This trial was conducted to study the effect of Arg-Gly-Gln on jejunum enzyme activities, cell proliferation and apoptosis of weaned piglets in vitro. A single-factor design was adopted and six treatments were set, which were supplementation of Arg-Gly-Gln at 0, 0.28, 0.56, 1.11, 2.23 and 4.45 mmol/L in culture medium, respectively. There were eight replicates in each treatment and one jejunum tissue block in each replicate. The activity of lactate dehydrogenase（LDH） in culture medium, activities of glutaminase and diamine oxidase （DAO） in intestinal tissues, percentages of BrdU-positive cells and Caspase-3-positive cells were determined. The results showed as follows: 1) compared with that in culture medium without supplemental Arg-Gly-Gln, LDH activity in culture medium supplemented with Arg-Gly-Gln at different time point significantly decreased （P<0.05 or P<0.01）, excluding the culture medium supplemented with Arg-Gly-Gln at 0.56, 2.23 mmol/L at 6 h (P>0.05) and 1.11 mmol/L at 24 h (P>0.05); 2) compared with that in culture medium without supplemental Arg-Gly-Gln, glutaminase activity of jejunum tissue in culture medium supplemented with Arg-Gly-Gln at 1.11, 2.23 and 4.45 mmol/L increased by 27.69%, 46.15% and 83.08% (P<0.01), respectively, and DAO activity of jejunum tissue in culture medium supplemented with Arg-Gly-Gln at 2.23 and 4.45 mmol/L increased by 66.34% and 110.89% (P<0.01), respectively; 3) with the increasing of Arg-Gly-Gln levels in culture medium, percentage of BrdU-positive cells increased, but that of caspase-3-positive cells reduced. The results indicate that Arg-Gly-Gln can not only alleviate cell damage and reduce cell membrane permeability but also enhance the glutaminase and DAO activities, and promote the utilization of glutamine by intestinal cells. In addition, Arg-Gly-Gln can promote the cell proliferation of jejunum and inhibit apoptosis mediated by Caspase-3, resulting in the enhancement of barricade function of intestinal mucous membrane.［Chinese Journal of Animal Nutrition, 2011, 23（9）：1475 -1482］
JIANG Yi,JIA Gang,HUANG Lan,WU Caimei,WANG Kangning
. Effect of Different Arg-Gly-Gln Levels on Jejunum Enzyme Activities, Cell Proliferation and Apoptosis of Weaned Piglets in vitro[J]. Chinese Journal of Animal Nutrition, 2011
, 23(09)
: 1475
-1482
.
DOI: 10.3969/j.issn.1006-267x.2011.09.004
［1］LI Y S, LI J S, JIANG J W, et al. Glycyl-glutamine-supplemented long-term total parenteral nutrition selectively improves structure and function in heterotopic small-bowel autotransplantation in the pig[J]. Transplant International, 2003, 16(12):866-871.
[2]SCHEPPACH W, LOGES C, BARTRAM P, et al. Effect of free glutamine and alanyl-glutamine dipeptide on mucosal proliferation of the human ileum and colon[J]. Gastroenterology, 1994, 107(2):429-434.
[3]HAYNES T E, LI P, LI X, et al. L-glutamine or L-alanyl-L-glutamine prevents oxidant or endotoxin-induced death of neonatal entrecotes[J].Amino Acids, 2009, 37(1):131-142.
[4]BRANDSCH M, KNTTER I, LEIBACH F H. The intestinal H+/peptide symporter PEPT1: structure-affinity relationships[J]. European Journal of Pharmaceutical Sciences, 2004, 21(1):53-60.
[5]HUANG L, JIA G, WU C M, et al. The absorption and transportation of glutamine containing small peptides Arg-Gly-Gln and Gly-Gln in jejunum of weaned piglets[J]. Agricultural Science and Technology, 2010, 4(5):17-23.
[6]邱曙东,宋天保.组织化学与免疫组织化学[M].北京:科学技术出版社,2008.
[7]洪庆涛,宋岳涛,唐一鹏,等.细胞培养液乳酸脱氢酶漏出率的比色测定及其应用[J].细胞生物学杂志,2004,26(1):89-92.
[8]LEGRAND C, BOUR J M, JACOB C, et al. Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker[J]. Journal of Biotechnology, 1992, 25(3):231-243.
