益生菌干预对运动大鼠胃肠激素与AQP4表达的影响
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摘要
目的研究益生菌干预下力竭性运动大鼠胃肠道功能的影响。方法 40只SD雄性大鼠随机分为安静对照组(NC),运动对照组(ME),安静给药组(NCY),运动给药组(MEY)。ME与MEY组进行6周力竭训练,速度为19.3 m/min,坡度为5o,每天训练1次,每周6 d。NCY与MEY组每日灌胃浓度为10~7CFU/mL益生菌溶液10.0 mL/(kg·d)。6周力竭训练结束后,检测大鼠血清中胃肠激素ghrelin、PYY、CCK、GLP-1、MTL和GAS指标含量,并采用RT-PCR和Western blot法检测大鼠胃和结肠组织中AQP4的表达。结果 ME组大鼠血清ghrelin和PYY含量均高于NC组(P<0.01),而ME组CCK、GLP-1、MTL和GAS含量均低于NC组(P<0.01);NCY组大鼠血清ghrelin、PYY均低于ME组(P<0.01),而CCK、GLP-1、MTL和GAS含量均高于ME组(P<0.01);MEY组大鼠血清ghrelin高于NC组(P<0.05)而低于ME组(P<0.01);MEY组大鼠血清PYY的含量低于ME组(P<0.01);MEY组大鼠血清CCK含量低于NC组(P<0.05)而高于ME组(P<0.05);MEY组大鼠血清GLP-1、MTL和GAS含量均高于ME组(P<0.01)。ME组大鼠胃肠组织AQP4 mRNA和蛋白表达均低于NC组(P<0.01或P<0.05);NCY和MEY组大鼠胃肠组织AQP4 mRNA和蛋白表达均高于ME组(P<0.01或P<0.05)。结论力竭性运动引起大鼠胃肠激素含量发生变化以及AQP4表达的上升与补充益生菌溶液密切相关,提示益生菌可能参与了胃肠激素的分泌和水代谢的过程。
Objective To study the effect of intervention of probiotics on gastrointestinal function and expression of AQP4 in rats during exhaustive exercise. Methods Forty male SD rats were randomly divided into quiet control group( NC),exercise control group( ME),quiet medication group( NCY) and exercise medication group( MEY). The exercise control group and the exercise medication group were subjected to exhaustive training for 6 weeks: the running speed was 19. 3 m/min,the slope was 5°,the training was once a day,6 d/week for 6 weeks. Probiotics in concentration of 107 CFU/mL was gavaged 10. 0 mL/( kg/d) in the quiet medication group and exercise medication group. After 6 weeks of exhaustive training,the activities of serum gastrointestinal hormones ghrelin,PYY,CCK,GLP-1,MTL and GAS were measured.. The expression of AQP4 was detected by RT-PCR and western blot. Results The contents of serum ghrelin and PYY were significantly higher in the ME group than those in the NC group( P < 0. 01). The contents of intestinal CCK,GLP-1,MTL and GAS in the ME group were significantly lower than those in the NC group( P < 0. 01). The serum ghrelin and PYY were lower in the NCY group than in the ME group( P < 0. 01),while the contents of intestinal CCK,GLP-1,MTL and GAS in the NCY group were higher than those in the ME group( P < 0. 01). The serum ghrelin in the MEY group was higher than that in the NC group( P < 0. 05) and lower than that in the ME group( P < 0. 01).The PYY in the MEY group was lower than that in the ME group( P < 0. 05). The serum CCK was lower in the MEY group than that in the NC group( P < 0. 05) and higher than that in the ME group( P < 0. 05). The serum GLP-1,MTL and GAS in the MEY group were higher than those in the ME group( P < 0. 01). The expression of AQP4 mRNA and protein in the gastrointestinal tissue of the ME group were lower than those in the NC group( P < 0. 01 or P < 0. 05). The expression of AQP4 mRNA and protein in the NCY and MEY groups were higher than those in the ME group( P < 0. 01 or P < 0. 05). Conclusions Exhaustive exercise causes changes in gastrointestinal hormone levels and increase of AQP4 expression in rats. Probiotics may be involved in the gastrointestinal hormone secretion and water metabolic process.
