LacZ DNA标记的腺病毒在高氧性肺损伤基因治疗中的转染和致炎作用  

作　　者：王燕 AronB．Fisher FeisteinSSheldon 张灵恩[3] 

机构地区：[1]上海市徐汇区中心医小儿科 [2]the Institute of EnvironmentalMedicine,the Center of Lung Biology,University of Pennsylvania,USA [3]上海复旦大学医学院儿科医院ICU

出　　处：《新课程学习（下）》2012年第2期10-12,共3页

摘　　要：目的：LacZDNA标记的腺病毒在高氧性肺损伤基因治疗中的转染和致炎作用。方法：装载LacZDNA的腺病毒在小鼠高氧模型开始前2天经鼻腔送入体内X-lal染色和β一半乳糖酶活性测定用于确定LacZDNA在肺内转染的分布及其蛋白表达产物的活性；小鼠肺干，湿重比和支气管肺泡盥洗液（BALF）内的蛋白浓度的测定用于鉴定腺病毒在基因治疗过程中可能存在的致炎作用。结果：X-gal染色显示LacZDNA在高氧前及后均可广泛转染在肺各级支气管和肺泡上皮细胞，β一半乳糖酶活性的测定提示了LacZDNA可成功地翻译成其蛋白质表达产物并表现为高表达；同时，腺病毒的致炎作用在高氧吸入48h后示小鼠肺干，湿重比和BALF蛋白浓度均较其他对照组明显升高，但其升高的程度可在24h后迅速缓解上升趋势。结论：腺病毒为载体的基因治疗可有效地转染标记基因，同时，腺病毒虽在基因治疗的过程中有一定的致炎作用，但是暂时的，其仍是有效的DNA载体。Objective：To investigate the effects of gene transfer and endogenous inflammation of adenovirus marked by LacZ DNA to hyperoxia lung injury in gene therapy.Methods：Adenovirus encoded LacZ DNA was intranasal administered to mice at 2 days earlier before 100%O2 inhalation, X--gal staining and activity measurement of [3-gal protein translated from LacZ DNA were applied to determine the level of LacZ DNA transfer and it＇s protein expression; meanwhile, mouse lung wet/dry ratio and protein concentration in b^onchus alveolar fluid were measured to valuate the degree of inflanunation reaction from adenovirus.Results：X-gal staining showed LacZ DNA could transfer broadly in series of lung bronchus and alveolar epithelial cells no matter before or after 100%02 inhalation; 13-gal protein activity measurement presented LaeZ DNA could successfully translated into it＇s protein expression with high level expression＇meanwhile, endogenous inflammation of adenovirus showed moose lung wet/dry ratio and protein concentration in bronchus alveolar fluid were significantly increased at the 48h of 100%02 inhalation compared to control groups, but lost the pattern of continue increasing after anther continual 24 h of 100%02 inhalation.Conclusion：being vector of gene therapy, adenovirus could successfully transfer target DNA, while, adenovirns would bring some degree of endogenous inflammation, but it is tealporarily, it is still a validity vector in gene therapy.

关 键 词：腺病毒 基因治疗 LACZ 高氧性肺损伤 载体 

分 类 号：Q78[生物学—分子生物学]