[9]KLIMBERG V S, SOUBA W W, SALLOUM R M, et al. Intestinal glutamine metabolism after massive small bowel resection[J]. American Journal of Surgery, 1990, 159(1):27-32.
[10]PINKUS L M, WINDMUELLER H G. Phosphate dependent glutaminase in the small intestine[J]. Archives of Biochemistry and Biophysics, 1977, 182(2):506-517.
[11]谢建新,顾岩,刘银坤,等.联合应用GH和Gln对短肠大鼠小肠黏膜上皮细胞谷氨酰胺酶和鸟氨酸脱羧酶活性的影响[J].解剖学杂志,2002,25(4):336-339.
[12]SATOH J, TSUJIKAWA T, FUJIYAMA Y, et al. Enteral alanyl-glutamine supplement promotes intestinal adaptation in rats[J]. International Journal of Molecular Medicine, 2003, 12(4):615-620.
[13]HAQUE S M, CHEN K, USUI N, et al. Alanyl-glutamine dipeptide-supplemented parenteral nutrition improves intestinal metabolism and prevents increased permeability in rats[J]. Annals of Surgery, 1996, 223(3):334-341.
[14]TAZUKE Y, WASA M, SHIMIZU Y, et al. Alanyl-glutamine-supplemented parenteral nutrition prevents intestinal ischemia-reperfusion injury in rats[J]. Journal of Parenteral and Enteral Nutrition, 2003, 27(2):110-115.
[15]BUFFONI F. Histaminase and related amine oxidases[J]. Pharmacological Reviews, 1966, 18(4):1163-1199.
[16]THOMPSON J S, VAUGHAN W P, FORST C F, et al. The effect of the route of nutrient delivery on gut structure and diamine oxidase levels[J]. Journal of Parenteral and Enteral Nutrition, 1987, 11(1):28-32.
[17]HESTERBERG R, SATTLER J, LORENZ W, et al. Histamine content, diamine oxidase activity and histamine methyltransferase activity in human tissues: fact or fictions[J]. Agents Actions, 1984, 14(3/4):325-334.
[18]FOGEL W A. Mucosal mono- and polyamine oxidase activities in digestive tract are distributed complementary to diamine oxidase[J]. Journal of Neural Transmission Supplementum, 1990, 32:345-349.
[19]LI J Y, LU Y, HU S, et al. Preventive effect of glutamine on intestinal barrier dysfunction induced by severe trauma[J]. World Journal of Gastroenterology, 2002, 8(1):168-171.
[20]VICARIO M, AMAT C, RIVERO M, et al. Dietary glutamine affects mucosal functions in rats with mild DSS-induced colitis[J]. The Journal of Nutrition, 2007, 137:1931-1937.
[21]WU G, MEIER S A, KNABLE D A. Dietary glutamine supplementation prevents jejunal atrophy in weaned pigs[J]. The Journal of Nutrition, 1996, 126:2578-2584.
[22]MCDOWELL E M, ZHANG X M, DESNATI A M. Localization of healing tracheal wounds using bromodeoxyuridine immunohistochemistry[J]. Stain Technology, 1990, 65:25-29.
[23]FAN M Z, STOLL B, JIANG R, et al. Enterocyte digestive enzyme activity along the crypt-villus and longitudinal axes in the neonatal pig small intestine[J]. Journal of Animal Science, 2001, 79:371-381.
[24]吴秀清，王虹，孙梅,等.幼年大鼠内毒素血症时小肠上皮细胞凋亡及Caspase-3的表达[J].中国当代儿科杂志，2005，7（2）：167-170.
[25]ERBIL Y, OZTEZCAN S, GIRI?瘙塁 M, et al. The effect of glutamine on radiation-induced organ damage[J]. Life Science, 2005, 78(4):376-382.
[26]SCHEPPACH W, DUSEL G, KUHN T, et al. Effect of L-glutamine and n-butyrate on the restitution of rat colonic mucosa after acid induced injury[J]. Gut, 1996, 38(6):875-885.
[27]BRASSE-LAGNEL C G, LAVOINNE A M, HUSSON A S. Amino acid regulation of mammalian gene expression in the intestine[J]. Biochimie, 2010, 92(7):729-735.