Objective To study the effect of intervention of probiotics on gastrointestinal function and expression of AQP4 in rats during exhaustive exercise. Methods Forty male SD rats were randomly divided into quiet control group( NC),exercise control group( ME),quiet medication group( NCY) and exercise medication group( MEY). The exercise control group and the exercise medication group were subjected to exhaustive training for 6 weeks: the running speed was 19. 3 m/min,the slope was 5°,the training was once a day,6 d/week for 6 weeks. Probiotics in concentration of 107 CFU/mL was gavaged 10. 0 mL/( kg/d) in the quiet medication group and exercise medication group. After 6 weeks of exhaustive training,the activities of serum gastrointestinal hormones ghrelin,PYY,CCK,GLP-1,MTL and GAS were measured.. The expression of AQP4 was detected by RT-PCR and western blot. Results The contents of serum ghrelin and PYY were significantly higher in the ME group than those in the NC group( P < 0. 01). The contents of intestinal CCK,GLP-1,MTL and GAS in the ME group were significantly lower than those in the NC group( P < 0. 01). The serum ghrelin and PYY were lower in the NCY group than in the ME group( P < 0. 01),while the contents of intestinal CCK,GLP-1,MTL and GAS in the NCY group were higher than those in the ME group( P < 0. 01). The serum ghrelin in the MEY group was higher than that in the NC group( P < 0. 05) and lower than that in the ME group( P < 0. 01).The PYY in the MEY group was lower than that in the ME group( P < 0. 05). The serum CCK was lower in the MEY group than that in the NC group( P < 0. 05) and higher than that in the ME group( P < 0. 05). The serum GLP-1,MTL and GAS in the MEY group were higher than those in the ME group( P < 0. 01). The expression of AQP4 mRNA and protein in the gastrointestinal tissue of the ME group were lower than those in the NC group( P < 0. 01 or P < 0. 05). The expression of AQP4 mRNA and protein in the NCY and MEY groups were higher than those in the ME group( P < 0. 01 or P < 0. 05). Conclusions Exhaustive exercise causes changes in gastrointestinal hormone levels and increase of AQP4 expression in rats. Probiotics may be involved in the gastrointestinal hormone secretion and water metabolic process.
引文
[1]杨建昌,王军,陈乐琴,等.过度训练对大鼠胃肠道组织形态及功能的影响[J].中国体育科技,2005,41(5):123-126.Yang JC,Wang J,Chen LQ,et al.Effect of overtraining on gastrointestinal form and function of rat[J].China Sport sci Technol,2005,41(5):123-126.
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[7]莫伟彬,邱晓玲,李国峰.罗汉果甜苷对训练大鼠骨骼肌PGC-lαmRNA及蛋白表达的影响[J].现代预防医学,2017,44(1):131-134.Mo WB,Qiu XL,Li GF.MOG on the expression of PGC-lαmRNA and protein of skeletal muscle of endurance training ra t[J].Modern Prev Med,2017,44(1):131-134.
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[10]Gareau MG,Sherman PM,Walker WA.Probiotics and the gut microbiota in intestinal health and disease[J].Nat Rev Gastroenterol Hepatol,2010,7(9):503-514.
[11]Schwartz M,Regueiro M.Prevention and treatment of postoperative Crohn’s disease recurrence:Anupdate for a new decade[J].Curr Gastroenterol Rep,2011,13(1):95-100.
[12]Barbara G,Zecchi L,Barbaro R,et al.Mucosal permeability and immune activation as potential therapeutic targets of probiotics in irritable bowel syndrome[J].J Clin Gastroenterol,2012,46:S52-55.
[13]Jensen H,Grimmer S,Naterstad K,et al.In vitro testing of commercial and potential probiotic lactic acid bacteria[J].Int J Food Microbiol,2012,153(1-2):216-222.
