基于代谢组学辛开苦降法治疗功能性消化不良作用机制的研究

发布时间：2018-04-05 19:23

  本文选题：功能性消化不良　切入点：代谢组学　出处：《北京中医药大学》2016年博士论文

【摘要】：研究背景功能性消化不良(functional dyspepsia, FD)属于临床常见功能性胃肠病,具有病程漫长、病情易反复等临床特点,约有15%-20%的FD患者相关临床症状难以缓解。然而,传统治疗方法,包括生活方式调整、抑酸药、促动力药、抗焦虑抑郁药及幽门螺旋杆菌的根除等,临床疗效有限。约达50%的患者有寻求其他疗法,如补充替代医学(complementary and alternative medicine, CAM)的帮助。中药(Chinese herbal medicine, CHM)治疗FD在缓解症状、改善患者生活质量等方面具有一定优势。魏玮教授基于20多年临证经验,遵循辛开苦降法,创立中药胃康宁,经临床试验证实具有良好疗效。同时基于动物实验、体外分子实验及蛋白质组学等研究,证实中药胃康宁具有促进胃肠运动、调节内脏高敏感、调节胃壁肌电活动与自主神经功能以及改善抗氧化能力、能量代谢及线粒体的功能等作用。系统评价(system review, SR)是循证医学的重要研究方法及最佳证据来源,而荟萃分析(meta analysis)作为系统评价的研究类型之一,为临床研究及实践提供高等级证据支持,已为临床医学各专业广泛使用。代谢组学(metabonomics)作为系统生物学领域内研究生物体在不同因素影响下整体功能状态变化的一门科学,其所特有的整体观、动态观与中医复方研究中的指导思想,如整体观念、系统生物学的思考模式、复杂性科学等观念相契合,为应用现代科学技术手段研究传统中医药物质基础及作用机制提供了新的技术,逐渐成为中药复方研究领域内的新热点。研究目的基于1HNMR代谢组学技术研究夹尾刺激FD模型大鼠血清及尿液中内源性代谢物的变化,初步探索辛开苦降法代表方胃康宁干预FD的作用机制。研究方法1.文献综述及系统评价通过国内外文献分析,综述FD中西医临床论治及代谢组学在中药复方领域内研究的最新进展。同时采用荟萃分析对中药复方治疗FD的临床疗效及安全性进行评价。具体方法：检索Web of Science, PubMed, EMBASE, Cochrane Library,以及中国知网、中国生物医学文献数据库、维普数据库中文科技期刊数据库、万方数据资源系统等数据库中口服中药复方干预FD的随机、双盲、安慰剂对照试验,对纳入文献采用Cochrane风险偏倚评估工具评价质量,并使用Review Manager V.5.2.7软件进行Meta分析。2.代谢组学研究30只Wistar雄性大鼠随机分为正常组10只、模型组20只,对模型组大鼠采用夹尾刺激法进行FD模型复制,连续7天。模型成功后,随机抽取模型组中半数大鼠构成胃康宁中药组,参照临床剂量给予药物干预：胃康宁中药组给予胃康宁中药溶液灌胃14天,正常组、模型组给予蒸馏水灌胃14天。分别在造模第0、4、7天及灌胃第4、7、11、14天经代谢笼留取大鼠尿液,在造模14天后经腹主动脉取血留取大鼠血清。采用VARIAN 600MHz核磁共振波谱仪分别对大鼠尿液、血清样品进行检测,MestReNova7.1.0软件对核磁图谱进行处理,SIMCA-P+12.0软件进行多变量统计分析。选用主成分分析、偏最小二乘判别分析以及正交偏最小二成判别分析等分析方法进行数据处理,同时联合使用变量权重重要性排序值、相关系数进行差异代谢物的筛选。研究结果1.文献综述及系统评价FD西医治疗以对症治疗为主,遵循个体化治疗原则,其中以生活方式调整、抑酸剂、促动力药为一线治疗,以抗焦虑抑郁药、CAM为二线治疗,对于幽门螺旋杆菌感染者建议行根除治疗；中医药物治疗则主要从经典方剂、自拟效方、辨证施治、中西药结合等方面论治；代谢组学在中药复方领域内的研究主要集中基于中医证型、西医病症、病证结合研究以及针对药效机制、配伍规律、毒性评价、煎煮方法等研究。共纳入14篇文献(1424例患者),进行合并处理后,总计10篇研究纳入荟萃分析。(1)疗效评价总体症状缓解率中药高于安慰剂对照(RR 1.45,95% CI[1.31-1.60];Z= 7.29,P0.00001)、中药对于中医证候的总体改善优于安慰剂(RR 1.36,95%CI[1.23-1.50];Z=6.17,P0.00001)、中药较安慰剂能更好改善FD患者生活质量(SMD 0.30, 95%CI[0.15-0.45];Z=3.80,P=0.0001);(2)安全性评价不良反应事件的发生率在中药治疗组与安慰剂对照组间无明显差异(RR 1.06,95%CI[0.66-1.70];Z=0.23,P=0.82)。2.代谢组学研究尿液代谢分析发现：在夹尾刺激FD大鼠模型复制过程中,模型组与正常组尿液代谢轮廓相较,上调的差异代谢物有亮氨酸、乳酸、丙氨酸、二甲胺、苯乙酰甘氨酸、马尿酸、尿囊素、胞啶、酪氨酸、咪唑及苯丙氨酸,下调的差异代谢物有乙酸、乙酰谷氨酸、琥珀酸、α-酮戊二酸、柠檬酸及三甲胺。在中药胃康宁干预FD的过程中,中药胃康宁组与模型组尿液代谢信息相较,上调的差异代谢物有乙酰谷氨酸、葡萄糖、肌酸、马尿酸、葫芦巴碱及甲酸,下调的生物标志物有α-羟基戊酸、亮氨酸、丙氨酸、己二酸、甘氨酸、顺乌头酸及苯丙氨酸。血清代谢分析发现：在夹尾刺激FD大鼠模型复制过程中,模型组与正常组尿液代谢轮廓相较,上调的差异代谢物有异亮氨酸、亮氨酸、脯氨酸及甲醇,下调的差异代谢物有葡萄糖。在中药胃康宁干预FD的过程中,中药胃康宁组与模型组尿液代谢信息相较,上调的差异代谢物有葡萄糖,下调的差异代谢物有脯氨酸、谷氨酸、β-羟丁酸、柠檬酸、蛋氨酸、甘油、1-甲基组氨酸、苯丙氨酸及甲酸。在应激FD模型复制过程中,内源性代谢信息的扰动主要影响的代谢通路涉及：苯丙氨酸、酪氨酸和色氨酸生物合成,柠檬酸循环,苯丙氨酸代谢,缬氨酸、亮氨酸和异亮氨酸生物合成,乙醛酸和二羧酸代谢以及酪氨酸代谢。在中药胃康宁干预FD过程中,内源性代谢信息网络变化涉及的代谢通路有：乙醛酸和二羧酸代谢,苯丙氨酸、酪氨酸和色氨酸生物合成,甘氨酸、丝氨酸和苏氨酸代谢,苯丙氨酸代谢,缬氨酸、亮氨酸和异亮氨酸生物合成以及甘油酯代谢。研究结论1.通过文献综述、系统评价,认为FD有效药物治疗需整合中医、西医及循证医学证据等,实现中西医结合优势互补以有效改善FD,同时代谢组学在中药复方研究领域内应用前景广泛。2.夹尾刺激FD模型大鼠的血清及尿液代谢信息与正常大鼠相较,主要发生能量代谢、应激相关代谢及肠道菌群等方面的代谢信息改变；经辛开苦降法代表方胃康宁干预后,夹尾刺激FD模型大鼠被扰动的代谢信息向正常状态发生转归,提示中药胃康宁对FD的作用机制可能是通过影响能量代谢、应激相关代谢及肠道菌群等多个代谢通路,调节代谢紊乱,从而起到对于FD的治疗作用。
