L-精氨酸对大鼠胃肠激素分泌及食欲的影响
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摘要
本试验旨在研究L-精氨酸(L-Arg)对大鼠胃肠道钙敏感受体(calcium-sensing receptor,CaSR)基因表达、胃肠激素分泌及采食量的影响,试验分为两个部分。试验I,选择32只SD雄性大鼠,随机分为4组,即对照组(灌胃0 mmol·kg~(-1) L-Arg·HCl),低剂量组(5 mmol·kg~(-1) L-Arg·HCl),中剂量组(10 mmol·kg~(-1) L-Arg·HCl)和高剂量组(20 mmol·kg~(-1) L-Arg·HCl),记录大鼠灌胃后不同时间点采食量,筛选显著影响大鼠采食的L-Arg最适剂量用于试验II。试验II,选择30只雄性大鼠随机分为两组,L-Arg·HCl高剂量组(20 mmol·kg~(-1))及对照组(0 mmol·kg~(-1)),灌胃大鼠一周(每天一次),记录大鼠体重变化及日采食量。试验结束后采集血液、胃、小肠及下丘脑组织,检测血液中胃肠激素水平,胃肠道CaSR及下丘脑食欲调节因子基因表达。结果显示:试验I,10和20 mmol·kg~(-1)的L-Arg·HCl分别在灌胃后的0～1 h和0～24 h显著降低大鼠采食量(P<0.05),其中20 mmol·kg~(-1)效果最显著;试验II,与对照组相比,20 mmol·kg~(-1) L-Arg·HCl显著降低了大鼠体增重及前两天的累加采食量(P<0.05),促进了胃肠道CaSR、厌食因子POMC基因表达和胆囊收缩素(CCK)分泌(P<0.05),胃泌素(gastrin)及葡萄糖依赖性促胰岛素多肽(GIP)的分泌虽有增加但差异不显著。相关分析结果显示,血清中CCK浓度与十二指肠、空肠CaSR及下丘脑POMC基因表达量呈显著正相关(P<0.05);GIP浓度与十二指肠CaSR及下丘脑POMC基因表达量呈显著正相关(P<0.05);GLP-1浓度与空肠CaSR及下丘脑POMC基因表达量有正相关的趋势(0.05 The objective of this study was to investigate the effects of L-arginine on feed intake, gastrointestinal hormone secretion and gene expression of calcium-sensing receptors(CaSR) in rats. Two experiments were conducted. In experiment I, thirty-two male Sprague-Dawley(SD) rats were randomly divided into 4 groups: a control group(oral gavage 0 mmol·kg~(-1) L-Arg·HCl), a low dose group(5 mmol·kg~(-1) L-Arg·HCl), a medium dose group(10 mmol·kg~(-1) L-Arg·HCl) and a high dose group(20 mmol·kg~(-1) L-Arg·HCl). The feed intake was recorded at different time points after gavage, and the indicated dose of L-Arg·HCl was chosen for the experiment II based on feed intake. In experiment II, thirty male rats were divided into two groups: a L-Arg·HCl treatment group(20 mmol·kg~(-1)) and a control group(0 mmol·kg~(-1)). The corresponding solution was gavaged once a day consecutively for one week. During this period, daily feed intake and changes in body weight were recorded. At the end of the experiment, all rats were euthanased. The venous blood, stomach and small intestine tissue, and hypothalamus were collected for the analysis of gastrointestinal hormones, and gene expression of the CaSR, neuropeptide Y and proopiomelanocortin(POMC) levels. In experiment I, compared with the control group, the concentrations of L-Arg·HCl at 10 and 20 mmol·kg~(-1) reduced food intake for 0-1 and 0-24 h, respectively, after gavage(P<0.05), with the greatest reduction in feed intake occurring at 20 mmol·kg~(-1). In experiment II, after being orally gavaged with 20 mmol·kg~(-1) L-Arg·HCl for 1 week, the body weight gain and cumulative feed intake in the first two days were decreased(P<0.05), while the secretion of cholecystokinin(CCK), expression of CaSR and POMC were up-regulated(P<0.05). However, the secretion of gastrin and glucose-dependent insulinotropic peptide(GIP) was not significantly affected, but did show an increasing trend. A correlation analysis suggested that the concentration of CCK in serum had a significant positive correlation with the gene expression level of CaSR in the duodenum, jejunum and with hypothalamic POMC(P<0.05), while the concentration of GIP in serum had a positive correlation with the expression level of CaSR in the duodenum and POMC in the hypothalamus(P<0.05). Meanwhile, the concentration of glucagon-like peptide-1 in serum displayed a trend towards positive correlation with the gene expression level of CaSR in jejunum and POMC in the hypothalamus(0.05
引文
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[17]Clemmensen C,Madsen A N,Smajilovic S,et al.L-Arginine improves multiple physiological parameters in mice exposed to diet-induced metabolic disturbances.Amino Acids,2012,43(3):1265-1275.