[14]王茹,刘冬梅,吴娜娜,等.高住低练对肥胖青少年内源性大麻素及相关食欲调节激素的影响[J].体育科学,2016,36(2):51-57.Wang R,Liu DM,Wu NN,et al.The influences of“Living High-Training Low”on endocannabinoids and gastrointestinal hormones changes in obese adolescents[J].China Sport Sci,2016,36(2):51-57.
[15]王江,周永学,谢勇波.半夏泻心汤拆方对胃溃疡大鼠胃组织GAS、MTL与血清PAF、ET、NO的影响及其寒热并用配伍的意义研究[J].中华中医药学刊,2016,34(11):2776-2779.Wang J,Zhou YX,Xie YB.Effect of decomposed Banxia Xiexin decoction on GAS and MTL in gastric tissue and PAF,ET and NO in serum of gastric ulcer rats and its significance of cold and heat compatibility[J].Chin Arch Trad Chin Med,2016,34(11):2776-2779.
[16]Wu R,Dong W,Qiang X,et al.Orexigenic hormone ghrelin ameliorates gut barrier dysfunction in sepsis in rats[J].Crit Care Med,2009,37(8):2421-2426.
[17]Wu R,Dong W,Zhou M,et al.Ghrelin improves tissue perfusion in severe sepsis via downregulation of endothelin-1[J].Cardiovas Res,2005,68(2):318-326.
[18]Ashby D,Bloom SR.Recent progress in PYY research-An update report for 8th NPY meeting[J].Peptides,2007,28(2):198-202.
[19]Sloth B,Holst JJ,Flint A,et al.Effects of PYY1-36 and PYY3-36 on appetite,energy intake,energy expenditure,glucose and fat metabolism in obese and lean subjects[J].Am JPhysiol Endocrinol Metab,2007,292(4):E1062.
[20]Rosenbaum DM,Rasmussen SGF,Kobilka BK.The structure and function of G-protein-coupled receptors[J].Nature,2009,459(7245):356-363.
[21]Moreno JL,Holloway T,González-Maeso J.Chapter Eight-GProtein-coupled receptor heterocomplexes in neuropsychiatric disorders[J].Progr Mol Biol Transl Sci,2013,117:187-205.
[22]Ritzel R,Orskov C,Holst JJ,et al.Pharmacokinetic,insulinotropic,and glucagonostatic properties of GLP-1[7-36amide]after subcutaneous injection in healthy volunteers.Doseresponse-relationships[J].Diabetologia,1995,38(6):720-725.
[23]牛亭惠,吴国泰,陈振鹤,等.幼龄厌食模型大鼠胃肠激素变化规律研究进展[J].甘肃中医学院学报,2017,34(1):82-85.Niu TH,Wu GT,Chen ZH,et al.Research progress of changes of gastrointestinal hormones in young Rats with anorexia[J].J.Gansu Univ Chin Med,2017,34(1):82-85.
[24]Dockray G,Dimaline R,Varro A.Gastrin:old hormone,new functions[J].Pflügers Archiv Eur J Physiol,2005,449(4):344-355.
[25]常保荣,王小梅.有氧运动对老年大鼠胃肠动力及血清中胃促生长素、瘦素水平的影响[J].中国应用生理学杂志,2016,32(4):331-333.Chang BR,Wang XM.The influence of aerobic exercise on growth hormone,leptin levels in aged rats on gastrointestinal motility and stomach serum[J].Chin J Appl Physiol,2016,32(4):331-333.
[26]Westerterp-Plantenga MS.Decreased Glucagon-like peptide 1release after weight loss in overweight/obese subjects[J].Obesity,2005,13(4):710-716.
[27]Doucet E,Cameron J.Appetite control after weight loss:what is the role of bloodborne peptides?[J].Appl Physiol Nutr Metab,2007,32(3):523-532.
