基于HCN离子通道研究滋阴行气法对功能性便秘大鼠结肠运动的干预机制

Researches home and abroad indicate that HCN is widely distributed in myenteric plexus of the gastrointestinal tract, and plays an important role in the regulation of gastrointestinal dynamics, particularly the expression and distribution trends of HCN1、HCN2 are in highly consistent with the ICCs. Our research shows that c-Kit positive ICC cells have antigen expression of HCN channels, which is consistent with the research at home and abroad； Previous clinical studies have shown that Changrunfang decoction had a 90% short-term effect in treating functional constipation, the effect rate was 67.5% after 3 months follow-up; Changrunfang decoction can promote peristalsis, significantly improve constipation symptoms ,which may be associated with activation of colonic mucosa on the ICCs HCN. To further explore the pathogenesis of FC, clarify the pharmacological mechanism of Ziyinxingqi for FC, We make constipation big rat animal model with compound diphenoxylate, observe the contraction amplitude of colon smooth muscle article of constipation big rat away from body, and the expression of colon c-Kit, and SCF, and HCN1, and HCN2 and the, and the impact on channel current of HCN1 and HCN2 on colon Cajal. We will study the therapeutic effect of Ziyinxingqi on rats with FC, clarify its mechanism for the treatment of FC,We hope to provide new ideas and methods for prevention and treatment of constipation.

国内外研究表明HCN广泛分布于胃肠道肌间神经丛，对胃肠道的动力调节方面起着重要的作用，特别是HCN1、HCN2在结肠表达及分布变化趋势与ICCs高度一致。项目组前期研究表明c-kit阳性的ICC细胞上具有HCN通道抗原的阳性表达，与国内外研究一致;前期临床研究表明中药复方肠润方治疗FC有效率达90.0%，随访3个月有效率67.5%。肠润方能够促进肠蠕动，显著改善便秘症状，可能与其激活结肠黏膜ICCs上的HCN通道，促进肠道运动有密切关系。为进一步探索FC的发病机制，阐明滋阴行气法治疗FC的药理机制,以复方地芬诺脂制作便秘大鼠模型，通过观察肠润方对便秘大鼠离体结肠平滑肌条收缩振幅、结肠c-kit、SCF、HCN（1，2）蛋白和mRNA表达及对结肠Cajal间质细胞的HCN（1，2）通道电流的影响，进行滋阴行气法对FC大鼠的治疗作用的研究，阐明其治疗FC的机制，为便秘的防治提供新的思路和方法。

国内外研究表明超极化激活环核苷酸门控的阳离子通道(HCN)广泛分布于胃肠道肌间神经丛，对胃肠道的动力调节方面起着重要的作用，特别是HCN1、HCN2在结肠表达及分布变化趋势与结肠Cajal间质细胞（ICC）高度一致。项目组前期研究表明c-kit阳性的ICC细胞上具有HCN通道抗原的阳性表达，与国内外研究一致；前期临床研究表明中药复方肠润方治疗FC有效率达90.0%，随访3个月有效率67.5%。肠润方能够促进肠蠕动，显著改善便秘症状，可能与其激活结肠黏膜ICCs上的HCN通道，促进肠道运动有密切关系。本项目在前期工作的基础上,以复方地芬诺脂制作便秘大鼠模型，通过观察肠润方对便秘大鼠离体结肠平滑肌条收缩振幅、结肠c-kit、SCF、HCN蛋白和mRNA表达及对结肠Cajal间质细胞的HCN通道电流的影响，深入探讨滋阴行气-肠润方治疗FC的作用机制。本项目研究结果显示结肠ICCs的缺失和功能受损导致的胃肠动力障碍是功能性便秘的重要发病机制， HCN是ICC产生起搏电流及调控胃肠道神经递质的重要通道，肠润方可介导SCF/C-kit信号通路恢复ICC的数量和功能并提高HCN的表达治疗功能性便秘。此研究为进一步明确FC的可能发病机制提供一种思路，为FC的防治提供新靶点，同时丰富“滋阴行气”治疗FC的现代生物学内涵，为中医药防治便秘提供新思路。