黄酒通过肠肌间神经丛抑制大鼠离体小肠的收缩
|
摘要
目的:观察不同剂量的黄酒对大鼠离体小肠平滑肌收缩的影响并探究其可能的机制。方法:用低、中、高剂量的黄酒,乙醇,去多酚黄酒分别刺激大鼠离体小肠,RM6240多道生理信号采集系统记录大鼠离体小肠各节段加药前1 min和加药后第2分钟的收缩曲线,用肠道全层铺片免疫组织化学和Western blot技术检测大鼠肠肌间神经丛乙酰胆碱转移酶(choline acetyltransferase,ChAT)和一氧化氮合成酶(nitric oxide synthase,NOS)的表达。结果:与黄酒处理前相比较,中、高剂量(黄酒体积分数分别为3.5%、10.0%)的黄酒能明显抑制大鼠离体小肠的收缩幅度,抑制率分别为58%和100%;而中剂量的乙醇对离体小肠的收缩无明显的影响,高剂量的乙醇能显著抑制离体小肠的收缩幅度(抑制率为47.5%)。与处理前比较,经中、高剂量去多酚黄酒处理后,离体小肠运动幅度明显降低(P<0.05),抑制率分别为48%和67%。黄酒组肠肌间神经丛ChAT蛋白的表达量显著减少,而NOS蛋白的表达量显著增加。结论:黄酒对离体小肠收缩有明显的抑制作用,其作用与黄酒中的乙醇和多酚成分有关。黄酒可能通过抑制肌间神经丛中兴奋性神经递质乙酰胆碱的释放,促进抑制性神经递质一氧化氮的释放,而导致离体小肠的收缩受到抑制。黄酒能通过影响肌间神经丛内神经元的活性,抑制小肠运动,减缓胃肠机能亢进,为黄酒的保健功能研究及推广提供了理论依据。
Objective: The aim is to explore the effect and mechanism of Chinese rice wine(CRW) on the contraction activity of the isolated small intestine of rats. Methods: The isolated small intestine was treated separately with low, medium, high concentrations of CRW, alcohol and CRW without polyphenols. A RM6240 multi-channel physiological signal acquisition system was used to record intestinal motility. The protein expression of choline acetyltransferase(ChAT) and nitric oxide synthase(NOS) in intestinal myenteric plexus was measured by immunohistochemistry and Western blot. Results:Compared with the control group, medium and high-dose CRW(3.5% and 10.0%, V/V) obviously inhibited the amplitude of intestinal contraction with percentage inhibition of 58% and 100%, respectively. Medium-dose ethanol had no significant impact on intestinal contraction, but high-dose ethanol significantly inhibited the amplitude of intestinal contraction with a percentage inhibition of 47.5%. Medium and high-dose CRW without polyphenols significantly inhibited the contractile amplitude of the isolated small intestine by 48% and 67%, respectively, but the efficacy was inferior to that of CRW.Compared with the control group, the expression of ChAT in intestinal myenteric plexus in the CRW group significantly decreased, while the expression of NOS significantly increased. Conclusion: The effect of CRW on contraction of isolated small intestine is related to its ingredient ethanol and polyphenols, by inhibiting the release of acetylcholine(ACh) as an excitatory neurotransmitter and enhancing the release of NO as an inhibitory neurotransmitter from intestinal myenteric plexus. CRW can affect the activity of neurons in the myenteric plexus, inhibit the movement of small intestine, and slow down the hyperactivity of gastrointestinal function, which will provide new clues to the health-benefiting effects of CRW.
