基于Toll样受体信号通路“危险模式”效应理论解析温中益气健脾法治疗Hp相关性溃疡的分子机制

The Helicobacter pylori-related peptic ulcer (HpPU) is a frequently-occurring, multi-site, and chronic disease of the digestive system. Impairment of the gastrointestinal mucosal barrier is the main pathological link leading to HpPU. Activation of toll like receptor 2 (TLR-2) signaling pathway can trigger the "danger model" effect, release cytokines to protect the mucosal barrier, and promote ulcer healing. Chinese medicine believes that the therapy of warming the Middle, tonifying Qi, and invigorating the spleen contributes to mucosal barrier reconstruction, muscle renewal, and ulcer repair. In the preliminary experiments, we found that Huang-Qi-Jian-Zhong-Tang (HQJZT) could promote the repair of gastric mucosal injury and increase the protein expression of TLR-2 in HpPU rats. Therefore, we propose the hypothesis that HQJZT can repair mucosal barrier damage via regulating “danger model” effect triggered by TLR-2 signaling pathway, thus promoting ulcer healing to treat HpPU. To verify this hypothesis, we intend to carry out the in vitro and in vivo experiments and clinical research, by using techniques such as RNA interference, electron microscopy, suspension chip, etc. The quality of mucosal barrier healing will be used as the main evaluation index. The effect and possible mechanism of "danger model" effect triggered by TLR-2 signaling pathway for HQJZT in treating HpPU are going to be systematically investigated from the morphology and function perspectives, animal-cell-molecular levels, and basic and clinical aspects. The aim of this project is to provide scientific basis for the clinical application of integrated traditional Chinese and western medicine in the treatment of HpPU.

Hp相关性溃疡(HpPU)是消化系统的一种多发性慢性疾病。胃肠黏膜屏障损伤是导致HpPU的主要病理环节。Toll样受体2(TLR-2)信号通路激活可触发“危险模式”效应，释放黏膜屏障保护因子，促进溃疡愈合。中医学认为，温中益气健脾治法有助于黏膜屏障重建、肌肉续接、溃疡修复。我们在发现黄芪建中汤可促进HpPU大鼠胃黏膜组织损伤修复及上调TLR-2蛋白表达的基础上，提出“黄芪建中汤通过调控TLR-2信号通路‘危险模式’效应修复黏膜屏障损伤，促进溃疡愈合，治疗HpPU”的假说。为验证该假说，本项目拟开展体内外实验和临床研究，采用RNA干扰、电镜、悬液芯片等技术，以黏膜屏障愈合质量为评价指标，从形态和功能角度、动物-细胞-分子水平、基础与临床层面系统考察黄芪建中汤治疗HpPU的效果及其通过TLR-2信号通路“危险模式”效应的可能机制，为中西医结合治疗HpPU在临床的推广应用提供科学依据。

消化性溃疡（peptic ulcer, PU）是一种多致病因素引起的以胃和十二指肠溃疡为病理特征的慢性疾病，通常把幽门螺杆菌（Helicobacter pylori, Hp）感染引发的PU称为Hp相关性溃疡（HpPU）。HpPU难治愈、易复发，且可出现大出血、穿孔、癌变等严重并发症，因此本病的防治一直是临床研究的热点和难点问题。HpPU发病机制的现代理念认为：胃肠黏膜屏障损害是HpPU发病的基本原因，溃疡的发生是黏膜屏障被破坏的结果。Toll样受体2（TLR-2）是机体免疫系统的辅助受体，广泛分布于胃肠黏膜，TLR-2信号传导通路被阻断可开启“危险模式”效应，释放多种黏膜保护因子，以维持胃肠黏膜屏障完整性。TLR-2是修复黏膜屏障损伤，促进溃疡愈合，治疗HpPU的重要靶点。中医理论认为脾胃虚寒是HpPU发病与转归的根本内因。温中益气健脾法是促进HpPU愈合、预防复发的关键治法。本项目通过构建HpPU大鼠模型，发现温中益气健脾法代表方黄芪建中汤和理中汤均具有较好的抗大鼠胃溃疡和十二指肠溃疡的效应，其机制与抑制TLR-2/MyD88信号通路，开启“危险模式”效应，增强黏膜屏障功能，释放多种黏膜保护因子有关。通过采用Tips划痕法构建GES-1细胞损伤模型，发现黄芪建中汤和理中汤含药血清均能促进GES-1细胞迁移和增殖，其机制与阻断TLR-2/MyD88信号通路有关。通过对1781例HpPU临床患者的meta分析发现，黄芪建中汤联用常规西药治疗HpPU的有效性（临床治愈率、临床总有效率、Hp根除率、复发率）和安全性（不良反应发生率）均优于单用常规西药治疗。本项目将温中益气健脾治法方药的研究深入到细胞内信号通路及相关基因和蛋白表达水平，为临床提高HpPU患者愈合质量及抑制溃疡复发提供了治疗思路和方法，为中西医结合治疗HpPU在临床的推广应用提供了科学依据。