[28]Gueugnon C,Mougin F,Nguyen NU,et al.Ghrelin and PYYlevels in adolescents with severe obesity:effects of weight loss induced by long-term exercise training and modified food habits[J].Eur J Appl Physiol,2012,112(5):1797-1805.
[29]蒋波,孙丽霞.肠道益生菌对窒息新生儿血浆胃动素、胃泌素及降钙素基因相关肽水平的相关性分析[J].中国食物与营养,2014,20(7):80-83.Jiang B,Sun LX.Correlation analysis on the probiotics and plasma motilin,gastrin and calcitonin gene-related peptide of asphyxia neonatorum[J].Food Nutr China,2014,20(7):80-83.
[30]Martins C,Morgan LM,Bloom SR,et al.Effects of exercise on gut peptides,energy intake and appetite[J].J Endocrinol,2007,193(2):251-258.
[31]郑必海,李素芝,闫春城,等.高原急性缺氧致急性胃肠道黏膜损伤及其机制探讨[J].华南国防医学杂志,2011,25(1):4-7.Zheng BH,Li SZ,Yan CC,et al.Acute injury of gastrointestinal mucosa caused by acute hypobaric hypoxia and its mechanism[J].Military Med J South China,2011,25(1):4-7.
[32]Mccoy ES,Haas BR,Sontheimer H.Water permeability through aquaporin-4 is regulated by protein kinase C and becomes ratelimiting for glioma invasion[J].Neuroscience,2010,168(4):971-981.
[33]Nagelhus EA,Ottersen OP.Physiological roles of aquaporin-4in brain[J].Physiol Rev,2013,93(4):1543-1562.
[34]Hardin JA,Wallace LE,Wong JF,et al.Aquaporin expression is downregulated in a murine model of colitis and in patients with ulcerative colitis,Crohn’s disease and infectious colitis[J].Cell Tissue Res,2004,318(2):313-323
[35]杨鹏,徐华,张华,等.慢性胃炎不同病理类型与水通道蛋白3、水通道蛋白4表达的相关性[J].实用医学杂志,2015,31(15):2471-2474.Yang P,Xu H,Zhang H,et al.The correlations between the different pathological types gastritis and the expressions of AQP3and AQP4[J].J Pract Med,2015,31(15):2471-2474.
[36]赵琴,房芸,潘凌云,等.基于致炎毒性,胃和十二指肠AQP3,AQP4表达的藤黄炮制减毒机制研究[J].中国中药杂志,2016,41(9):1627-1634.Zhao Q,Fang Y,Pan LY,et al.Attenuation mechanism of gamboges during processing based on inflammatory toxicity and AQP3,AQP4 protein and mRNA expressions in rat gastric and duodenal tissues[J].China J Chin Mat Med,2016,41(9):1627-1634.
[2]FAO,WHO.Report of a joint FAO/WHO expert consultation on evaluation of health and nutritional proper ties of probiotics in food including powder milk with live lactic acid bacteria[M].Cordoba:Argentina,2001.
[3]Del PM,Morelli L,Strozzi GP,et al.Probiotics:from research to consumer.[J].Dig Liver Dis,2006,38(Suppl 2):S248-S255.
[4]Huang WC,Chen YM,Kan NW,et al.Hypolipidemic effects and safety of Lactobacillus Reuteri 263 in a hamster model of hyperlipidemia[J].Nutrients,2015,7(5):3767-3782.
[5]Chen YM,Wei L,Chiu YS,et al.Lactobacillus plantarum TWK10 supplementation improves exercise performance and increases muscle mass in mice[J].Nutrients,2016,8(4):205.
[6]Bedford TG,Tipton CM,Wilson NC,et al.Maximum oxygen consumption of rats and its changes with various experimental procedures[J].J Appl Physiol,1979,47(6):1278-1283.
[7]莫伟彬,邱晓玲,李国峰.罗汉果甜苷对训练大鼠骨骼肌PGC-lαmRNA及蛋白表达的影响[J].现代预防医学,2017,44(1):131-134.Mo WB,Qiu XL,Li GF.MOG on the expression of PGC-lαmRNA and protein of skeletal muscle of endurance training ra t[J].Modern Prev Med,2017,44(1):131-134.
