腺苷衍生化合物WS070117M1对小鼠慢性阻塞性肺疾病的影响及其抗炎机制

Effect of WS070117M1 on chronic obstructive pulmonary disease in mice and the underling mechanisms of anti-inflammation

为了研究虫草类化合物WS070117M1[N6-(3-羟基苯基)腺苷]对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)的影响及与腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)激活相关的作用机制,以烟熏加脂多糖(lipopolysaccharide,LPS)诱导COPD小鼠实验模型,观察WS070117M1抗COPD作用;ELISA法检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中细胞因子IL-1β、IL-6、IL-8及TGF-β1水平;采用细胞分类计数仪进行BALF中炎性细胞计数;苏木素-伊红(HE)染色观察肺组织病理变化;免疫组织化学法检测肺组织p AMPKα[phospho-AMPKα(T172)]表达,ELISA法检测WS070117M1对香烟烟雾凝集物(CSC)刺激佛波酯诱导的THP-I细胞IL-8生成的影响。结果显示,WS070117M1干预抑制COPD小鼠BALF中细胞因子IL-8、IL-6、IL-1β及TGF-β1水平,减少肺部气管及血管周围炎症细胞浸润,缓解支气管上皮增生等肺部病理改变,增加肺部p AMPKα表达。WS070117M1体外可抑制CSC刺激的IL-8生成,对烟熏加LPS诱导的COPD小鼠气道炎症有抑制作用,其机制可能与其对肺部AMPK激活有关。

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-（3-hydroxylaniline） adenosine （WS070117M1） on cigarette smoke plus LPS （lipopolysaccharide）-induced chronic obstructive pulmonary disease （COPD） in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid （BALF） were harvested and IL-1β, IL-6, IL-8 and TGF-1β levels were measured by ELISA （enzyme-linked immunesorbent assay）. The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer＇s hematoxylin and eosin for histopathologic examination, pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKa phosphorylation in phorbol-12- myristate-13-acetate （PMA） differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-fll levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M 1 may alleviate the airway inflammation by activating AMPK in the lung tissue.