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[20]Wang C,Kang C,Xian Y,et al.Sensing of L-Arginine by gut-expressed calcium sensing receptor stimulates gut satiety hormones cholecystokinin and glucose-dependent insulinotropic peptide secretion in pig model.Journal of Food Science,2018,83(9):2394-2401.
[21]Alamshah A,McGavigan A K,Spreckley E,et al.L-arginine promotes gut hormone release and reduces food intake in rodents.Diabetes,Obesity and Metabolism,2016,18(5):508-518.
[22]Holst J J.The physiology of glucagon-like peptide 1.Physiological Reviews,2007,87(4):1409-1439.
[23]Sam A H,Troke R C,Tan T M,et al.The role of the gut/brain axis in modulating food intake.Neuropharmacology,2012,63(1):46-56.
[24]Sui X Y,Gao Q,Liu L,et al.Effects of dietary protein level on appetitive peptide gene expression of rabbits.Chinese Journal of Animal Nutrition,2015,27(9):2947-2954.隋啸一,高琴,刘磊,等.饲粮蛋白质水平对家兔食欲肽基因表达的影响.动物营养学报,2015,27(9):2947-2954.
[25]Fan W,Ellacott K L,Halatchev I G,et al.Cholecystokinin-mediated suppression of feeding involves the brainstem melanocortin system.Nature Neuroscience,2004,7(4):335-336.
[26]Chu S C,Chen P N,Ho Y J,et al.Both neuropeptide Y knockdown and Y1receptor inhibition modulate CART-mediated appetite control.Hormones&Behavior,2015,67:38-47.
[2]Liu J,Yu K,Zhu W.Amino acid sensing in the gut and its mediation in gut-brain signal transduction.Animal Nutrition,2016,2(2):69-73.
[3]Brennan S C,Davies T S,Schepelmann M,et al.Emerging roles of the extracellular calcium-sensing receptor in nutrient sensing:Control of taste modulation and intestinal hormone secretion.British Journal of Nutrition,2014,111(S1):16-22.
[4]Zhao X,Xian Y,Wang C,et al.Calcium sensing receptor mediated L-tryptophan-induced secretion of cholecystokinin and glucosedependent insulinotropic peptide in swine duodenum.Journal of Veterinary Science,2018,19(2):179-187.
[5]Xian Y,Zhao X,Wang C,et al.Phenylalanine and tryptophan stimulate gastrin and somatostatin secretion and H+-K+-ATPase activity in pigs through calcium-sensing receptor.General and Comparative Endocrinology,2018,267:1-8.
[6]Mace O J,Schindler M,Patel S.The regulation of K-and L-cell activity by GLUT2and the calcium-sensing receptor CasR in rat small intestine.The Journal of Physiology,2012,590(12):2917-2936.
[7]Prinz P,Stengel A.Control of food intake by gastrointestinal peptides:Mechanisms of action and possible modulation in the treatment of obesity.Journal of Neurogastroenterology and Motility,2017,23(2):180-196.
[8]Alamshah A,Spreckley E,Norton M,et al.l-phenylalanine modulates gut hormone release and glucose tolerance,and suppresses food intake through the calcium-sensing receptor in rodents.International Journal of Obesity,2017,41(11):1693-1701.
