骨桥蛋白在慢性阻塞性肺疾病小鼠肺组织中的表达以及细辛脑的干预作用

Effect of asarone on osteopontin expression in the lung tissue of chronic obstructive pulmonary diseases model mice

目的初步研究骨桥蛋白(OPN)在慢性阻塞性肺疾病(COPD)模型小鼠肺组织中的表达以及细辛脑对其表达的干预作用及其治疗意义。方法将40只BALB/C雌性小鼠随机分为对照组(A组)、COPD组(B组)、细辛脑组(C组)和富露施组(D组),每组10只。采用鼻腔滴入脂多糖(LPS)加熏香烟的方法建立COPD小鼠模型。运用HE染色对肺组织病变程度进行观察,评价COPD小鼠模型;Western blot方法检测肺组织中OPN的表达。计数血中白细胞;酶联免疫吸附试验(ELISA)法检测肺泡灌洗液中OPN的含量;免疫组化分析肺组织切片中OPN的表达。数据结果采用SPSS 17.0软件进行分析,两组均数比较采用两独立样本t检验,多组均数比较采用One-way ANOVA方差分析。结果 HE染色显示:COPD组与对照组相比,支气管管壁增厚,炎性细胞浸润明显,肺泡扩大甚至断裂。细辛脑组和富露施组支气管管壁厚度,炎性细胞浸润肺泡扩大程度较COPD组明显减轻;ELISA检测显示:与对照组相比,COPD组肺组织中的OPN表达明显增高(t=12.27,P=0.003),与COPD组相比,细辛脑和富露施组肺组织中的OPN表达明显减少(F=531.22,P<0.05)。血中白细胞计数:COPD组明显高于对照组(t=7.32,P<0.05),细辛脑组和富露施组白细胞计数明显少于COPD组(F=113.53,P=0.001);免疫组织化学的结果显示:与对照组比,COPD组OPN表达明显增加(t=5.67,P<0.05);与COPD组相比,细辛脑组和富露施组OPN的表达量明显减少(F=172.11,P=0.001)。结论 OPN可能是COPD疾病中重要的炎性介质,在气道炎症的发展和维持中起重要作用,细辛脑可以通过干扰OPN的生成,减弱COPD模型小鼠炎症的进展。

Objective To investigate the effect of asarone on osteopontin expression in the lung tissue of chronic obstructive pulmonary diseases model mice, providing therapeutic significance. Methods 40 female mouse of BALB/C were randomly divided into control group, COPD group, asarone group and fluimucil group. COPD mice model was established by using the method of intranasal instillation of lipopolysaccharide(LPS) and smoked cigarettes. To observe the degree of pathological changes of lung tissue and evaluate the COPD mice model by using HE staining. The level of OPN in the mice tissue were determined by Western blot; The expression of OPN in lung tissue was analyzed by immunohistochemistry. Results The HE staining showed compared with the control group, the thickness of bronchial wall and degree of inflammation cell infiltration and damage of alveolar of COPD group were more evident. While the thickness of bronchial wall and degree of inflammation cell infiltration and damage of alveolar of asarone and fluimucil group decreased significantly. The Western blot and ELISA showed the level of OPN in mice lung tissue increased in the COPD group compared with the control group(P<0.05), but the level of OPN in mice lung tissue decreased more significantly in the asarone and fluimucil group compared with COPD group(P<0.05). The immunohistochemical showed that compared with the COPD group, the expression of OPN in lung tissue decreased significantly in the asarone and fluimucil group(P<0.05). Conclusions The expression of OPN in COPD may be an important inflammatory mediator, which plays an important role in the development and maintenance of the airway inflammation. Asarone can attenuate the inflammation of COPD model mice by interfering the generation of OPN.