胰高血糖素样肽-2对羔羊胃肠道重量、瘤胃发酵及小肠上皮发育相关基因表达的影响

doi:10.11843/j.issn.0366-6964.2018.03.011

摘要/Abstract

摘要：

旨在研究胰高血糖素样肽-2（GLP-2）对羔羊胃肠道重量、瘤胃发酵及小肠上皮发育相关基因表达的影响。试验选择10只体况良好、胎次一致、体重相近的新生羔羊，随机分为两组，分别为对照组（0.5%的牛血清白蛋白，溶于生理盐水）和GLP-2组（50 μg·kg^-1BW GLP-2，溶于0.5%的牛血清白蛋白中），每组5只。在羔羊28日龄，开始进行皮下注射，每隔12 h注射1次，持续14 d。羔羊42日龄时，第一次注射2 h后进行屠宰。采集瘤胃液测定pH及挥发性脂肪酸（Volatile fatty acid，VFA）浓度，采集小肠上皮用于基因定量分析。研究结果显示：1）与对照组羔羊相比，GLP-2组羔羊体重和胃肠道重量无显著变化（P > 0.05），但羔羊空肠重（P =0.083）和回肠重（P=0.060）有升高的趋势。2）GLP-2注射显著升高了羔羊血浆GLP-2浓度（P=0.025），对血浆葡萄糖、瘤胃pH和VFA浓度无显著影响（P > 0.05）。3）GLP-2注射显著升高了十二指肠上皮Cyclin B1（P=0.001）、Cyclin D1（P=0.044）、CDK1（P=0.028）及CDK6（P=0.016）的mRNA表达，空肠上皮Cyclin A（P=0.028）、Cyclin B1（P=0.016）、CDK1（P=0.013）、CDK4（P=0.023）及CDK6（P=0.028）的mRNA表达和回肠上皮Cyclin A（P=0.025）、Cyclin D1（P=0.001）、CDK2（P=0.013）、CDK4（P=0.020）的mRNA表达。4）GLP-2组羔羊十二指肠上皮GCG（P=0.036）和IGF-1R（P=0.031），空肠上皮GCG（P=0.049）、IGF-1（P=0.027）、IGF-1R（P=0.036）及GLP-2R（P=0.011），回肠上皮GCG（P=0.025）、IGF-1（P=0.029）、IGF-1R（P=0.029）和GLP-2R（P=0.032）的mRNA表达显著高于对照组。结果显示，皮下注射GLP-2对羔羊体重、胃肠道重量和瘤胃内环境都没有显著影响，但对空肠重和回肠重有增加的趋势，升高了血浆中GLP-2浓度，显著影响小肠上皮发育相关基因的表达，促进了羔羊小肠的发育。研究结果将为促进羔羊小肠发育营养策略的制定提供新思路。

Abstract:

The objective of this study was to evaluate the effect of glucagon-like peptide-2 (GLP-2) on gastrointestinal weight, rumen fermentation and expression of small intestinal epithelial development-related genes in lambs. In the present study, 10 lambs were randomly assigned into 2 groups:control group (0.5% bovine serum albumin(BSA) in saline, n=5) and GLP-2 group (50 μg·kg^-1 BW GLP-2 in 0.5% bovine serum albumin, n=5). At 28 days-old, lambs were subcutaneously injected with BSA or GLP-2 once every 12 hours in the following 14 days. At 42 days-old, all lambs were slaughtered at 2 h after the first injection. Rumen fluid was collected for the analysis of rumen pH and VFA concentration, and the small intestine epithelium was used for quantitative analysis. The results showed that:1) The body weight and gastrointestinal weight of lambs in GLP-2 group were not significantly affected(P>0.05). GLP-2 treatment had a tendency to increase the weight of jejunum(P=0.083) and ileum(P=0.060). 2) GLP-2 injection significantly increased plasma GLP-2 concentration(P=0.025), while there was no significant change in plasma glucose, rumen pH and VFA concentration (P > 0.05) compared with control group. 3) Compared with the control group, GLP-2 injection significantly increased the mRNA expression of Cyclin B1 (P=0.001), Cyclin D1 (P=0.044), CDK1 (P=0.028), CDK6 (P=0.016) in duodenum epithelium and Cyclin A (P=0.028), Cyclin B1 (P=0.016), CDK1 (P=0.013), CDK4 (P=0.023), CDK6 (P=0.028) in jejunum epithelium, and Cyclin A (P=0.025), Cyclin D1 (P=0.001), CDK2 (P=0.013), CDK4 (P=0.020) in ileum epithelium. 4) The mRNA expression of GCG (P=0.036), IGF-1R (P=0.031) in duodenum epithelium and the mRNA expression of GCG (P=0.049), IGF-1 (P=0.027), IGF-1R (P=0.036) and GLP-2R (P=0.011) in jejunum epithelium, and the mRNA expression of GCG (P=0.025), IGF-1 (P=0.029), IGF-1R (P=0.029) and GLP-2R (P=0.032) in ileum epithelium in GLP-2 group were significantly higher than that in the control group.These results suggest that subcutaneous injection of GLP-2 have no significant effect on body weight, gastrointestinal weight and rumen environment in lambs. However, GLP-2 have a tendency to increase jejunum weight and ileum weight, elevate plasma GLP-2 concentration, significantly affect the expression of small intestinal epithelial development-related genes, and promote the development of the small intestine in lambs. These findings will provide new insight into nutritional strategies for promoting the development of small intestine in lambs.

