基于热敏通道间的交叉脱敏探讨理中丸、小建中汤和吴茱萸汤温中散寒功效的共同作用机制

Warming the middle Jiao for dispelling the cold is a method to treat the coldness in the middle Jiao, which is one of the most important content in Warming formulas. However, its pharmacological principle is not clear, especially based on the cellular and molecular biological meaning. A recent research states that the Thermal-TRP Ion Channel on harmful receptors of visceral sensory nerves not only can feel the temperature, but also be able to mediate either acute or chronic inflammatory pain. In addition, it is also the functional target of many pungent herbs that can desensitize in each others. It is the basis of Cold Remedies Heat in cellular and molecular biological point of view. This study will proof that the three formulas, LiZhongWan, XiaoJianZhongTang, and WuZhuYuTang, having the same pharmacological effect on the Thermal-TRP Ion Channel by setting the TRPV1/TRPA1 on the organs (Stomach, intestines, liver, gallbladder, and pancreases) in the middle Jiao as the object of the study, a laboratory rat with high level of TRPV1/TRPA1 as a model who is suffering the pain in organs caused by inflammation, and TRPV1/TRPA1 receptor agonist as the test items, such as the GanJiang/ShengJiang, GuiZhi, WuZhuYu in the three formulas to reveal the principle of warming the middle Jiao for dispelling the cold. This will be another support in TCM treatment principle of Hot Remedies Coldness, as well as an important basis or guide for TCM doctors to choose better formula or herbs to treat.

温中祛寒剂的代表方理中丸、小建中汤和吴茱萸汤（温中三方）都有温中散寒的功效，在治疗中焦寒证时常可互换使用，因此可能有共同的作用机制。现代研究表明，分布在感觉神经伤害性感受器上的热敏通道既可感知寒热，又能介导多种急慢性疼痛，还是许多辛味中药的作用靶点。激动剂在热敏通道之间可以交叉脱敏，这可能是“寒者热之”的药理基础。据此我们推测温中三方都可以激活热通道TRPV1，产生温热效应；而TRPV1通道的活化对TRPA1产生交叉脱敏，阻断寒痛信号的传导，最终达到温中散寒的作用。本项目采用消化道炎症小鼠模型，以温中三方进行干预，采用分子生物学方法对消化道组织和背根神经TRPV1/TRPA1的mRNA和其调控因子的表达进行检测。同时使用电生理方法在细胞上筛选和比较三方激动剂药效。以此证明温中三方在细胞和分子水平上有共同的药效基础。本研究将为阐明方剂功效的科学内涵提供实验依据，为临床合理用方提供指南。

温中祛寒剂的代表方理中丸、小建中汤和吴茱萸汤（温中三方）都有温中散寒的功效，在治疗中焦寒证时常可互换使用，因此可能有共同的作用机制。现代研究表明，分布在感觉神经伤害性感受器上的热敏通道既可以感知寒热，又能介导多种急慢性疼痛，还是许多辛味中药的作用靶点。激动剂在热敏通道之间可以交叉脱敏，这可能是“寒者热之”的药理基础。据此，我们推测温中三方都可以激活热敏通道TRPV1，产生温热效应；而TRPV1通道的活化对TRPA1产生交叉脱敏，阻断寒痛信号的传导，最终达到温中散寒的作用。本项目采用消化道炎症小鼠模型，以温中三方进行干预，采用分子生物学方法对消化道组织和背根神经TRPV1/TRPA1的mRNA和其调控因子的表达进行检测，同时使用电生理方法在细胞上筛选和比较三方激动剂药效，以此证明温中三方在细胞和分子水平上有共同的药效基础。研究结果显示，运用膜片钳技术，在细胞和分子水平验证和发现了干姜、生姜、吴茱萸和桂枝是热敏通道TRPV1/TRPA1的激动剂。在转染TRPV1的HEK293细胞上使用膜片钳技术验证了吴茱萸碱、10姜酚是TRPV1激动剂，配伍使用增效。在动物内脏疼痛行为学模型中证明，温中三方均有明显的止痛作用，而且是通过热敏通道TRPA1/TRPV1实现的。