Molecular Nutrition

Effects of Alanyl-Glutamine Dipeptide on Localization and Expression of Tight Junction Protein Occludin in Small Intestinal Epithelial Cells from Piglets

  • DENG Chenxi ,
  • WANG Zirui ,
  • YOU Jinming ,
  • QU Mingren ,
  • YE Yaling ,
  • XIN Xiangrong ,
  • PAN Ke
Expand
  • College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China

Received date: 2013-10-09

  Online published: 2014-03-04

Abstract

This experiment was conducted to evaluate the effects of alanyl-glutamine dipeptide (Ala-Gln) on localization and expression of tight junction protein occludin in small intestinal epithelial cells from piglets using IPEC-1 cells isolated from jejunal epithelium as the model. The IPEC-1 cells of the same generation were planted into 6-six-well cell culture plates and were incubated with DMEM high-glucose medium containing 0 (control group), 0.25, 0.50, 1.00, 2.00, 4.00 mmol/L Ala-Gln, respectively. When the cell fusion had reached more than 70%, the immunofluorescence localization and Western Blot of occludin were detected. The results showed as follows: 1) the positive fluorescence staining clouds in cytoplasm of cells were observed in the control group, but that was not obvious among the junction of cells. The fluorescence signals among the junction of cells appeared to be strengthen gradually and the cell outline appeared clearer with the increase of Ala-Gln concentration. Contrarily, the fluorescence signals in cytoplasm of cells were weakened gradually. 2) Compared with the control group, the relative expression level of occludin in IPEC-1 cells in supplemental groups was significantly increased (P<0.01) and a trend from increase to decrease was observed with the increase of Ala-Gln concentration. The relative expression level of occludin in 2.00 mmol/L group was significantly higher than that in other supplemental groups (P<0.01). Therefore, Ala-Gln can up-regulate the expression of tight junction protein occludin in small intestinal epithelial cells from piglets, and promote the formation of intercellular tight junction structure, and then strengthen the barrier function of intestinal mucosa in piglets.

Cite this article

DENG Chenxi , WANG Zirui , YOU Jinming , QU Mingren , YE Yaling , XIN Xiangrong , PAN Ke . Effects of Alanyl-Glutamine Dipeptide on Localization and Expression of Tight Junction Protein Occludin in Small Intestinal Epithelial Cells from Piglets[J]. Chinese Journal of Animal Nutrition, 2014 , 26(3) : 694 -700 . DOI: 10.3969/j.issn.1006-267x.2014.03.019

References

[1] FURUSE M, SASAKI H, FUJIMOTO K, et al.A single gene product, clauin-1 or 2, reconstitutes tight junction strands and recuits occluding in fibroblasts[J].The Journal of Cell Biology, 1998, 143(2):391-401.  

[2] 刘海萍, 胡彩虹, 徐勇.早期断奶对仔猪肠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] 马永贤, 蒋朱明, 白满喜, 等.创伤后肠黏膜屏障改变分子生物学机制的研究[J].中华外科杂志, 1995, 33(6):371-373.

[4] MURAKAMI A E, SAKAMOTO M I, NATALIM R M, et al.Supplementation of glutamine and vitamin E on the morphometry of the intestinal mucosa in broiler chickens[J].Poultry Science, 2007, 86(3):488-495.  

[5] 袁雪波, 马黎, 陈克嶙, 等.丙氨酰谷氨酰胺二肽对哺乳仔猪生长性能、小肠形态学和血清生化指标的影响[J].动物营养学报, 2011, 23(1):94-101.

[6] 刘铁民, 吴衍忠.谷氨酰胺对过度运动大鼠肠黏膜屏障功能变化的干预机制[J].四川体育科学, 2007(4):55-58.

[7] TSUKITA S, FURUSE M.Occludin and claudins in tight-junction strands:leading or supporting players[J].Trends in Cell Biology, 1999, 9(7):268-273.  

[8] 高志光, 秦环龙.肠上皮细胞紧密连接的生物学功能及在肠屏障中的作用[J].肠外与肠内营养, 2005, l12(5):299-302.

[9] KUBOTA K, FURUSE M, SASAKI H, et al.Ca2+-independent cell-adhesion activity of claudins, a family of integralmembrane proteins localized at tight junctions[J].Current Biology, 1999, 9(18):1035-1038.  

[10] SAKAKIBARA A, FURUSE M, SATIOU M.Possible involvement of phosphorylation of occludin in tight junction formation[J].Journal of Cell Biology, 1997, 137(6):1393-1401.  

[11] BJARNASON I, MACPHERSON A, HOLLANDER D.Intestinal permeability:an overview[J].Gastroenterology, 1995, 108(5):1566-1581.  

[12] 蒋建文, 黎介寿, 李幼生, 等.甘氨酰谷氨酰胺二肽对猪自体移植小肠的营养作用[J].中华外科杂志, 1999, 37(11):677-679.

[13] SHIMIZU M, SON D O.Food-derived peptides and intestinal functions[J].Current Pharmaceutical Design, 2007, 13(19):885-895.

[14] WU X Q, SHU L H, SUN M, et al.Effect of glutamine on apoptosis of the small intestine in young rats with endotoxemia and its mechanism[J].Chinese Journal of Contemporary Pediatrics, 2006, 8(6):496-498.

[15] MARGARITIS V G, FILOS K S, MICHALAKI M A, et al.Effect of oral glutamine administration on bacterial tanslocation, endotoxemia, liver and ileal morphology, and apoptosis in rats with obstructive jaundice[J].World Journal of Surgery, 2005, 29(10):1329-1334.  

[16] DE-SOUZA D A, GREENE L J.Intestinal permeability and systemic infections in critically ill patients:effect of glutamine[J].Critical Care Medicine, 2005, 33(5):1125-1135.  

[17] ASSIMAKOPOULOS S F, NIKOLOPOULOU V N, SCOPA C D, et al.Beneficial effects of glutamine on intestinal barrier function in obstructive jaundice[J].World Journal of Surgery, 2005, 29(7):935-936.  

[18] BOELENS P G, HOUDIJK A P, HAARMAN H J, et al.Glutamine-enriched enteral nutrition decreases infectious complications in trauma patients[J].American Journal of Clinical Nutrition, 2002, 76(1):253-254.

[19] KARINCH A M, PAN M, LIN C M, et al.Glutamine metabolism in sepsis and infection[J].Journal of Nutrition, 2001, 131(9):2535S-2538S, 2550S-2551S.

[20] POTSIC B, HOLLIDAY N, LEWIS P, et al.Glutamine supplementation and deprivation:effect on artificially reared rat small intestinal morphology[J].Pediatric Research, 2002, 52(3):430-436.  

[21] 崔巍, 闻颖, 董亚珞, 等.谷氨酰胺对体外培养肠上皮细胞屏障通透性的影响[J].世界华人消化杂志, 2008, 16(33):3729-3733.

[22] 潘璠, 刘成霞.谷氨酰胺对体外肠上皮细胞缺血再灌注损伤后occludin蛋白表达的影响[J].中华临床医师杂志, 2012, 6(10):2624-2627.

[23] 张宇飞, 嵇武, 黎介寿.Occludin蛋白调控机制及临床意义的研究进展[J].肠外与肠内营养, 2010, 17(3):175-178.
Outlines

/