[8]Malaguarnera G,Leggio F,Vacante M,et al.Probiotics in the gastrointestinal diseases of the elderly[J].J Nutr Health Aging,2012,16(4):402-410.
[9]Chassard C,Dapoigny M,Scott KP,et al.Functional dysbiosis within the gut microbiota of patients with constipated-irritable bowel syndrome[J].Alim Pharmacol Ther,2012,35(7):828-838.
[10]Gareau MG,Sherman PM,Walker WA.Probiotics and the gut microbiota in intestinal health and disease[J].Nat Rev Gastroenterol Hepatol,2010,7(9):503-514.
[11]Schwartz M,Regueiro M.Prevention and treatment of postoperative Crohn’s disease recurrence:Anupdate for a new decade[J].Curr Gastroenterol Rep,2011,13(1):95-100.
[12]Barbara G,Zecchi L,Barbaro R,et al.Mucosal permeability and immune activation as potential therapeutic targets of probiotics in irritable bowel syndrome[J].J Clin Gastroenterol,2012,46:S52-55.
[13]Jensen H,Grimmer S,Naterstad K,et al.In vitro testing of commercial and potential probiotic lactic acid bacteria[J].Int J Food Microbiol,2012,153(1-2):216-222.
[14]王茹,刘冬梅,吴娜娜,等.高住低练对肥胖青少年内源性大麻素及相关食欲调节激素的影响[J].体育科学,2016,36(2):51-57.Wang R,Liu DM,Wu NN,et al.The influences of“Living High-Training Low”on endocannabinoids and gastrointestinal hormones changes in obese adolescents[J].China Sport Sci,2016,36(2):51-57.
[15]王江,周永学,谢勇波.半夏泻心汤拆方对胃溃疡大鼠胃组织GAS、MTL与血清PAF、ET、NO的影响及其寒热并用配伍的意义研究[J].中华中医药学刊,2016,34(11):2776-2779.Wang J,Zhou YX,Xie YB.Effect of decomposed Banxia Xiexin decoction on GAS and MTL in gastric tissue and PAF,ET and NO in serum of gastric ulcer rats and its significance of cold and heat compatibility[J].Chin Arch Trad Chin Med,2016,34(11):2776-2779.
[16]Wu R,Dong W,Qiang X,et al.Orexigenic hormone ghrelin ameliorates gut barrier dysfunction in sepsis in rats[J].Crit Care Med,2009,37(8):2421-2426.
[17]Wu R,Dong W,Zhou M,et al.Ghrelin improves tissue perfusion in severe sepsis via downregulation of endothelin-1[J].Cardiovas Res,2005,68(2):318-326.
[18]Ashby D,Bloom SR.Recent progress in PYY research-An update report for 8th NPY meeting[J].Peptides,2007,28(2):198-202.
[19]Sloth B,Holst JJ,Flint A,et al.Effects of PYY1-36 and PYY3-36 on appetite,energy intake,energy expenditure,glucose and fat metabolism in obese and lean subjects[J].Am JPhysiol Endocrinol Metab,2007,292(4):E1062.
[20]Rosenbaum DM,Rasmussen SGF,Kobilka BK.The structure and function of G-protein-coupled receptors[J].Nature,2009,459(7245):356-363.
[21]Moreno JL,Holloway T,González-Maeso J.Chapter Eight-GProtein-coupled receptor heterocomplexes in neuropsychiatric disorders[J].Progr Mol Biol Transl Sci,2013,117:187-205.
[22]Ritzel R,Orskov C,Holst JJ,et al.Pharmacokinetic,insulinotropic,and glucagonostatic properties of GLP-1[7-36amide]after subcutaneous injection in healthy volunteers.Doseresponse-relationships[J].Diabetologia,1995,38(6):720-725.
