regulating β-defensin gene 1 expression in the ovine oviduct epithelial cells［D］.Hohhot:Inner Mongolia Agricultural University，2014.(in Chinese)
［17］贾永杰.鼠李糖乳杆菌对鸡小肠上皮细胞β-防御素9基因表达的影响及其信号转导途径［D］.南昌：江西农业大学,2012.
JIA Y J.Effects of Lactobacillus rhamnosus on β-defensin 9 expression and its signal transduction pathway in chicken small intestinal epithelial cells［D］.Nanchang:Jiangxi Agricultural University，2012.(in Chinese)
［18］王佩,范燕茹,金鑫,等.枯草芽孢杆菌对绵羊瘤胃上皮细胞β-防御素-1表达的影响［J］.畜牧兽医学报,2015,46(5):760-767.
WANG P,FAN Y R,JIN X，et al.Effects of Bacillus subtilis on the expression of SBD-1 in cultured ruminal epithelial cells of sheep［J］.Acta Veterinaria et Zootechnica Sinica, 2015，46(5):760-767.(in Chinese)
［19］金鑫,张曼,范燕茹,等.酿酒酵母菌对绵羊瘤胃上皮细胞β-防御素-1(SBD-1)基因表达的影响［J］.中国农业科学,2015,48(19):3910-3918.
JIN X，ZHANG M，FAN Y R，et al.Effects of Saccharomyces cerevisiae on the expression of SBD-1 in cultured ruminal epithelial cells of sheep［J］ Scientia Agicultura Sinica，2015,48(19):3910-3918.(in Chinese)
［20］FUNDERBURG N T，SIEG S F.Diminished responsiveness to human β-defensin-3 and decreased TLR1 expression on monocytes and mDCs from HIV-1-infected patients［J］.J Leukoc Biol，2012，92(5)：1103-1109.
［21］HILL D R，KESSLER S P，RHO H K，et al.Specific-sized hyaluronan fragments promote expression of human β-defensin 2 in intestinal epithelium［J］.J Biol Chem，2012，287(36)：30610-30624.
［22］LI X Q，ZHU Y H，ZHANG H F，et al.Risks associated with high-dose Lactobacillus rhamnosus in an Escherichia coli model of piglet diarrhoea：intestinal microbiota and immune imbalances［J］. PLoS One，2012，7(7)：e40666.
［23］ZHU C，QIN H，CHENG T，et al.Staphylococcus aureus supernatant induces the release of mouse β-defensin-14 from osteoblasts via the p38 MAPK and NF-κB pathways［J］.Int J Mol Med，2013，31(6)：1484-1494.
［24］PAOLILLO R，ROMANO CARRATELLI C，SORRENTINO S，et al.Immunomodulatory effects of Lactobacillus plantarum on human colon cancer cells［J］.Int Immunopharmacol，2009，9(11)：1265-1271.

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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

Preliminary Study on the Pathway of SBD-1 Expression in Sheep Rumen Epithelial Cells Induced by Lactobacillus plantarum

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