[9]Jordi J,Herzog B,Camargo S M,et al.Specific amino acids inhibit food intake via the area postrema or vagal afferents.The Journal of physiology,2013,591(22):5611-5621.
[10]Cheng S X,Okuda M,Hall A E,et al.Expression of calcium-sensing receptor in rat colonic epithelium:Evidence for modulation of fluid secretion.American Journal of Physiology-Gastrointestinal and Liver Physiology,2002,283(1):240-250.
[11]Yang K,Guan H,Arany E,et al.Neuropeptide Y is produced in visceral adipose tissue and promotes proliferation of adipocyte precursor cells via the Y1receptor.The FASEB Journal,2008,22(7):2452-2464.
[12]Chen C P,Kuhn P,Advis J P,et al.Chronic ethanol consumption impairs the circadian rhythm of pro-opiomelanocortin and period genes mRNA expression in the hypothalamus of the male rat.Journal of Neurochemistry,2004,88(6):1547-1554.
[13]Westerterpplantenga M S,Lejeune M P G M,Nijs I,et al.High protein intake sustains weight maintenance after body weight loss in humans.International Journal of Obesity,2004,28(1):57-64.
[14]Veldhorst M,Smeets A,Soenen S,et al.Protein-induced satiety:Effects and mechanisms of different proteins.Physiology&Behavior,2008,94(2):300-307.
[15]Greenwood H C.The role of specific amino acids in the regulation of food intake.London:Imperial College London,2011.
[16]Mentlein R,Gallwitz B,Schmidt W E.Dipeptidyl-peptidase IV hydrolyses gastric inhibitory polypeptide,glucagon-like peptide-1(7-36)amide,peptide histidine methionine and is responsible for their degradation in human serum.European Journal of Biochemistry,1993,214(3):829-835.
[17]Clemmensen C,Madsen A N,Smajilovic S,et al.L-Arginine improves multiple physiological parameters in mice exposed to diet-induced metabolic disturbances.Amino Acids,2012,43(3):1265-1275.
[18]Zhao X Y,Meng X L,Wu L,et al.Effects of dietary protein contents on CaSR expressions in small intestine and serum gastrointestinal hormone levels.Animal Husbandry&Veterinary Medicine,2016,48(12):30-35.赵秀英,孟祥龙,伍力,等.日粮不同蛋白水平对猪小肠CaSR基因表达及胃肠激素分泌的影响.畜牧与兽医,2016,48(12):30-35.
[19]Breer H,Eberle J,Frick C,et al.Gastrointestinal chemosensation:Chemosensory cells in the alimentary tract.Histochemistry Cell Biology,2012,138(1):13-24.
[20]Wang C,Kang C,Xian Y,et al.Sensing of L-Arginine by gut-expressed calcium sensing receptor stimulates gut satiety hormones cholecystokinin and glucose-dependent insulinotropic peptide secretion in pig model.Journal of Food Science,2018,83(9):2394-2401.
[21]Alamshah A,McGavigan A K,Spreckley E,et al.L-arginine promotes gut hormone release and reduces food intake in rodents.Diabetes,Obesity and Metabolism,2016,18(5):508-518.
[22]Holst J J.The physiology of glucagon-like peptide 1.Physiological Reviews,2007,87(4):1409-1439.
[23]Sam A H,Troke R C,Tan T M,et al.The role of the gut/brain axis in modulating food intake.Neuropharmacology,2012,63(1):46-56.
[24]Sui X Y,Gao Q,Liu L,et al.Effects of dietary protein level on appetitive peptide gene expression of rabbits.Chinese Journal of Animal Nutrition,2015,27(9):2947-2954.隋啸一,高琴,刘磊,等.饲粮蛋白质水平对家兔食欲肽基因表达的影响.动物营养学报,2015,27(9):2947-2954.
[25]Fan W,Ellacott K L,Halatchev I G,et al.Cholecystokinin-mediated suppression of feeding involves the brainstem melanocortin system.Nature Neuroscience,2004,7(4):335-336.
[26]Chu S C,Chen P N,Ho Y J,et al.Both neuropeptide Y knockdown and Y1receptor inhibition modulate CART-mediated appetite control.Hormones&Behavior,2015,67:38-47.