基于中医"痰“、”饮“理论的泽漆抑制慢性阻塞性肺疾病炎症的作用机制与物质基础研究

Chronic obstructive pulmonary disease(COPD) is a serious disease endangering human health. Inflammation is the core nosogenesis of COPD. Hyaluronic acid (HA) plays a major role in airway inflammation and airway remodeling. TCM believes that the so called phlegm and fluid is the key pathogenesis of COPD. HA with different concentrations and molecular weight has the difference in self-aggregation, hydrated and viscosity, which fits the clinical manifestation of TCM "phlegm and fluid " . Therefore, it is very likely that HA is one of material basis which associates COPD inflammation with TCM "phlegm and fluid ". The tast group chooses Euphorbia Helioscopia L. as the research object. The project adopts the combination of traditional water decocting and macroporous resin isolation to obtain the total extraction and some chemical fractions of Euphorbia Helioscopia L. The project adopts smoking method to duplicate the COPD pharmacological model, and screen the active fractions for the inhibition of inflammation by Euphorbia Helioscopia L.. Using immunohistochemical method and molecular biology technology, the project detects the expression change and content of HA and its receptor CD44 in rat blood, lung tissue and bronchoalveolar lavage fluid separately before and after treating with the above active fractions. Meanwhile, the project detects the change of the index including oxidation / antioxidation, protease / antiprotease , cytokine ,mediators of inflammation associated with HA and its receptor CD44,etc. And finally, with the acute toxicity test ensuring the safety of the total extraction and the active parts of Euphorbia Helioscopia L., the project attempts to reveal preliminarily that Euphorbia Helioscopia L. eliminates both phlegm and fluid with regulating HA and inhibits COPD inflammation..

慢性阻塞性肺疾病(COPD)严重危害人类健康，炎症是其核心发病机制，透明质酸（HA）在气道炎症中作用重要。中医认为“痰”“饮”是COPD的关键病因病机。浓度不同的HA，其自我聚集、水合、粘滞性的功能而不同，契合了中医“痰”“饮”不同的致病特点。因此，HA极可能是密切联系中医“痰”“饮”与COPD炎症的物质基础之一。本课题组以泽漆为研究对象，采用传统水煎煮法与大孔吸附树脂分离技术相结合，得到泽漆总提取物及若干化学部位。采用烟熏法复制COPD药效模型，筛选抑制炎症的有效部位并进行干预治疗；应用免疫组化、分子生物学技术检测干预前后大鼠血液、肺组织和肺泡灌洗液中HA及受体CD44 表达与含量变化，并检测与之相关的炎症介质、细胞因子、氧化/抗氧化、蛋白酶/抗蛋白酶等方面指标的变化。在急性毒性实验确保用药安全的前提下，揭示泽漆通过调控HA化“痰”、“饮”，抑制COPD炎症的作用机制与物质基础。

慢性阻塞性肺疾病(COPD)严重危害人类的健康。现代医学揭示，炎症是其核心发病机制，透明质酸（HA）在气道炎症中具有重要的影响。中医提出“痰”“饮”是COPD的关键病因病机。基于HA的自我聚集、水合、粘滞性的不同作用，与 “痰”“饮”不同的致病特点契合，HA极可能是密切联系中医“痰”“饮”与COPD炎症的物质基础之一。本项目以治疗COPD有确切临床疗效的泽漆为研究对象，确保用药安全基础上，通过泽漆水提取物及若干化学部位干预治疗烟熏法COPD药效模型，筛选抑制炎症的有效部位；并应用免疫组化、分子生物学技术检测干预后大鼠HA及受体CD44 的变化，并检测与之相关的炎症介质、细胞因子、氧化/抗氧化、蛋白酶/抗蛋白酶等方面指标的变化。. 经药效筛选，泽漆水提物、二氯甲烷萃取部位与乙酸乙酯萃取部位是泽漆抑制COPD炎症的有效部位，且是安全的。泽漆水提物与二氯甲烷部位疗效更优。通过对泽漆的系统化学研究，提取分离得到了26个化学成分。5个已知化合物为首次从大戟属植物中分离得到，8个已知化合物为首次从泽漆中分离得到。 . 泽漆水提物与二氯甲烷部位抑制COPD炎症的作用较为接近。其可能作用机制：通过调控HA及其受体CD44与二者的下游生物信息转导过程中的相关因子，发挥抑制COPD炎症的作用。此类相关因子主要有①细胞因子TNF-α、TGF-β1的；②蛋白酶/抗蛋白酶相关指标MMP-9、TIMP-1等；③氧化/抗氧化相关指标SO、MDA等；④中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞。而乙酸乙酯部位的作用机制：可能与降低炎症介质HsCRP、LTB4的含量，调控细胞因子TNF-α、TGF-β1有关。. 本项目初步揭示，HA及其受体CD44与二者的下游生物信息转导过程中的相关因子，极有可能是COPD“痰”“饮”的基本物质基础之一，也是相关药物的干预通路之一。这为今后泽漆治疗COPD的新药研发奠定坚实的基础。