重组腺病毒介导LacZ基因在大鼠肺脏的表达  

Expression of recombinant adenovirus-mediated LacZ gene in rat lung

作　　者：金强[1] 陈卫民[1] 张铁铮[2] 王凤学[2] 周锦[2] 尤圣武[3] 刘晓江[2] 

机构地区：[1]中国医科大学附属盛京医院麻醉科,沈阳市110004 [2]沈阳军区总医院麻醉科 [3]上海交通大学医学院瑞金医院麻醉科

出　　处：《中华麻醉学杂志》2006年第9期856-859,共4页Chinese Journal of Anesthesiology

摘　　要：目的探讨重组腺病毒介导的β-半乳糖苷酶（LacZ）基因在大鼠肺脏的转基因表达。方法Wistar大鼠70只，随机分为Ad—Null组和Ad-LacZ组（n=35），分别应用1．67×10^9pfu/ml复制缺陷型重组腺病毒AdCMV和AdCMVLacZ各600μl，经气管导管滴入；各组于滴入病毒后2、5、7、14、21、28和35d行肺组织X-gal染色。另取大鼠20只，随机分为C组、L组、M组和H组（n=5），分别应用病毒保存液和低滴度（1．67×10^8pfu／ml）、中滴度（1．67×10^9pfu／ml）、高滴度（5×10^9pfu／ml）的AdCMVLacZ各600μl，滴入病毒后7d行肺组织X-gal染色和HE染色。结果滴入病毒后2d，Ad-LacZ组肺组织内即有转基因表达，滴入后7d达高峰，并维持至35d；转基因表达位于气管、支气管上皮细胞和肺泡细胞。H组、M组转基因阳性细胞率明显高于L组和C组（P〈0．01）。HE染色显示，H组中3只大鼠肺组织有轻度炎性浸润。所有动物均未见远隔器官转基因表达。结论气管内滴入重组腺病毒可呈剂量依赖性介导LacZ基因在大鼠肺内表达，但过高剂量可诱发机体炎性反应。Objective To investigate the expression of recombinant adenovirus-mediated LacZ gene in rat lung after intratracheal instillation. Methods Seventy male Wistar rats weighing 220-280 g were randomly divided into 2 groups： Ⅰ Ad-LacZ group （ n = 35） and Ⅱ Ad-null group （ n = 35）. The recombinant adenovirus Ad-LacZ was produced in Ad 293 cells. The rats received 600 μl of AdCMV LacZ or AdCMV （1.67 × 10^9 pfu·ml^-1 ） by intratracheal instillation. The animals were killed at 2, 7, 14, 21, 28 and 35 d after intracheal adenovirus administration, and lung specimen were examined by X-gal staining. Another 10 rats were randomly divided into 4 groups （ n = 5 each） ： group Ⅰ high dose; group Ⅱ medium dose, group Ⅲ low dose and group Ⅳ control. The rats were given AdCMV LacZ 5 × 10^9（ Ⅱ ）, 1.67 × 10^9（Ⅱ ）, 1.67 ×10^8（Ⅲ） pfu·ml^-1 or vector conservation solution （ Ⅳ ） 600 μl by intratracheal instillation. The animals were killed at 7 d after intratracheal instillation and X-gal and HE staining of the lung specimens were performed. Results The LacZ gene expression was found in the lung at 2 d after AdCMV LacZ administration, and peaked at 7 d and was sustained for 5 weeks. LacZ expression was detected in the epithelial cells of trachea, bronchi, bronchioles and alveolar cells and was significantly higher in high and medium dose groups than in low dose and control groups. Mild inflammatory infiltration of the lung was foumd only in 3 out of 5 animals in high dose group. No LacZ gene expression was found in distant organs. Conclusion Recombinant adenovirus-mediated LacZ gene can be transferred in the lung efficiently by intratracheal instillation in a dose-dependent manner. High dose of vector may induce immune response.

关 键 词：β半乳糖苷酶类 腺病毒科 基因转移技术 肺 

分 类 号：R614[医药卫生—麻醉学] R563[医药卫生—外科学]