Objective: The aim is to explore the effect and mechanism of Chinese rice wine(CRW) on the contraction activity of the isolated small intestine of rats. Methods: The isolated small intestine was treated separately with low, medium, high concentrations of CRW, alcohol and CRW without polyphenols. A RM6240 multi-channel physiological signal acquisition system was used to record intestinal motility. The protein expression of choline acetyltransferase(ChAT) and nitric oxide synthase(NOS) in intestinal myenteric plexus was measured by immunohistochemistry and Western blot. Results:Compared with the control group, medium and high-dose CRW(3.5% and 10.0%, V/V) obviously inhibited the amplitude of intestinal contraction with percentage inhibition of 58% and 100%, respectively. Medium-dose ethanol had no significant impact on intestinal contraction, but high-dose ethanol significantly inhibited the amplitude of intestinal contraction with a percentage inhibition of 47.5%. Medium and high-dose CRW without polyphenols significantly inhibited the contractile amplitude of the isolated small intestine by 48% and 67%, respectively, but the efficacy was inferior to that of CRW.Compared with the control group, the expression of ChAT in intestinal myenteric plexus in the CRW group significantly decreased, while the expression of NOS significantly increased. Conclusion: The effect of CRW on contraction of isolated small intestine is related to its ingredient ethanol and polyphenols, by inhibiting the release of acetylcholine(ACh) as an excitatory neurotransmitter and enhancing the release of NO as an inhibitory neurotransmitter from intestinal myenteric plexus. CRW can affect the activity of neurons in the myenteric plexus, inhibit the movement of small intestine, and slow down the hyperactivity of gastrointestinal function, which will provide new clues to the health-benefiting effects of CRW.
引文
[1]倪莉,吕旭聪,黄志清,等.黄酒的生理功效及其生理活性物质研究进展[J].中国食品学报,2012,12(3):1-7.DOI:10.3969/j.issn.1009-7848.2012.03.001.
[2]王家林,张颖,于秦峰.黄酒中生物活性成分的探讨[J].酿酒科技,2011(7):47-50.
[3]谢广发.黄酒的功能性成分与保健功能[J].酿酒,2008,35(5):14-16.DOI:10.3969/j.issn.1002-8110.2008.05.007.
[4]沈赤.绍兴黄酒多糖的分离提取、生物活性及其对肠道微生物的影响[D].无锡:江南大学,2014:43-53.
[5]王璟,秦雪,仝令印,等.黄酒对小鼠抗疲劳能力和衰老小鼠免疫器官的影响[J].食品科学,2016,37(21):224-228.DOI:10.7506/spkx1002-6630-201621038.
[6]刘敬科,赵巍,付会期,等.小米黄酒酿造工艺的研究[J].粮食与饲料工业,2009(1 2):2 2-2 3;2 7.DOI:10.3 9 6 9/j.issn.1003-6202.2009.12.008.
[7]孙晖,田伟,张慧,等.葛根素对酒精中毒后家兔离体小肠平滑肌活动的影响[J].滨州医学院学报,2009,32(5):338-339;342.DOI:10.3969/j.issn.1001-9510.2009.05.006.
[8]张爱丽,刘姗姗,崔希云,等.石榴皮水提物对束缚浸水应激大鼠胃肠运动机能的影响[J].中国临床药理学杂志,2012,28(11):828-830.DOI:10.3969/j.issn.1001-6821.2012.11.009.
[9]赵梦杰,李春燕,李鹏飞,等.石榴皮水提物对大鼠离体结肠运动的影响及其机制的研究[J].生物医学工程研究,2010,29(2):109-112.DOI:10.3969/j.issn.1672-6278.2010.02.009.
[10]CAMILLERI M,FORD A C.Pharmacotherapy for irritable bowel syndrome[J].Journal of Clinical Medicine,2017,6(11):1-16.DOI:10.3390/jcm6110101.
[11]HERZLINGER M,CEREZO C.Functional abdominal pain and related syndromes[J].Child and Adolescent Psychiatric Clinics of North America,2018,27(1):15-26.DOI:10.1016/j.chc.2017.08.006.
[12]曾之晅,汪欢,杨新,等.肠易激综合征治疗研究进展[J].中华内科杂志,2017,56(2):140-143.DOI:10.3760/cma.j.issn.0578-1426.2017.02.014.
[13]DEFREES D N,BAILEY J.Irritable bowel syndrome:epidemiology,pathophysiology,diagnosis,and treatment[J].Primary Care,2017,44(4):655-671.DOI:10.1016/j.pop.2017.07.009.
[14]KU?AK-BEJDA A,BEJDA G,WASZKIEWICZ N.Antidepressants for irritable bowel syndrome:a systematic review[J].Pharmacological Reports,2017,69(6):1366-1379.DOI:10.1016/j.pharep.2017.05.014.
