基于胃肠动力探讨健脾方剂对FD脾虚证病证结合大鼠模型的作用研究

发布时间：2018-01-17 00:08

  本文关键词：基于胃肠动力探讨健脾方剂对FD脾虚证病证结合大鼠模型的作用研究　出处：《中国中医科学院》2017年硕士论文　论文类型：学位论文

  更多相关文章： 功能性消化不良 香砂六君子 脾虚Ⅰ号方 胃肠激素 脾虚证 胃肠动力

【摘要】：研究背景功能性消化不良(functional dyspepsia,FD)系一类临床上常见和多发的胃肠功能紊乱性疾病,主要表现为餐后饱胀不适、早饱、上腹痛和上腹烧灼感等,根据罗马ⅣV将其分为餐后不适综合征(Postprandial Distress syndrome,PDS)和上腹痛综合征(EpigastricPainSyndrome,EPS),其中餐后不适综合征主要表现为早饱及餐后饱胀感,上腹痛综合征主要为位于上腹部的疼痛或烧灼感。其在全球发病率可达11.5%～29.2%[1],影响了全球1/4以上的人口。其病理生理机制并不十分明确,目前西医专家多认为与胃肠动力异常、内脏敏感性增高、胃酸分泌过多、幽门螺杆菌感染、脑-肠轴、自主神经系统和胃肠激素、社会心理学因素等相关,主要通过促进胃肠动力、降低内脏高敏感、治疗HP感染、抑制胃酸分泌、保护胃黏膜及精神心理治疗等,多采用单靶点对症治疗。中医认为功能性消化不良上腹烧灼感主要与“嘈杂”相对应,“上腹痛”与“胃痛”、“胃脘痛”对应,而餐后不适与早饱主要与“痞满”相应,基本病机为脾胃升降失司,功能失调,胃气壅滞,以脾虚气滞为主,治疗以健脾理气为主,中医药在FD的治疗方面有其独特的优势,本研究通过采用健脾方剂通过在体及离体方式干预FD脾虚证病证结合大鼠模型,研究其作用机制。研究目的1.通过对脾虚Ⅰ号方及香砂六君子汤进行在体药效及药理学实验,观察健脾方剂对FD脾虚证病证结合模型大鼠胃肠动力的作用,探讨其治疗FD的可能作用机制;2.通过离体状态下给予FD脾虚证病证结合模型大鼠不同剂量健脾方剂进行干预,探讨健脾方剂对大鼠不同部位胃组织胃动力的直接影响;3.观察离体状态下不同健脾组合物对不同部位胃组织胃动力的直接影响。研究内容实验一、FD脾虚证病证结合大鼠模型的建立与评价目的:建立与评价FD脾虚证病证结合大鼠模型材料和方法:将30只雄性SD大鼠(乳鼠)随机分为正常组、单纯碘乙酰胺灌胃组(疾病组)、碘乙酰胺灌胃叠加小平台站立组(病证结合组),出生7日购进,3天适应性饲养。出生10日SD雄性乳鼠,正常组每日以2%蔗糖溶液灌胃;模型组每日以0.1%碘乙酰胺溶液灌胃,每只0.2ml,连续灌胃6天;大鼠3周龄时,剔除母鼠,分笼,至大鼠6周龄后,继续给予正常鼠饲料喂养;碘乙酰胺叠加小平台站立组每日18:00-8:00进行小平台站立,连续14天。造模结束后分别观察一般情况、体重、进食量、饮水量、抓力、糖水偏好情况、血清乳酸、D-木糖、淀粉酶含量等。结果:(1)一般情况:病证结合模型组大鼠体重、进食量、饮水量与正常组及疾病模型组相比,明显偏低(P0.05)。(2)抓力测定:病证结合模型组大鼠抓力与正常组和疾病模型组相比显著降低(P0.05)。(3)糖水偏好情况:病证结合模型组大鼠在糖水消耗量方面有所降低,但与正常组及疾病模型组相比,差异无统计学意义(P0.05)。(4)血清学指标:与正常组及疾病模型组比较,病证结合模型组血清D-木糖、淀粉酶含量显著降低(P0.05),血清乳酸含量显著增加(P0.01)。实验二、健脾方剂对FD脾虚证病证结合大鼠模型在体胃肠动力作用机制的研究目的:观察脾虚Ⅰ号方及香砂六君子汤对FD脾虚证模型大鼠胃肠动力的作用,从胃排空、小肠推进实验及血清胃肠激素水平探讨其治疗FD的可能作用机制。材料与方法:采取碘乙酰胺叠加小平台站立法建立FD脾虚证病证结合大鼠模型,实验动物随机分为正常组、模型组、西药组(多潘立酮组)、脾虚Ⅰ号方高剂量组、中剂量组、低剂量组、香砂六君子高剂量组、中剂量组、低剂量组。造模结束后,按每10ml·kg-1体重大鼠给药1ml以脾虚Ⅰ号方或香砂六君子或多潘立酮或蒸馏水灌胃14d。在此过程中,定时观察并记录大鼠体重、进食量、饮水量、抓力、毛色、活动等一般情况。治疗完成后,给予大鼠灌胃营养性半固体糊,计算各组大鼠胃内残留率及小肠推进比。麻醉状态下腹主动脉取血,用ELISA或比色法检测血清中淀粉酶、D-木糖、乳酸、胃动素(MTL)、胃泌素(GAS)、促胃生长素(Ghrelin)、胆囊收缩素(CCK)、生长抑素(SS)、血管活性肠肽(VIP)含量。结果:1 一般情况治疗前,与正常组比较,模型组及其余各组大鼠体重、进食量、饮水量、抓力均显著降低(P0.05)。与模型组比较,除正常组外,其余各组间大鼠体重、进食量、饮水量、抓力均无明显差异(P9.05)。治疗后,与模型组比较,西药组,香砂低、中、高剂量组,脾虚中、高剂量组大鼠体重显著升高(P0.05)。香砂高剂量组,脾虚高剂量组大鼠饮水量显著升高(P0.05)。香砂高、中、低剂量组,脾虚高、中、低剂量组大鼠进食量显著升高(P0.05)。西药组,香砂高、中剂量组,脾虚高、低剂量组大鼠抓力显著升高(P0.05)。与西药组比较,香砂高剂量组,脾虚高剂量组大鼠饮水量显著升高(P0.05)。脾虚高、中剂量组大鼠进食量显著升高(p0.05)。各组大鼠体重、抓力与西药组比较差异均无统计学意义(P0.05)。2胃内残留率和小肠推进比(1)与正常组比较,模型组大鼠胃内残留率显著高于正常组(p0.01);与模型组比较,西药组、脾虚低剂量、香砂高剂量组大鼠胃内残留率显著降低(P0.05);与西药组比较,脾虚中、高剂量组,香砂低、中剂量组大鼠胃内残留率显著高于西药组(P0.05)。(2)与正常组比较,模型组大鼠小肠推进比显著低于正常组(P0.05);与模型组比较,西药组、脾虚低、中、高剂量组、香砂高剂量组小肠推进比显著剂量增加(P0.05);与西药组比较,脾虚高剂量组小肠推进比显著高于西药组(P0.01),香砂低剂量小肠推进比显著低于西药组(P0.05)。3血清学指标(1)与正常组比较,模型组血清D-木糖含量显著低于正常组(P0.05);与模型组比较,西药组、脾虚低剂量组血清D木糖含量均显著高于模型组(P0.01);与西药组比较,各中药组血清D-木糖含量差异均无统计学意义(P0.05)。