中图分类号:

S826.5

刘理想, 孙大明, 毛胜勇, 刘军花. 胰高血糖素样肽-2对羔羊胃肠道重量、瘤胃发酵及小肠上皮发育相关基因表达的影响[J]. 畜牧兽医学报, 2018, 49(3): 549-558.

LIU Li-xiang, SUN Da-ming, MAO Sheng-yong, LIU Jun-hua. Effects of Glucagon-like Peptide-2 on Gastrointestinal Weight, Rumen Fermentation and Expression of Small Intestinal Epithelial Development-Related Genes in Lambs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 549-558.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.03.011

https://www.xmsyxb.com/CN/Y2018/V49/I3/549

参考文献

[1] FENG Y, DEMEHRI F R, XIAO W, et al. Interdependency of EGF and GLP-2 signaling in attenuating mucosal atrophy in a mouse model of parenteral nutrition[J]. Cell Mol Gastroenterol Hepatol, 2017, 3(3):447-468.
[2] CASTRO J J, MORRISON S Y, HOSSEINNI A, et al. Secretion of glucagon-like peptide-2 responds to nutrient intake but not glucose provision in milk-fed calves[J]. J Dairy Sci, 2016, 99(7):5793-5807.
[3] DUBÉ P E, BRUBAKER P L. Frontiers in glucagon-like peptide-2:multiple actions, multiple mediators[J]. Am J Physiol Endocrinol Metab, 2007, 293(2):E460-E465.
[4] KVIDERA S K, HORST E A, SANZ FERNANDEZ M V, et al. Characterizing effects of feed restriction and glucagon-like peptide 2 administration on biomarkers of inflammation and intestinal morphology[J]. J Dairy Sci, 2017, 100(11):9402-9417.
[5] 方媛媛. 胰高血糖素样肽-2与脓毒症肠道的保护[J]. 海南医学, 2017, 28(3):456-458.
FANG Y Y. Gut protection of glucagon like peptide-2 for sepsis[J]. Hainan Medical Journal, 2017, 28(3):456-458. (in Chinese)
[6] 贾刚, 邓秋红, 蒋荣川, 等. 胰高血糖素样肽-2和二肽基肽酶Ⅳ抑制剂联合使用对断奶仔猪肠上皮细胞的影响[J]. 畜牧兽医学报, 2017, 48(1):99-107.
JIA G, DENG Q H, JIANG R C, et al. Effects of the combination of Glucagon-like peptide-2 and Dipeptidyl Peptidase-IV inhibitors on intestinal epithelial cells of weaned piglets in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(1):99-107.(in Chinese)
[7] GORKA P, KOEALSKI Z M, PIETRZAK P, et al. Effect of sodium butyrate supplementation in milk replacer and starter diet on rumen development in calves[J]. J Physiol Pharmacol, 2009, 60(S3):47-53.
[8] TAYLOR-EDWARDS C C, BURRIN D G, MATTHEWS J C, et al. Expression of mRNA for proglucagon and glucagon-like peptide-2(GLP-2) receptor in the ruminant gastrointestinal tract and the influence of energy intake[J]. Domest Anim Endocrinol, 2010, 39(3):181-193.
[9] TAYLOR-EDWARDS C C, BURRIN D G, HOLST J J, et al. Glucagon-like peptide-2(GLP-2) increases small intestinal blood flow and mucosal growth in ruminating calves[J]. J Dairy Sci, 2011, 94(2):888-898.
[10] CONNOR E E, KAHL S, ELSASSER T H, et al. Glucagon-like peptide 2 therapy reduces negative effects of diarrhea on calf gut[J]. J Dairy Sci, 2013, 96(3):1793-1802.
[11] MORAN A W, AL-RAMMAHI M, ZHANG C, et al. Sweet taste receptor expression in ruminant intestine and its activation by artificial sweeteners to regulate glucose absorption[J]. J Dairy Sci, 2014, 97(8):4955-4972.
[12] DRUCKER D J, YUSTA B. Physiology and pharmacology of the enteroendocrine hormone glucagon-like peptide-2[J]. Annu Rev Physiol, 2014, 76(76):561-583.
[13] NRC. Nutrient requirements of small ruminants:sheep, goats, cervids and new world camelids[M]. Washington, D. C:National Academy press, 2007.
[14] 秦为琳. 应用气相色谱测定瘤胃挥发性脂肪酸方法的研究改进[J]. 南京农学院学报, 1982, 5(4):110-116.
