多聚胺胆固醇阳离子脂质体介导CpGODN雾化吸入对哮喘小鼠肺组织嗜酸性粒细胞的影响

Influence of polyamine-chol cationic liposome-mediated CpGODN aerosol on the eosinophiles in the lung tissue of mouse with asthma

目的探讨雾化吸入多聚胺胆固醇阳离子脂质体(PCL)与CpGODN复合物(CpGODN/PCL复合物)对哮喘小鼠肺组织嗜酸性粒细胞(EOS)的影响。方法采用卵蛋白激发法建立哮喘小鼠模型,并设置正常对照组、哮喘对照组、CpGODN/PCL干预组、CpGODN干预组,每组6只。在雾化PCL介导转染CpGODN 48h后,分离小鼠左肺组织,切片HE染色,显微镜下检测EOS浸润情况,同时检测小鼠支气管肺泡灌洗液(BALF)中细胞浸润情况。结果用卵蛋白激发法成功地制备了哮喘小鼠模型。哮喘对照组肺组织肺泡腔黏液分泌增加,支气管及血管周围大量炎性细胞浸润,以EOS、淋巴细胞为主,与正常对照组比较,BALF中细胞总数、EOS数目和百分率均明显增加(P<0.01)。而与哮喘对照组比较,CpGODN干预组细胞总数,EOS数目和百分率显著下降(P<0.01)。CpGODN/PCL干预组与CpGODN干预组比较,细胞总数无统计学差异,EOS数目、百分率的差异有统计学意义(P<0.05,P<0.01)。结论哮喘小鼠肺组织EOS浸润和气道黏液分泌,雾化介导CpGODN干预哮喘小鼠模型可减轻以EOS为特征的炎症反应,利用PCL为转染载体包裹CpGODN,可提高转染效率。

Objective To explore the influence of polyamine-cholesterol cationic liposome （PCL）-mediated CpGODN aerosol on eosinophiles in the lung tissue of mouse asthma model. Methods Mouse asthma model was replicated by challenging with 1% ovalbumin aerosol. Mice were categonied into four groups, namely normal control, asthma control, CpGODN/PCL treatment group and CpGODN treatment group （6 each）. The left lungs of mice were harvested, serially sectioned, hematoxylin and eosin （HE）-stained, and the infiltration of eosinophiles （EOS） was examined under microscope. Meanwhile, the bronchoalveolar lavage fluid （BALF） was collected for total and eosinophil cells count. Results An ovalbumin challenged mouse asthma model was successfully replicated. Pathological observation of the lung of asthma control showed increase in mucous secretion in alveolar space and peribronchial infiltration of large amount of inflammatory cells, primarily EOS and lymphocytes. The total cell number, EOS number and the ratio in BALF were significantly higher in asthma control group compared with that in both normal control group and CpGODN treatment group （P〈0.01）. No significant difference was found in the total cell number, but the number and ratio of EOS were significantly different （P〈0.05, P〈0.01） compared between CpGODN/PCL treatment group and CpGODN treatment group. Conclusions An ovalbumin-challenged mouse asthma model has been successfully replicated. The present study demonstrates that CpGODN aerosol treatment can abate inflammatoary reactions, characterized by EOS infiltration. CpGODN PCL in capsule, has a better transfection efficacy than the bare-CpGODN.