基于胃Cajal间质细胞研究厚朴 "辛-行"改善胃动力障碍的作用机制

Mangnolia officinalis is a representative of Chinese materia medica with pungent in flavour,warm in property,acting on the spleen and stomach, and it has the efficts of promoting the flow of qi and eliminating dampness. It is used to cure the symptom-complex with stagnation of qi in the spleen and stomach. So far, research on the linkages between mechanism of the efficts that Mangnolia officinalis can promote the flow of qi o and TCM function mechanism is rare. Then How to explain the connotation from the perspective of modern pharmacology.This project is one of the key answers. Mangnolia officinalis is the object of this project,and serum pharmacology, electrophysiology, immunohistochemistry, RT-PCR, and other advanced technologies are used in this project , which combines the knowledge of TCM function mechanism -chemistry- pharmacology. Gastric interstitial cells of Cajal which can coordinate smooth muscle of gastrointestinal and regulate transmission of gastrointestinal neural is the center of this project. If the project is implemented, we will explore the molecular biological mechanism of Mangnolia officinalis regulate the gastric interstitial cells of Cajal around the cell messengers - calcium channel - neurotransmitter receptor - electrophysiological nerve signals pass pathway axis in the overall - the target organ - the cellular and molecular level. Through the implementation of this project, it will verify the hypothesis that Mangnolia officinalis with pungent in flavour which can promoting flow of qi has the effict of improving gastric motility disorders because of its effect on the gastric interstitial cells of Cajal, and then it will provide new idea to research the scientific interpretation of correlation of pungent ,warm ,acting on the spleen and stomach and promoting the flow of qi in the TCM function mechanism.

厚朴具有显著的"行气燥湿"功效,其"辛温，归脾、胃经"，主治脾胃气滞诸证。迄今对厚朴发挥行气功效的作用机制及其与药性之间的关联性研究不多见。本项目拟厚朴为研究对象，采用血清药理、电生理、免疫组化、RT-PCR等先进技术，结合中药药性-化学-药理等多学科知识，以协调胃肠平滑肌运动、调控胃肠道神经信息传递的核心细胞- - 胃Cajal间质细胞为切入点，围绕细胞信使-钙通道-神经递质与受体-电生理等神经信号传递通路的研究轴，从整体-靶器官-细胞分子水平，系统探索厚朴启动胃Cajal间质细胞的分子生物学机制，以验证"厚朴通过启动胃Cajal间质细胞及其相关机制从而发挥'辛-行'特点改善胃动力障碍"的工作假说，进而为科学阐释中药药性理论中"辛-温-归脾胃经-行气"的关联性提供新的思路。

厚朴具有显著的"行气燥湿"功效,其“辛温，归脾、胃经”，主治脾胃气滞诸证。迄今对厚朴发挥行气功效的作用机制及其与药性之间的关联性研究不多见。本项目以厚朴为研究对象，采用电生理、免疫组化、RT-PCR、western-blot等先进技术，通过研究厚朴对胃Cajal间质细胞的影响，为阐释“味辛-行气”性效关联性的科学内涵提供依据。整体动物方面研究显示：厚朴水煎液及其总酚提取物能显著促进小鼠胃排空，降低胃残留率，能显著增加大鼠胃电慢波频率；对阿托品所致胃肠动力障碍大鼠，厚朴水煎液、及总酚提取物能显著增加血清中GAS、MTL、SP含量；厚朴水煎液及厚朴总酚提取物能显著增加大鼠胃窦和胃体组织Cajal间质细胞表达量；厚朴水煎液及厚朴酚类提取物能显著增加大鼠胃起搏区c-kit、SCF蛋白及胃窦部位c-kit蛋白相对表达量。原代培养细胞研究显示：厚朴含药血清、厚朴总酚含药血清显著提高Cajal间质细胞活力；厚朴含药血清、厚朴总酚含药血清能显著降低组细胞内Ca2+含量，显著降低细胞Ca2+-ATPase酶活性；厚朴含药血清组及厚朴总酚含药血清组能显著降低细胞内磷脂酶C（PLC）、三磷酸肌醇（IP3）的含量；厚朴含药血清及厚朴总酚含药血清能显著升高细胞内c-kit mRNA相对表达量，显著降低细胞内CaV1.3 mRNA、nNOS mRNA、HO-2 mRNA相对表达量；厚朴含药血清及厚朴总酚含药血清能显著升高c-kit蛋白表达，能显著降低CaV1.3、nNOS、HO-2 蛋白表达。本项目研究表明，厚朴“味辛-行气”之性效关联的科学内涵，与其通过影响胃Cajal间质细胞Ca2+通路相关指标PLC、IP3、CaV1.3、nNOS、HO-2，上调胃Cajal间质细胞中c-kit、SCF水平、增加胃Cajal间质细胞的数量、升高血清中GAS、MTL以及SP的含量，从而刺激胃电活动、收缩胃部平滑肌以及加快胃排空以促进胃动力有关。厚朴总酚类提取物可能为其发挥作用的物质基础。本项目验证了“厚朴通过启动胃Cajal间质细胞及其相关机制从而发挥‘辛-行’特点改善胃动力障碍”这一假说，这为科学阐释中药药性理论中“辛-温-归脾胃经-行气”的关联性提供新的思路，进而以促进中药药性理论的发展。