[15]HOLZER P,SCHICHO R,HOLZER-PETSCHE U,et al.The gut as a neurological organ[J].Wiener Klinische Wochenschrift,2001,113(17/18):647-660.
[16]吕锦芳,陈燕.不同处理因素对离体兔小肠平滑肌运动的影响[J].安徽技术师范学院学报,2002,16(1):17-22;28.DOI:10.3969/j.issn.1673-8772.2002.01.007.
[17]李林.乙酰胆碱酯酶阳性神经在大鼠小肠壁丛内分布的研究:酶组织化学法[J].江汉大学学报(自然科学版),1998(3):1-2.
[18]姜迎娅,陈忠.动物肠道主要神经递质及其受体分布与功能的研究进展[J].家畜生态学报,2013,34(9):1-7.
[19]王婧,潘盛武,杨盛昌.银杏叶提取物对小鼠离体小肠平滑肌收缩特性的影响[J].四川动物,2009,28(3):382-385.DOI:10.3969/j.issn.1000-7083.2009.03.015.
[20]FURNESS J B.Types of neurons in the enteric nervous system[J].Journal of the Autonomic Nervous System,2000,81(1):87-96.
[21]STEELE P A,BROOKES S J,COSTA M.Immunohistochemical identification of cholinergic neurons in the myenteric plexus of guinea-pig small intestine[J].Neuroscience,1991,45(1):227-239.DOI:10.1016/0306-4522(91)90119-9.
[22]FURNESS J B,CALLAGHAN B P,RIVERA L R,et al.The enteric nervous system and gastrointestinal innervation:integrated local and central control[J].Oxygen Transport to Tissue,2014,817:39-71.DOI:10.1007/978-1-4939-0897-4_3.
[23]张朝佑.人体解剖学(上册)[M].北京:人民卫生出版社,2009:1372-1381.
[24]彭曦,冯晋斌,汪仕良.大鼠消化道肌间神经丛中一氧化氮合酶的分布[J].世界华人消化杂志,1998(3):250-252.
[25]FURNESS J B.Integrated neural and endocrine control of gastrointestinal function[M]//The enteric nervous system.Berlin:Springer International Publishing,2016:1-15.DOI:10.1007/978-3-319-27592-5_16.
[26]袁园,林中,李小玲,等.肠神经系统重塑的研究进展[J].山东医药,2011,51(49):115-116.
[27]赵东芹,艾洪滨.束缚-浸水应激对大鼠迷走背核胆碱能神经元活动的影响[J].山东大学学报(医学版),2011,49(3):13-17.
[28]严海密,高锦孝,胡佩琪,等.消化性溃疡患者血清、胃液一氧化氮含量[J].世界华人消化杂志,1999,7(7):627-628.
[2]王家林,张颖,于秦峰.黄酒中生物活性成分的探讨[J].酿酒科技,2011(7):47-50.
[3]谢广发.黄酒的功能性成分与保健功能[J].酿酒,2008,35(5):14-16.DOI:10.3969/j.issn.1002-8110.2008.05.007.
[4]沈赤.绍兴黄酒多糖的分离提取、生物活性及其对肠道微生物的影响[D].无锡:江南大学,2014:43-53.
[5]王璟,秦雪,仝令印,等.黄酒对小鼠抗疲劳能力和衰老小鼠免疫器官的影响[J].食品科学,2016,37(21):224-228.DOI:10.7506/spkx1002-6630-201621038.
[6]刘敬科,赵巍,付会期,等.小米黄酒酿造工艺的研究[J].粮食与饲料工业,2009(1 2):2 2-2 3;2 7.DOI:10.3 9 6 9/j.issn.1003-6202.2009.12.008.
[7]孙晖,田伟,张慧,等.葛根素对酒精中毒后家兔离体小肠平滑肌活动的影响[J].滨州医学院学报,2009,32(5):338-339;342.DOI:10.3969/j.issn.1001-9510.2009.05.006.
[8]张爱丽,刘姗姗,崔希云,等.石榴皮水提物对束缚浸水应激大鼠胃肠运动机能的影响[J].中国临床药理学杂志,2012,28(11):828-830.DOI:10.3969/j.issn.1001-6821.2012.11.009.