(2)与正常组比较,模型组血清淀粉酶含量显著降低于(P0.05);与模型组比较,西药组、脾虚中、高剂量组,香砂低、高剂量组血清淀粉酶含量均显著高于模型组(P0.05);与西药组比较,各中药组血清淀粉酶含量差异均无统计学意义(P0.05)。(3)与正常组比较,模型组血清乳酸含量模型组显著升高(P0.05);与模型组比较,给药后西药组、脾虚中、高剂量,香砂低、中剂量血清乳酸含量均显著低于模型组(P0.05);与西药组比较,各中药组血清乳酸含量差异均无统计学意义(P0.05)。(4)与正常组比较,模型组血清胃动素含量显著低于正常组(p0.01);与模型组比较,西药组、脾虚低、中、高剂量组,香砂低、中、高剂量组血清胃动素含量均高于模型组(p0.01);与西药组比较,脾虚中、高剂量组,香砂低、中剂量组血清胃动素含量均高于西药组(P0.05)。(5)与正常组比较,模型组血清胃泌素含量显著低于正常组(P0.01);与模型组比较,西药组、脾虚低、中、高剂量组,香砂低、中、高剂量组血清胃泌素含量均高于模型组(P0.01);与西药组比较,脾虚高剂量组血清胃泌素含量显著高于西药组(P0.05)。(6)与正常组比较,模型组血清促胃生长素含量显著低于正常组(P0.05);与模型组比较,西药组、脾虚中、高剂量组,香砂高剂量组血清促胃生长素含量显著高于模型组(p0.05);与西药组比较,香砂高剂量组、脾虚高剂量组血清促胃生长素含量显著高于西药组(P0.05)。(7)与正常组比较,模型组血清胆囊收缩素含量高于正常组,但差异无统计学意义(P0.05);与模型组比较,香砂低剂量组血清胆囊收缩素含量显著低于模型组(p0.05);与西药组比较,香砂高剂量组血清胆囊收缩素含量显著高于西药组(P005)(8)与正常组比较,模型组血清血管活性肠肽含量显著高于正常组(P0.05);与模型组比较,香砂高剂量组显著高于模型组(P0.05);与西药组比较,各中药组血清血管活性肠肽含量差异均无统计学意义(P0.05)(9)与正常组比较,模型组血清生长抑素含量显著高于正常组(P0.05);与模型组比较,香砂中剂量组血清生长抑素含量显著降低(P0.05);与西药组比较,各中药组血清生长抑素含量差异均无统计学意义(P0.05)实验三、健脾方剂对FD脾虚证模型大鼠离体组织胃动力的作用研究目的:通过观察脾虚Ⅰ号方及香砂六君子汤干预功能性消化不良脾虚证模型大鼠离体胃组织,从胃体、胃窦收缩幅度和频率及胃底张力角度,试图明确脾虚Ⅰ号方及香砂六君子汤离体调节胃动力的作用机制。材料与方法:采取碘乙酰胺叠加小平台法建立功能性消化不良脾虚证大鼠模型,实验动物分为正常组、模型组,造模后取正常组与模型组大鼠胃体、胃底、胃窦不同组织,比较模型组与正常组各部位收缩的差异,离体给予不同浓度的脾虚Ⅰ号方及香砂六君子汤,观察药物对模型胃体、胃窦收缩幅度和频率及胃底张力的影响。结果:(1)在离体状态下,模型大鼠的胃底张力显著高于正常组,差异有统计学意义(P0.05),模型大鼠的胃窦、胃体收缩幅度和频率均显著低于正常组,差异有统计学意义(P0.05)。(2)给予不同浓度的脾虚Ⅰ号方后可见FD模型大鼠胃底张力随浓度增加而增高,差异有统计学意义(p0.05),而香砂六君子的胃底张力随给药浓度而降低,差异有统计学意义(P0.05)。(3)给脾虚Ⅰ号方后胃窦收缩幅度随给药浓度而明显增强,差异有统计学意义(P0.05),其收缩频率无明显变化(P0.05),给予香砂六君子低浓度时可显著提高胃窦收缩幅度及频率(P0.05),当浓度达到一定剂量时则抑制胃窦收缩频率和幅度(P0.05)。(4)给脾虚1号方后胃体收缩幅度随给药浓度而减小,差异有统计学意义(P0.05),其收缩频率则明显增加,差异有统计学意义(P0.05),给予香砂六君子不同浓度时胃体收缩幅度及频率均明显减小,差异有统计学意义(P0.05),当浓度达到一定剂量时则抑制胃窦收缩频率和幅度(P0.05)。实验四、不同健脾药物组合对正常大鼠离体胃动力的作用研究目的:通过观察不同健脾药对对离体大鼠胃窦收缩幅度及胃底张力的作用,探讨离体状态下不同健脾药对对胃动力的调节作用。材料与方法:将实验动物分为四组,取大鼠胃窦、胃底不同组织,离体给予不同浓度的健脾药、健脾理气药、健脾化湿药、健脾温中药的药物组合,观察药物对胃窦收缩幅度及胃底张力的作用差异。结果:(1)中药健脾组、健脾化湿组、健脾温中组可显著提高胃底张力,差异有统计学意义(P0.05),健脾理气组可显著降低胃底张力,差异有统计学意义(P0.05)。(2)中药健脾组,健脾化湿组可增加胃窦收缩幅度,差异有统计学意义(P0.05),健脾理气组可显著减小胃窦收缩幅度,差异有统计学意义(P0.05),健脾温中组对胃窦作用效果无差异(P0.05)。研究结论1.碘乙酰胺灌胃叠加改良小平台法站立制作出的功能性消化不良动物模型符合脾虚证特征,具有胃动力障碍表现。2.健脾方剂可以改善FD大鼠胃肠道吸收功能及乳酸堆积造成的肢倦乏力状态。3.健脾方剂方可直接促进模型大鼠胃排空及小肠推进或通过改善胃肠激素,调节胃肠运动,其中脾虚Ⅰ号方的小肠推进作用优于香砂六君子。4.在离体状态下,健脾方剂可直接作用于胃组织而改善胃动力,健脾方剂及健脾药对从不同角度发挥其对胃肠动力的双向调节作用,且呈现剂量依赖性。