[23]牛亭惠,吴国泰,陈振鹤,等.幼龄厌食模型大鼠胃肠激素变化规律研究进展[J].甘肃中医学院学报,2017,34(1):82-85.Niu TH,Wu GT,Chen ZH,et al.Research progress of changes of gastrointestinal hormones in young Rats with anorexia[J].J.Gansu Univ Chin Med,2017,34(1):82-85.
[24]Dockray G,Dimaline R,Varro A.Gastrin:old hormone,new functions[J].Pflügers Archiv Eur J Physiol,2005,449(4):344-355.
[25]常保荣,王小梅.有氧运动对老年大鼠胃肠动力及血清中胃促生长素、瘦素水平的影响[J].中国应用生理学杂志,2016,32(4):331-333.Chang BR,Wang XM.The influence of aerobic exercise on growth hormone,leptin levels in aged rats on gastrointestinal motility and stomach serum[J].Chin J Appl Physiol,2016,32(4):331-333.
[26]Westerterp-Plantenga MS.Decreased Glucagon-like peptide 1release after weight loss in overweight/obese subjects[J].Obesity,2005,13(4):710-716.
[27]Doucet E,Cameron J.Appetite control after weight loss:what is the role of bloodborne peptides?[J].Appl Physiol Nutr Metab,2007,32(3):523-532.
[28]Gueugnon C,Mougin F,Nguyen NU,et al.Ghrelin and PYYlevels in adolescents with severe obesity:effects of weight loss induced by long-term exercise training and modified food habits[J].Eur J Appl Physiol,2012,112(5):1797-1805.
[29]蒋波,孙丽霞.肠道益生菌对窒息新生儿血浆胃动素、胃泌素及降钙素基因相关肽水平的相关性分析[J].中国食物与营养,2014,20(7):80-83.Jiang B,Sun LX.Correlation analysis on the probiotics and plasma motilin,gastrin and calcitonin gene-related peptide of asphyxia neonatorum[J].Food Nutr China,2014,20(7):80-83.
[30]Martins C,Morgan LM,Bloom SR,et al.Effects of exercise on gut peptides,energy intake and appetite[J].J Endocrinol,2007,193(2):251-258.
[31]郑必海,李素芝,闫春城,等.高原急性缺氧致急性胃肠道黏膜损伤及其机制探讨[J].华南国防医学杂志,2011,25(1):4-7.Zheng BH,Li SZ,Yan CC,et al.Acute injury of gastrointestinal mucosa caused by acute hypobaric hypoxia and its mechanism[J].Military Med J South China,2011,25(1):4-7.
[32]Mccoy ES,Haas BR,Sontheimer H.Water permeability through aquaporin-4 is regulated by protein kinase C and becomes ratelimiting for glioma invasion[J].Neuroscience,2010,168(4):971-981.
[33]Nagelhus EA,Ottersen OP.Physiological roles of aquaporin-4in brain[J].Physiol Rev,2013,93(4):1543-1562.
[34]Hardin JA,Wallace LE,Wong JF,et al.Aquaporin expression is downregulated in a murine model of colitis and in patients with ulcerative colitis,Crohn’s disease and infectious colitis[J].Cell Tissue Res,2004,318(2):313-323
[35]杨鹏,徐华,张华,等.慢性胃炎不同病理类型与水通道蛋白3、水通道蛋白4表达的相关性[J].实用医学杂志,2015,31(15):2471-2474.Yang P,Xu H,Zhang H,et al.The correlations between the different pathological types gastritis and the expressions of AQP3and AQP4[J].J Pract Med,2015,31(15):2471-2474.
[36]赵琴,房芸,潘凌云,等.基于致炎毒性,胃和十二指肠AQP3,AQP4表达的藤黄炮制减毒机制研究[J].中国中药杂志,2016,41(9):1627-1634.Zhao Q,Fang Y,Pan LY,et al.Attenuation mechanism of gamboges during processing based on inflammatory toxicity and AQP3,AQP4 protein and mRNA expressions in rat gastric and duodenal tissues[J].China J Chin Mat Med,2016,41(9):1627-1634.