中文摘要：

目的：研究采用气管内注射给予Dectin-1重组腺病毒的方法建立气道Dectin-1上调表达小鼠模型,并进行模型验证和安全性评价.方法：BALB/c小鼠分为PBS组、Ad-EGFP（recombinant adenovirus encoding EGFR）组、Ad-m Dectin-1（recombinant adenovirus encoding murine Dectin-1）组,经气管给予PBS、Ad-EGFP、Ad-m Dectin-1.通过real-time PCR、免疫组化等方法动态评估肺组织Dectin-1表达变化.通过ELISA检测肺组织匀浆及血液中IL-6水平,激光共聚焦检测腺病毒载体在肺组织及肝脏的分布.结果：气管内给药后,在第3天可以观察到Ad-mDectin-1组小鼠肺组织Dectin-1 mRNA水平约为PBS组、Ad-EGFP组的18倍.Ad-m Dectin-1组与对照组比较,差异具有统计学意义（P〈0.05）.给药后第3、5、7、14天,Ad-m Dectin-1组免疫组化见Dectin-1主要定位于细支气管.随时间推移,Dectin-1水平逐渐增高,第5天达高峰,第14天气道上皮细胞仍有Dectin-1表达.给药后第3天,激光共聚焦显微镜观察Ad-mDectin-1显示在肺组织中Ad-mDectin-1组及Ad-EGFP组可见绿色荧光,而PBS组未见绿色荧光.在肝脏组织中,各组均无绿色荧光蛋白表达.肺组织及血清中IL-6水平检测示各组差异无统计学意义（P〉0.05）.结论：通过气管内给予Ad-mDectin-1成功上调小鼠肺组织Dectin-1水平,Dectin-1主要表达于细支气管上皮细胞,表达高峰在第5天,可持续2周以上.另外,经气管给予Ad-mDectin-1对小鼠生存和活动无明显影响,无明显炎症反应及远隔器官肝脏的播散.

英文摘要：

AIM： To constructe a murine model of upregulation of Dectin-1 in the airway by intratracheal administration of Ad-m Dectin-1 and study on identification and security of the murine model. METHODS： BALB / c mice were divided into PBS group,Ad-EGFP group and Ad-m Dectin-1 group,intratracheal administrated with PBS,Ad-EGFP and Ad-m Dectin-1 respectively. After administration,the levels of Dectin-1 mRNA in lung tissue were tested by real-time PCR and immunohistochemitry,the levels of IL-6 in lung homogenate and serum were detected by ELISA,distribution of adenovirus vector in lung and liver was detected by confocal. RESULTS： At day 3 after administration of Ad-m Dectin-1,the Dectin-1 mRNA expression was obviously increase in mice treated with Ad-m Dectin-1,and was resulted in nearly eighteen fold increase compared with mice treated by Ad-EGFP or PBS. For mice treated with Ad-m Dectin-1,immunohistochemistry results showed strong immunostaining in the airways epithelium in bronchioles and nearly negative immunostaining in alveolar walls,the immunostaining of Dectin-1 can last for two weeks. The positive particles were mainly located in cytoplasm and membrane of epithelial cells. In contrast,mice treated with PBS or Ad-EGFP did not detected immunostaining in the lungs. Immunostaining results showed time-dependent change of average optical density with peak reach at day 5 after administration. We used confocal microscopy to detect the green fluorescent protein which can emit green fluorescence under excitation of 488 nm laser. We found green fluorescence in lung but not in liver. Results of ELISA showed no significantly differences of levels of IL-6 were founded compared with mice treated with Ad-EGFP or PBS. Pathology of lung tissue of mice treated with Ad- m Dectin-1 showed mild inflammation reaction,characterized by polymorphonuclear cells infiltration at 3 day post-infection and peribronchial lymphocytes hyperplasia after 3 days. Necrosis and structure changes were not found. CONCLUSION： After intratracheal ad