This experiment was conducted to study the effects of glucagon-like peptide-2 (GLP-2) supplementation on genes expression of intestinal epithelial tight junction proteins in weaner piglets and explore the possible molecular mechanism of GLP-2 regulating intestinal mucosal barrier of weaner piglets in vitro. The mRNA relative expression levels of ZO-1, Occludin, Claudin-1, MEK1/2 and p90RSK genes were investigated by placing jejunal tissues in culture medium with different GLP-2 concentrations (0, 1?10-8 and 1?10-7 mol/L). After the dose of GLP-2 which could promote the gene expression of key tight junction protein was determined, the changes in the mRNA relative expression levels of ZO-1, Occludin and Claudin-1 genes were studied by adding the specific inhibitor U0126 into the culture medium to block the mitogen-activated protein kinase (MAPK) signaling pathway. The results showed as follows: compared with the control group, when the jejunum of weaner piglets was cultured in vitro for 72 h in the medium with GLP-2 at the concentrations of 1?10-8 and 1?10-7 mol/L, the mRNA relative expression levels of MEK1/2, p90RSK, ZO-1, Occludin and Claudin-1 genes were increased significantly (P<0.05); except ZO-1 gene, the mRNA relative expression levels of all the other genes were increased with the increasing of GLP-2 concentration (P<0.05). When the U0126 was added to the culture medium with 1?10-7 mol/L GLP-2, compared with the group only added GLP-2, the mRNA relative expression levels of ZO-1, Occludin and Claudin-1 genes were significantly reduced (P<0.05). The results indicate that GLP-2 can increase the mRNA expression of ZO-1, Occludin and Claudin-1 genes, and the MAPK pathway may be one of the important signaling pathways for GLP-2 to modulate the genes expression of the tight junction proteins in intestine of weaner piglets.
JIANG Yi
,
JIA Gang
,
HUI Mingdi
,
CHEN Xiaoling
,
LI Hua
,
WANG Kangning
. Effects of Glucagon-Like Peptide-2 Supplementation on Expression of Intestinal Epithelial Tight Junction Protein Related Genes in Weaner Piglets in Vitro[J]. Chinese Journal of Animal Nutrition, 2012
, 24(9)
: 1785
-1792
.
DOI: 10.3969/j.issn.1006-267x.2012.09.023
[1] TSUKITA S,FURUSE M,ITOH M. Multifunctional strands in tight junctions[J].Nature Reviews Molecular Cell Biology,2001,2(4):285-293.
[2] 刘彩萍,胡彩虹,徐勇.早期断奶对仔猪肠道通透性和肠上皮紧密连接蛋白Occludin mRNA表达的影响[J].动物营养学报,2008,20(4):442-446.
[3] PETERSON Y M,HARTMAN B,HOLST J J,et al.Introduction of enteral food increases plasma GLP-2 and decreases GLP-2 receptor mRNA abundance during pig development[J].The Journal of Nutrition,2003,133(6):1781-1786.
[4] BURRIN D G,PETERSON Y,STOLL B,et al.Glucagons-like peptide-2:a nutrient responsive gut growth factor[J].The Journal of Nutrition,2001,131(3):709-712.
[5] CAMERON H L,PERDUE M H.Stress impairs murine intestinal barrier function:improvement by glucagons-like peptide-2[J].The Journal of Pharmacology and Experimental Therapeutics,2005,314(1):214-220.
[6] 关莉莉,宫德正,田楠,等.胰高血糖素样肽-2对小鼠小肠缺血/再灌注损伤的保护作用[J].中国应用生理学杂志,2005,21(2):192-195.
[7] BENJAMIN M A,MCKAY D M,YANG P C,et al.Glucagons-like peptide-2 enhances intestinal epithelial barrier function of both Tran cellular and Para cellular pathways in the mouse[J].Gut,2000,47:112-119.
[8] 陈振勇,冯闲松,杨鹏,等.类高血糖素多肽-2对大鼠阻梗性黄疸小肠上皮紧密连接的调控[J].中国现代普通外科进展,2009,1(12):3-7.
