基于气道黏液高分泌研究COPD“阴虚痰饮”生成机制及中药的干预作用

Chronic obstructive pulmonary disease (COPD) is a major chronic diseases globally, which is the fourth cause of death currently. Airway mucus hypersecretion is a important pathophysiological characteristic of this disease, but there were deficiency in evidence of it. Inflammation directly mediates airway mucus hypersecretion with involvement of mucoprotein and aquaporins. Although syndrome type of Yin deficiency and phlegm retention is common in COPD, but its mechanism has not been reported yet. Yin-deficiency is seemingly incompatible with phlegm retention, which makes it difficult in treating. Since Yin-deficiency syndrome bears on inflammatory cytokines, and phlegm retention is associated with airway mucus hypersecretion, the mechanism of Yin deficiency and phlegm retention in COPD should be airway mucus hypersecretion due to inflammatory cytokine. Rat models of Yin deficiency-phlegm retention syndrome in COPD are induced by intratracheal lipopolysaccharide combined with fumigation and adrenal cortical hormone.Theose models and bronchial epithelial cells are observed to study the effects of inflammatory cytokine on contents and mRNA expression of mucoprotein and aquaporins protein, through which to discuss the mechanism of Yin deficiency-phlegm retention syndrome in COPD. The effects of Jin shui liu jun decoction and its components on bronchial epithelial cells are also observed to exlpore the mechanism. The research will provide scientific foundation for the clinical treatment of high mucus secretion in COPD with Yin deficiency and phlegm retention syndrome.

慢性阻塞性肺病是一种全球范围内的主要慢性疾病，列居第4位致死病因。气道黏液高分泌是其重要病理生理特征，但对于其在COPD致病机制中的研究仍十分缺乏。炎症直接介导了气道黏液高分泌的发生，而黏蛋白、水通道蛋白则与黏液高分泌密切相关。阴虚痰饮为COPD常见证型，其产生机制尚无报道。我们认为阴虚与痰饮的矛盾性导致治疗困难。阴虚证实质与炎症细胞因子有关，而痰饮与气道黏液相关，COPD阴虚痰饮机制为炎症细胞因子致气道黏液高分泌。拟以气道内注入脂多糖+烟熏+肾上腺皮质激素建立的COPD大鼠阴虚痰饮证模型和脂多糖刺激支气管上皮细胞为研究对象，观察炎症细胞因子对黏蛋白、水通道蛋白蛋白含量与mRNA表达影响，从气道黏液高分泌探讨阴虚痰饮生成机制；探讨金水六君煎及拆方对支气管上皮细胞的影响，阐明其作用机制。本研究旨在为COPD阴虚痰饮证高黏液分泌的治疗提供科学依据。

“阴虚痰饮”为慢性阻塞性肺疾病（COPD）临床常见证型，《景岳全书》提出阴虚痰饮用金水六君煎治疗。为探讨COPD“阴虚痰饮”生成机制及金水六君煎作用机理，我们体内实验采用烟熏+气道内脂多糖滴入联合地塞米松肌注建立COPD大鼠“阴虚痰饮”病证结合模型，金水六君煎及其拆方以方测证验证模型属性，从炎症细胞因子与黏液高分泌等方面探讨COPD“阴虚痰饮”生成机制及金水六君煎作用机理。体外实验采用人中性粒弹性蛋白酶诱导A549黏液高分泌细胞模型，金水六君煎及其拆方含药血清进行干预，从炎症细胞因子与黏液高分泌等方面探讨金水六君煎治疗气道黏液高分泌的可能机制。研究结果发现金水六君煎中养阴成分（熟地、当归）能拮抗糖皮质激素导致的大鼠下丘脑-垂体-肾上腺皮质轴功能抑制、环核苷酸代谢失调及Na+-K+-ATP酶活性降低。体内及体外模型均存在气道黏液高分泌相关指标变化，即黏蛋白5AC、5B（MUC5AC、MUC5B）表达明显增多，水通道蛋白5（AQP5）表达明显减少。金水六君煎体内及体外研究均能明显抑制病理状态下大鼠肺组织及A549细胞MUC5AC、MUC5B表达，促进AQP5表达。由于地塞米松对大鼠肺功能、炎症细胞因子及黏液高分泌等相关指标均存在干预作用，其诱导造模时大鼠易出现肾阴虚与肾阳虚并存表现，因此，COPD“阴虚痰饮”病证结合模型构建以及炎症细胞因子与黏液高分泌之间关系探讨未能达到满意目标。