基于脑肠轴探讨肝郁脾虚证功能性消化不良多肽组学及疏肝健脾法干预的研究

Functional dyspepsia（FD）with liver stagnation and spleen deficiency is the most common in clinical of Traditional Chinese Medicine（TCM）. Shugan Jianpi therapy is characterized by "reactive multi-components, multi-target and multi-channel effect ". Peptidomics featuring overall and dynamic core thinking is coincide with "unity"and "dialectics" theories of TCM，and it is a new idea to introduce peptidomics into study of syndromes of TCM to discuss the nature and mechanism of liver stagnation and spleen deficiency with FD and to observe the effct of Chinese herb intervention. Based on brain-gut axis，using two-dimensional gel electrophoresis and mass spectrometry techniques, the project will detect polypeptide expression spectrum of hypothalamus, dorsal root, stomach, intestine in liver stagnation and spleen deficiency syndrome with FD rats，screen differently expressed peptides and then make locating expression analysis of aimed peptides by immunofluorescence technique to reveal the nature and pathogenesis of liver stagnation and spleen deficiency with FD. Additionally，we will detect the proliferation, apoptosis and mark peptide expression of primary culture cells after treatment with the recipe medicated serum of Shugan Jianpi method to further clarify the therapeutic action of Shugan Jianpi method on liver stagnation and spleen deficiency syndrome with FD and the underlying mechanisms, which aims to provide a new warning, markers for diagnosis and new drug targets for prevention and treatment of functional dyspepsia.

功能性消化不良中医临床以肝郁脾虚证最为多见，使用疏肝健脾法治疗具有 "活性多组分、作用多靶点多途径"的特点。多肽组学的"整体性、动态性"核心思维与中医学"整体观、辨证观"理论体系相吻合，应用多肽组学来研究功能性消化不良中医证的本质及中药复方作用机制是一个新的思路。本项目以脑肠轴为切入点，通过双向凝胶电泳和生物质谱等技术对肝郁脾虚证功能性消化不良大鼠下丘脑、脊髓、胃和肠这条轴线多肽组表达谱进行分析；筛选差异性表达肽，采用免疫荧光染色技术对其进行定位表达分析，以揭示肝郁脾虚证功能性消化不良的发病机制；用疏肝健脾法方药含药血清处理原代培养细胞，检测细胞增殖活力、细胞凋亡和标志肽类的表达，进一步阐明疏肝健脾法治疗肝郁脾虚证功能性消化不良的作用机制。旨在为功能性消化不良的预防诊治提供新的预警、诊断标志物及新的药物作用靶点。

功能性消化不良（FD）是临床上常见的功能性胃肠紊乱疾病，属典型的身心疾病，心理-社会因素对发病起着重要作用，情志、生理、社会和各种生物因素相互作用，共同参与了FD的发生。全球FD患病率占世界总人口的5%-11%，我国人群患病率为18%-45% 。FD归属于中医学“痞满”范畴，以肝郁脾虚为基本病机，中医学认为反复、持久的情志刺激能影响肝气的疏泄，肝失疏泄，脾不运化，胃不受纳，引起FD的发生。课题组前期临床观察发现院内制剂平胃胶囊在治疗FD方面疗效显著，但其发挥效应属“多因微效”，是多个弱效应的整合作用，而其具体治疗的分子机制尚未阐明。目前，FD的发病尚不能用单一的效应机制解释，脑肠轴的异常是FD发病机制的共同通路。因此，本课题以脑肠轴为切入点，以肝郁脾虚型FD大鼠为研究对象，观察各组大鼠体重、饮食、病理形态学改变、胃肠电活动及胃排空和肠推进，借助ELISA、免疫组化、RT-PCR、Western-Blot等方法，观察平胃胶囊对各组大鼠脑肠肽水平表达的影响，结果发现平胃胶囊可纠正不同脑肠肽水平的分泌和表达；运用双向电泳技术筛选出不同蛋白质，结果发现平胃胶囊可不同程度的上调或下调差异蛋白，这些差异蛋白均涉及细胞骨架和形态，能量代谢及生物氧化等。故认为通过脑肠轴纠正异常脑肠肽水平的表达和分泌可能是平胃胶囊治疗肝郁脾虚型FD的机制与作用途径。