[Abstract]:The research background of functional dyspepsia (functional dyspepsia FD) is a kind of clinical common and multiple gastrointestinal functional disorders, mainly for distention, postprandial satiety, upper abdominal pain and abdominal burning sensation, according to the Rome V IV will be divided into postprandial distress syndrome (Postprandial Distress syndrome, PDS) and epigastric pain syndrome (EpigastricPainSyndrome, EPS), the postprandial distress syndrome mainly for early satiety and postprandial fullness, epigastric pain syndrome is mainly located in the upper abdominal pain or burning sensation in the world. Its incidence rate is up to 11.5% ~ 29.2%[1], the impact of the global 1/4 of the population. Its pathophysiological mechanism is not very clear, the current Western experts believe that with abnormal gastrointestinal motility, visceral hypersensitivity, excessive secretion of gastric acid, Helicobacter pylori infection, brain gut axis, autonomic nervous system and gastrointestinal hormone, social psychology Other related factors, mainly by promoting gastrointestinal motility, reduce visceral sensitivity, the treatment of HP infection, inhibition of gastric acid secretion, protect the gastric mucosa and psychological treatment, the use of single target therapy. Chinese medicine functional dyspepsia epigastric burning sensation and "noisy" corresponding "pain" and "stomach", "stomachache" correspondence, and postprandial discomfort and early satiety and "fullness", the basic pathogenesis is spleen and stomach dysfunction, functional disorders, stomach stagnation, spleen deficiency qi stagnation, spleen qi in the treatment, traditional Chinese medicine has its unique advantage in the treatment of FD. This study by in vitro and in vivo intervention FD spleen deficiency disease combined with syndrome rat model by using Jianpi prescription, study its mechanism of action. Objective: 1. in vivo pharmacodynamics and pharmacology experiment of spleen Decoction No.1 and observe the curative effect of Six Gentlemen Decoction. Jianpi prescription combined with gastrointestinal motility in rats with spleen deficiency of FD, to explore the possible mechanism of treatment of FD; 2. in vitro under the condition of FD of spleen deficiency disease combined with syndrome model rats with different doses of Jianpi prescription intervention, to explore the effect of Jianpi Recipe on direct rats in different parts of gastric tissue of gastric motility the direct effect; body under the condition of different composition in different parts of the spleen and stomach tissue of gastric motility observed from 3.. The contents of Experiment 1, establishment and evaluation of rat models of spleen deficiency combined with FD Objective: to establish and evaluate FD combination of disease and syndrome of spleen deficiency rats model of materials and methods: 30 male SD rats (rats) were randomly divided into normal group, simple iodoacetamide gavage group (disease group), intragastric administration of iodoacetamide superimposed small platform group (standing group of combined disease and syndrome), born 7 days of purchase, 3 days of adaptive feeding. 