细辛脑联合布地奈德干预哮喘小鼠气道重塑以及炎症因子的表达

Intervention of combination of asarone and budesonide on airway remodeling and expression of inflammatory factors in asthmatic mice

目的探讨骨桥蛋白(OPN)和血管内皮生长因子(VEGF)在哮喘小鼠模型中的表达和意义以及布地奈德联合细辛脑雾化吸入对模型小鼠气道重塑、细胞因子OPN和VEGF的干预作用。方法将40只BALB/C雌性小鼠随机分为对照组、哮喘组、布地奈德组和细辛脑联合布地奈德组,每组10只。采用鸡卵白蛋白(OVA)诱导致敏建立哮喘模型。HE染色观察小鼠肺组织病理改变,运用医学图像分析软件测定支气管管腔内周长(Pbm)、支气管管壁面积(WAt)、支气管平滑肌面积(WAm)、支气管平滑肌细胞计数(N),将WAt、WAm、N用Pbm标准化;酶联免疫吸附法检测小鼠肺泡灌洗液(BALF)中OPN和VEGF的含量。Western-blot分析小鼠肺组织中OPN和VEGF的含量。数据采用单因素方差分析(One-Way ANOVA),多组间两两比较采用LSD-t法,P<0.05为差异有统计学意义。结果哮喘组BALF中OPN和VEGF水平[(389.1±10.9)ng/ml、(62.3±10.3)pg/ml]高于布地奈德[(110.4±8.6)ng/ml、(38.1±7.2)pg/ml]和细辛脑联合布地奈德组[(97.6±7.9)ng/ml、(30.1±6.8)pg/ml],差异具有统计学意义(均P<0.05)。western-blot结果显示细辛脑联合布地奈德组小鼠肺组织中OPN和VEGF的阳性表达低于哮喘组和布地奈德组,差异有统计学意义(均P<0.05)。图像分析显示:哮喘组WAt/Pbm、WAm/Pbm分别高于C组(均P<0.05),细辛脑联合布地奈德组WAt/Pbm、WAm/Pbm及N/Pbm低于布地奈德组(均P<0.05)。结论哮喘疾病中OPN和VEGF含量明显升高,通过减少哮喘小鼠肺组织中OPN和VEGF的含量是细辛脑联合布地奈德减轻哮喘炎症以及改善哮喘早期气道重塑的机制之一。

Objective To investigate the expression of osteopontin(OPN) and vascular endothelial growth factor(VEGF) in asthmatic mice, and the intervention of combination of asarone plus budesonide group on airway remodeling, the levels of OPN and VEGF. Methods BALB/c mice were randomly divided into four groups: control group, asthma group, budesonide group and asarone plus budesonide group, 10 in each group. Modle was established by OVA induction. HE staining was used to observe the pathological changes of lung tissue in mice. The perimeter of bronchial lumen(Pbm), the wall area of bronchial tube(Wat), the wall area of bronchial smooth muscle(WAm) and the number of bronchial smooth muscle cells were measured by using medical image analysis software and standardizing by Pbm; The content of VEGF and OPN in lavage fluid bronchoalveolar(BALF) was detected by enzyme linked immunosorbent assay(ELISA); The content of OPN and VEGF in lung tissue of mice was analyzed by Western blot. The data were analyzed by one-way ANOVA, and the LSD-t method was used to compare the differences between two groups, P<0.05 was statistically significant. Results The level of OPN and VEGF in BALF of asthma group(389.1±10.9, 62.3±10.3) was higher than those in budesonide group(110.4±8.6, 38.1±7.2) and asarone plus budesonide group(97.6±7.9, 30.1±6.8), and the difference had statistically significant(P<0.05, P<0.05). Western blot results showed that the positive expression of OPN and VEGF in asarone plus budesonide group were lower than those in asthma group and budesonide group(P<0.05, P<0.05). Image analysis showed that: WAt/Pbm, WAm/Pbm in asthma group were respectively higher than those in budesonide group(P<0.05, P<0.05); WAt/Pbm, WAm/Pbm and N/Pbm in the asarone plus budesonide group lower than those in the budesonide group(P<0.05, P<0.05, P<0.05). Conclusions The content of OPN and VEGF were significantly increased in asthma, one of the mechanisms that the combination of asarone plus budesonide group alleviate inflammation and improve early airway remodeling in asthma is decreasing the content of OPN and VEGF in the lung tissue.