QIN W L. Determination of rumen volatile fatty acids by means of gas chromatography[J]. Journal of Nanjing Agricultural College, 1982, 5(4):110-116.(in Chinese)
[15] WANG A, GU Z, HEID B, et al. Identification and characterization of the bovine G protein-coupled receptor GPR41 and GPR43 genes[J]. J Dairy Sci, 2009, 92(6):2696-2705.
[16] TSAI C H, HILL M, ASA S L, et al. Intestinal growth-promoting properties of glucagon-like peptide-2 in mice[J]. Am J Physiol, 1997, 273(1):E77-E84.
[17] LEI Q C, BI J C, WANG X Y, et al. GLP-2 prevents intestinal mucosal atrophy and improves tissue antioxidant capacity in a mouse model of total parenteral nutrition[J]. Nutrients, 2016, 8(1):33.
[18] BURRIN D G, STOLL B, GUAN X F, et al. Glucagon-like peptide 2 dose-dependently activates intestinal cell survival and proliferation in neonatal piglets[J]. Endocrinology, 2005, 146(1):22-32.
[19] GHATEI M A, GOODLAD R A, TAHERI S, et al. Proglucagon-derived peptides in intestinal epithelial proliferation[J]. Dig Dis Sci, 2001, 46(6):1255-1263.
[20] YANO S, TAKEHARA K, TAZAWA H, et al. Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging[J]. Cell Cycle, 2017, 16(5):406-414.
[21] NARASIMHA A M, KAULICH M, SHAPIRO G S, et al. Cyclin D activates the Rb tumor suppressor by mono-phosphorylation[J]. Elife, 2014, 3(3):e02872.
[22] LEE H J, JEDRYCHOWSKI M P, VINAYAGAM A, et al. Proteomic and metabolomic characterization of a mammalian cellular transition from quiescence to proliferation[J]. Cell Rep, 2017, 20(3):721-736.
[23] KANAKKANTHARA A, JEGANATHAN K B, LIMZERWALA J F, et al. Cyclin A2 is an RNA binding protein that controls Mre11 mRNA translation[J]. Science, 2016, 353(6307):1549-1552.
[24] LINDQVIST A, VAN ZON W, KARLSSON R C, et al. Cyclin B1-Cdk1 activation continues after centrosome separation to control mitotic progression[J]. PLoS Biol, 2007, 5(5):e123.
[25] BARR A R, HELDT F S, ZHANG T L, et al. A dynamical framework for the all-or-none G1/S transition[J]. Cell Syst, 2016, 2(1):27-37.
[26] ØRSKOV C, HARTMANN B, POULSEN S S, et al. GLP-2 stimulates colonic growth via KGF, released by subepithelial myofibroblasts with GLP-2 receptors[J]. Regul Pept, 2005, 124(1-3):105-112.
[27] DUBE P E, FORSE C L, BAHRAMI J, et al. The essential role of insulin-like growth factor-1 in the intestinal tropic effects of glucagon-like peptide-2 in mice[J]. Gastroenterology, 2006, 131(2):589-605.
[28] LEEN J L, IZZO A, UPADHYAY C, et al. Mechanism of action of glucagon-like peptide-2 to increase IGF-I mRNA in intestinal subepithelial fibroblasts[J]. Endocrinology, 2011, 152(2):436-446.
[29] JASLEEN J, ASHLEY S W, SHIMODA N, et al. Glucagon-like peptide 2 stimulates intestinal epithelial proliferation in vitro[J]. Dig Dis Sci, 2002, 47(5):1135-1140.
[30] CONNOR E E, BALDWIN R L Ⅵ, CAPUCO A V, et al. Characterization of glucagon-like peptide 2 pathway member expression in bovine gastrointestinal tract[J]. J Dairy Sci, 2010, 93(11):5167-5178.
[31] NEW D C, WONG Y H. Molecular mechanisms mediating the G protein-coupled receptor regulation of cell cycle progression[J]. J Mol Signal, 2007, 2:2.
[32] 卢劲晔. 日粮能量水平对山羊瘤胃上皮生长的影响及机理研究[D]. 南京:南京农业大学, 2012.
LU J Y. Effects of dietary energy intake on growth of rumen epithelium and its underlying mechanism in goats[D]. Nanjing:Nanjing Agricultural University, 2012.(in Chinese)