[9]赵梦杰,李春燕,李鹏飞,等.石榴皮水提物对大鼠离体结肠运动的影响及其机制的研究[J].生物医学工程研究,2010,29(2):109-112.DOI:10.3969/j.issn.1672-6278.2010.02.009.
[10]CAMILLERI M,FORD A C.Pharmacotherapy for irritable bowel syndrome[J].Journal of Clinical Medicine,2017,6(11):1-16.DOI:10.3390/jcm6110101.
[11]HERZLINGER M,CEREZO C.Functional abdominal pain and related syndromes[J].Child and Adolescent Psychiatric Clinics of North America,2018,27(1):15-26.DOI:10.1016/j.chc.2017.08.006.
[12]曾之晅,汪欢,杨新,等.肠易激综合征治疗研究进展[J].中华内科杂志,2017,56(2):140-143.DOI:10.3760/cma.j.issn.0578-1426.2017.02.014.
[13]DEFREES D N,BAILEY J.Irritable bowel syndrome:epidemiology,pathophysiology,diagnosis,and treatment[J].Primary Care,2017,44(4):655-671.DOI:10.1016/j.pop.2017.07.009.
[14]KU?AK-BEJDA A,BEJDA G,WASZKIEWICZ N.Antidepressants for irritable bowel syndrome:a systematic review[J].Pharmacological Reports,2017,69(6):1366-1379.DOI:10.1016/j.pharep.2017.05.014.
[15]HOLZER P,SCHICHO R,HOLZER-PETSCHE U,et al.The gut as a neurological organ[J].Wiener Klinische Wochenschrift,2001,113(17/18):647-660.
[16]吕锦芳,陈燕.不同处理因素对离体兔小肠平滑肌运动的影响[J].安徽技术师范学院学报,2002,16(1):17-22;28.DOI:10.3969/j.issn.1673-8772.2002.01.007.
[17]李林.乙酰胆碱酯酶阳性神经在大鼠小肠壁丛内分布的研究:酶组织化学法[J].江汉大学学报(自然科学版),1998(3):1-2.
[18]姜迎娅,陈忠.动物肠道主要神经递质及其受体分布与功能的研究进展[J].家畜生态学报,2013,34(9):1-7.
[19]王婧,潘盛武,杨盛昌.银杏叶提取物对小鼠离体小肠平滑肌收缩特性的影响[J].四川动物,2009,28(3):382-385.DOI:10.3969/j.issn.1000-7083.2009.03.015.
[20]FURNESS J B.Types of neurons in the enteric nervous system[J].Journal of the Autonomic Nervous System,2000,81(1):87-96.
[21]STEELE P A,BROOKES S J,COSTA M.Immunohistochemical identification of cholinergic neurons in the myenteric plexus of guinea-pig small intestine[J].Neuroscience,1991,45(1):227-239.DOI:10.1016/0306-4522(91)90119-9.
[22]FURNESS J B,CALLAGHAN B P,RIVERA L R,et al.The enteric nervous system and gastrointestinal innervation:integrated local and central control[J].Oxygen Transport to Tissue,2014,817:39-71.DOI:10.1007/978-1-4939-0897-4_3.
[23]张朝佑.人体解剖学(上册)[M].北京:人民卫生出版社,2009:1372-1381.
[24]彭曦,冯晋斌,汪仕良.大鼠消化道肌间神经丛中一氧化氮合酶的分布[J].世界华人消化杂志,1998(3):250-252.
[25]FURNESS J B.Integrated neural and endocrine control of gastrointestinal function[M]//The enteric nervous system.Berlin:Springer International Publishing,2016:1-15.DOI:10.1007/978-3-319-27592-5_16.
[26]袁园,林中,李小玲,等.肠神经系统重塑的研究进展[J].山东医药,2011,51(49):115-116.
[27]赵东芹,艾洪滨.束缚-浸水应激对大鼠迷走背核胆碱能神经元活动的影响[J].山东大学学报(医学版),2011,49(3):13-17.
[28]严海密,高锦孝,胡佩琪,等.消化性溃疡患者血清、胃液一氧化氮含量[J].世界华人消化杂志,1999,7(7):627-628.