10 day old male SD rats, normal group Daily with 2% sucrose solution by gavage; model group daily with 0.1% iodoacetamide solution by gavage, each 0.2ml, by gavage for 6 days; when rats were 3 weeks old, were removed, cage, to rats after 6 weeks of age, continue to give the normal rat diet; iodoacetamide superimposed small standing group per day platform 18:00-8:00 is a small platform to stand for 14 consecutive days. After the modeling, in general, were observed in body weight, food intake, water intake, grip strength, sucrose preference, serum lactic acid, D- xylose, amylase content. Results: (1) general situation: the combination of disease and syndrome model rats, weight, food intake. The amount of water compared with the normal model group and disease group, was significantly lower (P0.05). (2) determination of grip: Rats Model grip strength significantly decreased compared with the normal group and disease group model combining disease with syndrome (P0.05). (3) sucrose preference: the model group rats decreased in syrup the consumption of combined disease and syndrome, but The normal model group and disease group were compared, the difference was not statistically significant (P0.05). (4) serum index: compared with the normal group and disease model group, model group serum D- xylose combined disease, amylase content was significantly reduced (P0.05), serum lactic acid content increased significantly (P0.01). In experiment two, Jianpi Recipe on FD spleen deficiency rat model with the objective of research body gastrointestinal motility mechanism: To observe the effect of spleen deficiency 1 recipe and the curative effect of Six Gentlemen Decoction on gastrointestinal motility in FD rats with spleen deficiency syndrome, from the gastric emptying, intestinal propulsion test and serum gastrointestinal hormone levels to explore the possible mechanism of treatment of FD materials and methods. PingTai Railway Station: take iodoacetamide superimposed small legislation to establish FD spleen deficiency disease combined with syndrome rat model, the experimental animal were randomly divided into normal group, model group, western medicine group (domperidone group), spleen 1 Fang Gao dose group, middle dose group, low dose group, 棣欑爞鍏�鍚涘瓙楂樺墏閲忕唬�

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