[9] LORENZA G M,TAPIA R,CHAMORRO D.Crosstalk of tight junction components with signaling pathways[J].Biochimica et Biophysica Acta,2008,1778:729-756.
[10] 蒋荣川,贾刚,王康宁.胰高血糖素样肽-2对体外培养的断奶仔猪小肠黏膜上皮细胞形态、增殖及其酶活力的影响[J].动物营养学报,2008,20(6):699-705.
[11] PFAFFL M W.A new mathematical model for relative quantification in real time RT-PCR [J].Nucleic Acids Research,2001,29:2002-2007.
[12] SAKAKIBARA A,FURUSE M,SAITOU M,et al.Possible involvement of phosphorylation of occluding in tight junction formation[J].The Journal of Cell Biology,1997,137(6):1393-1401.
[13] MAZZON E,STURNIOLO G C,PUZZOLO D,et al.Effect of stress on the paracellular barrier in the rat ileum[J].Gut,2002,51:507-513.
[14] MORITA K,HURUSE M,FUJIMOTO K,et al.Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands[J].Proceedings of the National Academy of Sciences,1999,96:511-516.
[15] ZHONG Y,SAITOH T,MINASE T,et al.Monoclonal antibody 7H6 reacts with a novel tight junction associated protein distinct from ZO-1,cingulin and ZO-2[J].The Journal of Cell Biology,1993,120:477-483.
[16] YANG R K,TOMOYUKI H,LI J Y.Bile modulates intestinal epithelial barrier function via an extracellular signal related kinase 1/2 dependent mechanism[J].Intensive Care Medicine,2005,31(5):709-717.
[17] BRUEWER M,LUEGERING A,KUCHARZIK T,et al.Proinfiammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms[J].The Journal of Immunology,2003,171(61):64-72.
[18] ZHANG B K,GUO Y M.Supplemental zinc reduced intestinal permeability by enhancing occluding and zonula occludens protein-1 (ZO-1) expression in weaning piglets[J].British Journal of Nutrition,2009,102:687-693.
[19] BJERKNES M, CHENG H.Modulation of specific intestinal epithelial progenitors by enteric neurons [J]. Proceedings of the National Academy of Sciences of the United States of America,2001,98:12497-12502.
[20] GUAN X,KARPEN H E,STEPHENS J,et al.GLP-2 receptor localizes to enteric neurons and endocrine cells expressing vaso-active peptides and mediates increased blood fiow[J].Gastroenterology,2006,130:150-164.
[21] BURRIN D G,STOLL B,GUAN X F,et al.GLP-2 rapidly activates divergent intracellular signaling pathways involved in intestinal cell survival and proliferation in neonatal piglets[J].American Journal of Physiology,Endocrinology and Metabolism,2007,292:E281-E291.
[22] JASLEEN J,ASHLEY S W,SHIMODA N,et al.Glucagons-like peptide 2 stimulates intestinal epithelial proliferation in vitro[J].Digestive Diseases and Sciences,2002,47:1135-1140.
[23] YUSTA B,SOMWAR R,WANG F,et al.Identification of glucagons-like peptide-2 (GLP-2) activated signaling pathways in baby hamster kidney fibroblasts expressing the rat GLP-2 receptor[J].Journal of Biological Chemistry,1999,274(30):459-467.
[24] DUBE P E,ROWLAND K J,BRUBAKER P L.Glucagons-like peptide-2 activates β-catenin signaling in the mouse intestinal crypt:role of insulin-like growth factor-Ⅰ[J].Endocrinology,2008,149(1):291-301.
[25] FAVATA M F,HORIUCHI K Y,MANOS E J,et al.Identification of a novel inhibitor of mitogen activated protein kinase[J].Journal of Biological Chemistry,1998,273(18):623-632.
[26] 郭晓,潘峻.ERK阻断剂U0126对白血病细胞K562细胞周期的作用及机制[J].第三军医大学学报,2008(5):393-395.
