针刺俞募穴治疗功能性便秘的EGC及ENS-ICC-SMC功能网络调整机制研究

Functional constipation (FC) is a common ailment and frequently encountered disease in department of gastroenterology, which is seriously reducing patients' quality of life. However, effect of medication treatment is not satisfactory and a better solution is badly needed. Acupuncture has been sufficiently proved effective in treating FC by clinical practice. FC is among 43 diseases recommended by WHO to be treated by acupuncture. Back-Shu and Front-Mu acupoints is a classical acupuncture approach for treating FC, but the mechanism behind has not been explained essentially. Recent studies indicate that the key point in pathogenesis of FC is decrease of EGC and unbalance of ENS-ICC-SMC functional network. Exploring the peripheral mechanism of acupuncture is probably a way to reach a breakthrough. Therefore, this project based on acupuncture Front-mu and Back-shu, a classic combination of acupuncture points, takes the functional constipated model mice as research subjects, further explains the scientific content of acupuncture in treating gastrointestinal disorders. The project starts with EGC, ENS-ICC-SMC network, and GDNF-PI3K-Akt signal transduction pathway, which are important factors effecting on gastrointestinal motility, then utilizes techniques such as immunohistochemistry, situ hybridization, quantitative image analysis performed with computer, fluorescence quota PCR and gene knockout to prove the regulation effect of acupuncture on gastrointestinal motility and functional network of FC model mice pros and cons.

功能性便秘是消化系统的常见病、多发病，严重影响患者生活质量，药物治疗不理想，是目前亟待解决的临床热点问题。针灸治疗功能性便秘疗效确切，证据充分，是WHO推荐针灸治疗的43种疾病之一，其中大肠俞募穴是最经典、最常用的取穴方法，然而俞募穴治疗该病的作用机制尚未阐释清楚。最新研究表明EGC减少进而引起ENS-ICC-SMC功能网络失调是功能性便秘发病的关键环节，从外周机制入手可能是阐释作用机制的突破口。故本项目以针灸俞募穴理论为指导，以功能性便秘模型小鼠为研究对象，从影响胃肠动力的关键环节EGC、ENS-ICC-SMC功能网络、以及密切相关的GDNF-PI3K-Akt信号转导通路入手，运用免疫组化、原位杂交结合计算机图像分析、荧光定量PCR等方法，并采用基因敲除技术，从正、反两面证实针刺俞募穴对模型小鼠肠道动力、肠道功能网络的调整作用，拟深入阐释俞募穴治疗胃肠疾病这一针灸经典用穴规律的科学内涵。

功能性便秘是消化系统的常见病、多发病，严重影响患者生活质量，药物治疗不理想，是目前亟待解决的临床热点问题。针灸治疗功能性便秘疗效确切，证据充分，是WHO推荐针灸治疗的43种疾病之一，其中大肠俞募穴是最经典、最常用的取穴方法，然而俞募穴治疗该病的作用机制尚未阐释清楚。最新研究表明EGC减少进而引起 ENS-ICC-SMC功能网络失调是功能性便秘发病的关键环节，从外周机制入手可能是阐释作用机制的突破口。. 本项目以针灸俞募穴理论为指导，以功能性便秘模型小鼠为研究对象，从影响胃肠动力的关键环节 EGC、ENS-ICC-SMC功能网络、以及密切相关的 GDNF-PI3K-Akt 信号转导通路入手，运用免疫组化、原位杂交结合计算机图像分析、荧光定量 PCR 等方法，并采用基因敲除技术，从正、反两面证实针刺俞募穴对模型小鼠肠道动力、肠道功能网络的调整作用，拟深入阐释俞募穴治疗胃肠疾病这一针灸经典用穴规律的科学内涵。. 本项目分别从针刺俞募穴对不同状态小鼠胃肠传输功能影响、针刺俞募穴对功能性便秘小鼠EGC及ENS-ICC-SMC功能网络的调整机制和针刺俞募穴对功能性便秘小鼠GDNF-PI3K-AKT信号转导通路影响三个方面的实验来进行研究。. 实验结果表明：1.针刺刺激可加快功能性便秘小鼠胃排空和小肠推进率，提高结肠平滑肌的收缩张力，进而增强便秘小鼠胃肠道传输功能。正常小鼠针刺后各指标变化不明显，提示在健康状态下，针刺对胃肠传输功能的作用不明显。 2.针刺可能通过EGC细胞的作用来改善功能性便秘小鼠的排便情况。针刺能够降低功能性便秘平滑肌细胞的凋亡，改善平滑肌细胞结构，提高ICC细胞及肌间神经丛的数量，从而恢复ENS-ICC-SMC功能网络。3.针刺大肠俞募穴可能通过促使EGC细胞分泌产生GDNF，调控PI3K-Akt信号通路，抑制mTOR的表达，进而减少细胞凋亡，改善结肠组织形态及胃肠传输功能。