[Abstract]:The research background of functional dyspepsia (functional dyspepsia FD) is a common clinical functional gastrointestinal disease, has a long course and clinical features of repeated relapses, about FD of patients with clinical symptoms of 15%-20% is difficult to solve. However, the traditional treatment methods, including lifestyle changes, acid suppression drugs, prokinetic drugs, anti anxiety antidepressant drugs and Helicobacter pylori eradication therapy, clinical curative effect is limited. Approximately 50% of the patients had to seek other therapies, such as complementary and alternative medicine (complementary and alternative medicine, CAM) with the help of traditional Chinese medicine. (Chinese herbal medicine, CHM) in the treatment of FD is to alleviate the symptoms, improve the quality of life of patients has certain advantages such as Wei Wei. At 20 years of clinical experience based on the following xinkaikujiang method, founded Corelle herbal medicine, proved by clinical experiments has good curative effect. At the same time, based on animal experiments, in vitro experiments and protein molecules Group research, Corelle has confirmed herbal medicine to promote gastrointestinal motility, regulate visceral hypersensitivity, regulating gastric myoelectrical activity and autonomic nerve function and improve the antioxidant capacity, energy metabolism and mitochondrial function. The evaluation system (system review SR) is an important research method of evidence-based medicine and the best source of evidence, and meta analysis (meta analysis) as one of the types of research evaluation system, provides a high level of evidence to support clinical research and practice, has been widely used for clinical medical professional. Metabonomics (metabonomics) as a science within the field of systems biology of organisms under the influence of different factors overall functional changes, the overall view of the the unique, dynamic view and guiding ideology of Chinese medicine research, such as the overall concept, systems biology thinking, complexity science ideas fit for The application of modern scientific and technical means of the traditional Chinese medicine material basis and mechanism provides a new technology, gradually become a new focus of research in traditional Chinese medicine in the field of 1HNMR. Changes of metabonomics technique stimulate tail FD model of endogenous metabolites in serum and urine in rats based on the purpose of the study, a preliminary exploration of xinkaikujiang mechanism on behalf of Corelle square stomach intervention of FD. 1. analysis methods of literature review and evaluation system at home and abroad through the literature review, the FD in western medicine clinical treatment and the new research progress of metabonomics in medicine field. The meta analysis of clinical efficacy and safety of traditional Chinese Medicine in the treatment of FD was evaluated at the same time. Specific methods: Web Science PubMed, of search, EMBASE, Cochrane, Library, and China HowNet, China biomedical literature database, VIP database Chinese according to the number of scientific journals Library, Wanfang Data Resource Database in the oral administration of traditional Chinese medicine intervention FD randomized, double-blind, placebo-controlled trial, the meta-analysis using Cochrane tool for assessing the risk of bias of quality evaluation, and the Meta analysis of