基于HCN离子通道研究半夏泻心汤对功能性消化不良胃肠运动的干预机制

Gastrointestinal motility disorder is the major pathophysiological basis of occurrence and development of functional dyspepsia（FD）, and this was mainly due to abnormal pacing of gastrointestinal smooth muscle. Chinese medicine believes that FD is a " distention and fullness " category. Han Dynasty "Shanhan lun" revealed representative prescription of distention and fullness is Banxiaxiexin decoction which has the effect on treatment of FD. Previous study found that the therapeutic treatment of Banxiaxiexin Decoction is to regulate gastrointestinal motility , but the targets are unknown. HCN channels has pacing, regulating and contracting function, exists the gastrointestinal mucosa, smooth muscle and ICC and participates in the gastrointestinal smooth muscle contraction and regulation of pacing. Assuming topics BanxiaxiexinTang intervention by HCN gastrointestinal motility. Utilized modern biological technology to measure plasma GAS, MTL, SS, VIP, stomach residual rate ratio and small intestine propulsion ratio, gastrointestinal muscle tension, HCN and co-expression of c-kit, gastrointestinal neurotransmitters SP, CGRP, VIP and HCN co-expression , HCN channel currents ，protein expression of HCN and mRNA expression in vivo and in vitro. From the level of organs, tissues , and cellular and molecular, this research is to explore the mechanism of Banxiaxiexin Decoction and to reveal targets, providing a theoretical basis for the expansion of the clinical application.

胃肠运动障碍是FD发生、发展的主要病理生理基础，且该障碍主要源于胃肠平滑肌起搏异常。中医认为FD属“痞满”范畴。汉代《伤寒论》主治寒热互结痞证代表方半夏泻心汤，治疗FD临床疗效确切。我们既往研究发现半夏泻心汤治疗作用主要是调节胃肠运动，但作用靶点不明。ICC是胃肠起搏细胞，同时，具有起搏、调节收缩功能的超极化激活环核苷酸门控阳离子通道(HCN)在胃肠粘膜、平滑肌、ICC上也普遍存在，并参与胃肠平滑肌收缩起搏和调节。课题假设半夏泻心汤通过HCN干预胃肠运动。运用现代生物学技术，测血浆GAS、MTL、SS、VIP，胃残留率和小肠推进比，胃肠肌条张力，HCN和c-kit共表达，胃肠递质SP、CGRP、VIP和HCN共表达，HCN通道电流，在体和体外相结合测HCN的蛋白表达、mRNA表达，从整体、器官、组织和细胞分子水平，进一步探讨半夏泻心汤作用机制，揭示其作用靶点，为扩大其临床应用提供理论依据。

本研究探讨了半夏泻心汤对功能消化不良的治疗作用及机制。主要是基于HCN离子通道研究了半夏泻心汤对功能性消化不良大鼠胃肠运动的调节研究，内容与结果如下：①系统地研究了功能性消化不良发病时外周胃肠激素、局部胃肠激素、胃肠起搏细胞、HCN离子通道与胃肠运动的关系。②HCN1和HCN2离子通道在胃肠间质细胞ICC细胞的电学活动中所起的重要作用，HCN1和HCN2离子通道表达水平一定程度上反映了ICC细胞信息传递功能。③HCN1和HCN2离子通道调节胃肠道运动的起搏，并提示HCN1和HCN2离子通道异常与功能性消化不良发病有关。④胃肠激素与HCN1和HCN2离子通道的共表达，提示胃肠激素通过HCN1和HCN2离子通道实现对胃肠运动的调节。⑤从动物模型整体、器官、组织、细胞到分子水平系统证实了半夏泻心汤能有效治疗功能性消化不良。⑥半夏泻心汤能调节HCN1和HCN2离子通道的表达和离子通道电流，其治疗功能性消化不良一定程度上是通过调节HCN1和HCN2离子通道的表达和离子通道电流实现的。提示HCN离子通道是半夏泻心汤的作用靶点。本课题体内实验与体外实验相结合验证其作用及作用机制，从动物模型整体、器官、组织、细胞到分子水平系统研究，验证了预定设想，完成了预期计划。