中药高良姜抗胃溃疡物质基础及作用机制研究

The rhizomes of Alpinia officinarum have the efficacy of warming middle-jiao to dispel cold and promoting qi to relieve pains. They are commonly used in clinic for the treatment of gastric ulcer and have been shown obvious effect. However, the effective substances and mechanism of anti-gastric ulcer activity are still not clear. Our previous research has determined the effective fraction with anti-gastric ulcer effect of the rhizomes of A. officinarum, and found that flavonoids and diarylheptanoids were the main active components, which may act on NF kappa B and the capsaicin receptor (TRPV1) to exert the anti-inflammatory and analgesic effects. On the basis of the previous work, this project intends to divide the effective fraction into flavonoid and diarylheptanoid sub-fractions. The two sub-fractions will undergo systematic chemical isolation and the then the structures of isolated compounds will be elucidated and identified. The active ingredients will be clear and then the effective substances of the anti gastric ulcer will be clarified further by the investigation of animal models of gastric mucosal injury and gastric ulcer. The mechanism of the anti-gastric ulcer of the effective fraction, effective groups and active compounds will be explored by molecular biology techniques from two aspects: action on the “inflammation” of gastric ulcer mediated by NF-κB pathway and “pain” of gastric ulcer mediated by TRPV1. Through the study of this project, we will illustrate fundamentally the scientific connotation of “warming middle-jiao” and “relieving pain” of the rhizomes of A. officinarum, and provide scientific basis for the rational application in clinic and further utilization and development of this crude drug. The research will also provide references for the investigation of Chinese materia medica with anti-gastric ulcer activity from multi-targets and multi-level aspects.

中药高良姜具有温中散寒、行气止痛之功效，临床常用于治疗胃溃疡效果显著，但其抗胃溃疡的药效物质基础及作用机制尚不明确。课题组前期确定高良姜抗胃溃疡有效部位，并发现黄酮和二芳基庚烷部位为其主要活性组分，且可能作用于NF-κB，辣椒素受体（TRPV1）发挥抗炎、止痛功效。本项目拟在前期工作的基础上，对有效部位进行拆分得到黄酮和二芳基庚烷组分，并对这两个组分进行系统的化学成分分离，确定化合物的化学结构，并结合胃粘膜损伤模型、胃溃疡动物模型研究明确活性成分，阐明其抗胃溃疡的药效物质基础；进一步采用分子生物学技术探索高良姜有效部位、有效组分、有效成分通过作用于NF-κB通路介导的胃溃疡“炎性”和TRPV1介导的胃溃疡“疼痛”两个角度治疗胃溃疡的作用机制。通过本项目的研究，基本阐明高良姜“温中”、“止痛”的科学内涵，为高良姜的临床合理应用与开发提供科学依据，为中药抗胃溃疡多靶点、多层次研究提供参考。

高良姜为姜科山姜属植物高良姜（Alpinia officinarum Hance）的干燥根茎，具温中散寒、行气止痛之效，其核心功效之一是治疗胃脘痛，在临床上具有明确的应用历史。课题组在前期已明确高良姜抗胃溃疡活性,并建立了有效部位的分离与制备方法，课题组将其有效部位按化学成分类型进行拆分得到黄酮组分与二芳基庚烷组分，并对各组分进行分析（黄酮组分主要包含3个黄酮类化合物，庚烷组分包含9个庚烷类化合物）。后通过体内（乙醇诱导小鼠急性胃溃疡、乙酸诱导大鼠慢性胃溃疡）和体外（乙醇诱导人胃黏膜上皮细胞（GES-1）损伤）实验对高良姜的有效组分（二芳基庚烷组分和黄酮组分）及其中的有效单体成分（高良姜素、山柰素等）进行抗胃溃疡的活性研究和机制分析。发现这些组分和单体能对急慢性胃溃疡损伤模型均具有显著的改善作用，其作用效价相似或优于阳性对照药雷尼替丁。体外实验结果提示，黄酮组分能够抑制乙醇诱导细胞线粒体途径中的凋亡基因Bax、Caspase-3表达增加及抗凋亡基因Bcl-2的表达降低并通过增加Egf及其受体Egfr蛋白的表达提高细胞活性、迁移能力促进细胞自愈及溃疡修复能力。乙醇诱导急性胃溃疡实验证明，黄酮组分通过NF-κB/COX-2和iNOS/NO通路、二芳基庚烷组分通过TLR4/NF-κB和TRPV1/CGRP-SP通路发挥抗胃溃疡的作用。此外发现，黄酮组分能够改善内皮素相关性炎症治疗慢性胃溃疡，在高通量测序探讨慢性胃溃疡潜在发病机制和黄酮组分在胃溃疡治疗中的作用并确立靶点途径中发现，黄酮组分与PI3K-Akt、Toll样受体、炎性介质TRP信号通路有关，从而在加速溃疡愈合修复，抑制炎性神经疼痛传递和避免自身功能免疫调节障碍机制中起到胃溃疡治疗作用。以上实验结果为阐述高良姜传统功效（温中、止痛）的科学内涵提供了重要的依据，为高良姜的化学成分研究及临床合理应用和开发提供重要的保障。此外，在进行项目研究的同时，开展了关于高良姜甘油三酯类及多糖类成分的分析与表征，丰富了高良姜的物质基础研究，为进一步的研究打下基础。