the.2. metabolomics study of 30 male Wistar rats were randomly divided into normal group 10 using Review Manager V.5.2.7 software, model group 20, model group of rats by tail clamp stimulus method established FD model for 7 consecutive days. After the success of the model, half of the rats in the model group were randomly selected to form gastric Corelle Chinese medicine group, according to clinical dose of drug intervention: Corelle gastric medicine group were given gastric gavage for 14 Corelle herbal solution day, the normal group, model group were given distilled water for 14 days respectively in the model. The first 0,4,7 days and 4,7,11,14 days after intragastric administration of the metabolic cages to collect urine from rats at 14 days after modeling abdominal aortic blood specimens from rat serum by VA. RIAN 600MHz nuclear magnetic resonance spectroscopy of rat urine and serum samples were detected on the NMR spectra of the MestReNova7.1.0 software, SIMCA-P+12.0 software in multivariate statistical analysis. Using principal component analysis, partial least squares discriminant analysis and orthogonal partial least squares discriminant analysis method to carry on the data processing, and combined use of variable weights of importance the value of screening, correlation coefficient of different metabolites. 1. the results of literature review and evaluation system of FD medicine treatment for symptomatic treatment, follow the principle of individualized treatment, including lifestyle changes, antacids, prokinetic drugs as first-line treatment with anti anxiety depression drug CAM as second-line treatment for eradication therapy Helicobacter pylori infection; Chinese medicine treatment mainly from classical prescriptions, with side effects, traditional Chinese medicine and Western medicine combined with syndrome differentiation, etc. Surface treatment; metabonomics in compound Chinese medicine in the field of research mainly based on TCM, western medicine disease, and for evaluation of drug mechanism, combining disease with syndrome of regularity of compatibility, toxicity, study on Decoction methods. A total of 14 articles were included (1424 patients), by combining treatment, a total of 10 studies included in the meta-analysis. (1) the curative effect evaluation of the overall remission rate of Chinese medicine was higher than that of placebo (RR 1.45,95% CI[1.31-1.60] Z=; 7.29, P0.00001), traditional Chinese medicine for general syndromes of traditional Chinese medicine is better than placebo (RR 1.36,95%CI[1.23-1.50]; Z=6.17, P0.00001), traditional Chinese medicine can better improve the quality of life in patients with FD than placebo (SMD 0.30, 95%CI[0.15-0.45]; Z=3.80, P=0.0001); (2) adverse events occurred in safety evaluation of Chinese medicine treatment group and placebo groups had no obvious difference (RR 1.06,95%CI[0.66-1.70]; Z=0.23, P=0.82).2. metabonomics Study on metabolism of urine analysis found: in the stimulate tail FD rats in model group and normal group urinary metabolic profile in different metabolites have increased leucine, lactic acid, alanine, two methylamine, phenylacetylglycine, hippuric