【作者】 李兆龙；

【导师】 张有成；

【作者基本信息】 兰州大学 ， 普通外科学（专业学位）， 2021， 硕士

【摘要】 目的:旨在基于迷走神经回路(VN)研究胃肠动力障碍性疾病(DGIMD)可能的发病机制,并以厚朴三物汤(HPSWD)为整体,研究该方调节胃肠动力的作用靶点及分子机制,为厚朴三物汤的临床应用提供明确的分子生物学依据。方法:第一部分:将20只SD雄性大鼠随机分为对照组(10只)和模型组(10只),两组大鼠分别注射等体积的0.9%Nacl和左旋精氨酸(L-Arg)至腹腔,然后通过对每组大鼠的小肠推进率(SIPR)、血浆胃动素(MTL)和胃排空率(GER)进行测定来评估模型组胃肠动力障碍大鼠模型是否成功构建。第二部分:将40只SD雄性大鼠随机分为对照组(CG),模型组(MG),多潘立酮组(DG),厚朴三物汤组(HPSWD-G),每组各10只,模型组,多潘立酮组与厚朴三物汤组大鼠通过L-Arg腹腔注射法构建胃肠动力障碍大鼠模型,对照组大鼠注射等量0.9%Nacl溶液。造模成功并进行药物干预后,冰上取大鼠延髓,空肠,回肠,研究HPSWD对大鼠延髓迷走神经复合体(DVC)中γ-氨基丁酸(GABA)与其受体GABRA1,谷氨酸(Glu)与其受体NMDAR1;空肠,回肠神经递质乙酰胆碱(ACH)与其受体M2型乙酰胆碱受体(M2R),神经递质一氧化氮(NO),血管活性肠肽(VIP)以及它们的受体鸟苷酸环化酶(GC),血管活性肠肽受体1(VPAC1)表达的影响。结果:1.腹腔注射L-Arg 5天后,模型组大鼠的SIPR、血浆MTL、GER、体重较对照组明显下降。2.经药物干预7天后,与对照组相比,模型组大鼠延髓的兴奋性神经递质Glu及其受体NMDAR1表达下调,而抑制性神经递质GABA及其受体GABRA1表达上调;同时空肠与回肠的兴奋性神经递质ACH及其受体M2R表达下调,抑制性神经递质NO,VIP及其受体GC,VPAC1表达上调。3.与模型组相比,厚朴三物汤组大鼠延髓的兴奋性神经递质Glu及其受体NMDAR1表达上调,而抑制性神经递质GABA及其受体GABRA1蛋白表达下调;同时空肠和回肠的兴奋性神经递质ACH及其受体M2R表达上调,抑制性神经递质NO、VIP及其受体GC、VPAC1蛋白表达下调。4.多潘立酮组与厚朴三物汤组的VN相关神经递质及受体表达无差异。结论:1.大鼠的胃肠动力障碍模型可以通过左旋精氨酸的腹腔注射来构建。2.大鼠胃肠动力下降可能与延髓神经递质Glu与其受体NMDAR1表达下调,GABA与其受体GABRA1表达上调有关;同时也与空肠与回肠神经递质ACH及其受体M2R表达下调,神经递质NO,VIP及其受体GC,VPAC1表达上调有关。3.厚朴三物汤对胃肠动力的促进作用可能是通过上调延髓神经递质Glu与其受体NMDAR1表达水平,下调神经递质GABA与其受体GABRA1表达水平;同时上调空肠与回肠神经递质ACH及其受体M2R表达水平,下调神经递质NO,VIP及其受体GC,VPAC1表达水平实现的。更多还原

【Abstract】 Objective: To study the possible pathogenesis of disease of gastrointestinal motility disorders(DGIMD)based on the vagal neurocircuitry(VN),and to use Houpu Sanwu Decoction(HPSWD)as a whole to study its targets and molecules mechanism regulating gastrointestinal motility and provide a clear molecular biological basis for the clinical application of HPSWD.Method: Part 1: 20 SD male rats were randomly divided into a control group(10 rats)and a model group(10 rats).The two groups of rats were injected intraperitoneally with equal volumes of L-Arginine(L-Arg)and 0.9 %Nacl,then by measuring the small intestinal propulsion rate(SIPR),plasma motilin(MTL)and gastric emptying rate(GER)of each group of rats to evaluate whether the gastrointestinal motility disorder rat model of the model group was successfully constructed.Part 2: Randomly group 40 SD male rats into control group(CG),model group(MG),domperidone group(DG),Houpu Sanwu Decoction group(HPSWD-G),10 rats in each group.Rats in the model group,domperidone group and Houpu Sanwu decoction group were intraperitoneally injected with L-Arg to prepare the gastrointestinal motility dysfunction model.The rats in the control group were injected with the same amount of 0.9% Nacl solution.After successful modeling and drug intervention,the rats’ medulla oblongata,jejunum,and ileum were taken on ice to study the effects of HPSWD on rat medullary vagus nerve complex(DVC)γ-aminobutyric acid(GABA)and its receptor GABRA1,glutamate(Glu)and its receptor NMDAR1;jejunum,ileum neurotransmitter acetylcholine(ACH)and its receptor M2 acetylcholine receptor(M2R),neuro dorsal vagal complex transmitter nitric oxide(NO),vasoactive intestinal peptide(VIP)and their receptors guanylate cyclase(GC),vasoactive intestinal peptide receptor 1(VPAC1).Results: 1.After 5 days of intraperitoneal injection of L-Arg,compared with the control group,the SIPR,plasma MTL,GE and body weight of the model group rats were significantly reduced.2.After 7 days of drug intervention,compared with the control group,the expression of the neurotransmitter Glu and its receptor NMDAR1 in the medulla oblongata of the model group was down-regulated,while the expression of the neurotransmitter GABA and its receptor GABRA1 was up-regulated;the expression of neurotransmitter ACH and its receptor M2 R in the rat jejunum and ileum was downregulated,and the expression of neurotransmitter NO,VIP and their receptors GC,VPAC1 was up-regulated.3.Compared with the model group,the medulla oblongata excitatory neurotransmitter Glu and its receptor NMDAR1 protein expression increased,and the inhibitory neurotransmitter GABA and its receptor GABRA1 protein expression decreased;jejunum and ileum excitatory neurotransmitter ACH and its receptor M2 R protein expression increased,and inhibitory neurotransmitters NO,VIP and its receptor GC,VPAC1 protein expression decreased.4.There was no difference in the expression of VN-related neurotransmitters and receptors between domperidone group and Houpu Sanwu Decoction group.Conclusion: 1.Intraperitoneal injection of L-Arginine can successfully construct a rat model of gastrointestinal motility disorder.2.The down-regulation of the medullary neurotransmitter Glu and its receptor NMDAR1,and the up-regulation of the neurotransmitter GABA and its receptor GABRA1 are related to gastrointestinal motility disorders in the model group;it is also related to the down-regulation of neurotransmitter ACH and its receptor M2 R in the jejunum and ileum,and the upregulation of neurotransmitter NO,VIP and its receptor GC and VPAC1.3.The promoting effect of Houpu Sanwu Decoction on gastrointestinal motility may be through up-regulating the expression levels of the medullary neurotransmitter Glu and its receptor NMDAR1,down-regulating the expression levels of the neurotransmitter GABA and its receptor GABRA1;simultaneously up-regulating the jejunal and ileal neurotransmitters ACH and Its receptor M2 R expression level,down-regulate the neurotransmitter NO,VIP and its receptor GC,VPAC1 expression level.更多还原