acid, allantoin, cytidine, tyrosine, phenylalanine and imidazole, metabolites that are down regulated. Acetic acid, acetyl Glutamic acid, succinic acid, ketoglutaric acid, citric acid and trimethylamine. In the process of Chinese medicine intervention in gastric Corelle FD, Corelle herbal medicine group and model group compared with the urine metabolic information, increase the differences of metabolites have acetyl glutamic acid, glucose, creatine, hippuric acid, formic acid and trigonelline, down regulated biological markers of alpha hydroxy acid, leucine, alanine, glycine, adipic acid, CIS aconitic acid and phenylalanine. The serum metabolic analysis found: in the stimulate tail FD rats in model group, and positive Often group urinary metabolic profile by upregulation of different metabolites have isoleucine, leucine, proline and methanol, lowered the difference. In the process of Chinese medicine glucose metabolites have stomach Corelle intervention FD, Corelle herbal medicine group and model group compared the difference of urine metabolic information, increased glucose metabolites, downregulation of different metabolites have proline. Glutamic acid, beta hydroxybutyric acid, citric acid, methionine, glycerol, 1- methyl histidine, phenylalanine and formic acid. The stress model of FD replication process, endogenous metabolic information disturbance mainly affect the metabolic pathways involved: phenylalanine, tyrosine and tryptophan biosynthesis, the citric acid cycle, phenylalanine metabolism, valine, leucine and isoleucine biosynthesis. Glyoxylic acid and two carboxylic acid metabolism and tyrosine metabolism. In the traditional Chinese medicine intervention of gastric Corelle FD process, involving endogenous metabolic changes in Information Network The metabolic pathway: glyoxylic acid and two carboxylic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, glycine, serine and threonine metabolism, phenylalanine metabolism, valine, leucine and isoleucine biosynthesis and glyceride metabolism. Conclusion: 1. through literature review, evaluation system, FD effective drug therapy should be integrated Chinese medicine, western medicine and evidence-based medical evidence, to realize the combination of complementary advantages to improve the FD of traditional Chinese medicine and Western medicine, and metabonomics widely used.2. tail clamp stimulation of FD model rats serum and urine metabolic information compared with normal rats in the study of Chinese herbal compound in the field, mainly in energy metabolism, the metabolic information related to stress metabolism and intestinal microflora the change of xinkaikujiang method; the representative prescription of Weikang Ninggan prognosis, metabolic information stimulate tail FD model rats by disturbance to the normal state change It suggests that the action mechanism of Corelle on FD is probably to regulate the metabolic disorder through metabolic pathways such as energy metabolism, stress related metabolism and intestinal flora, thereby playing a role in the treatment of FD.

【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R259
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本文编号：1716165