摘要目的 评价腺病毒自身的致炎作用在以其为载体的基因治疗中,对小鼠急性肺损伤(ALI)治疗效果的影响及意义.方法 C57/B6雄性小鼠随机分为4组:(1)以腺病毒为载体的靶基因LacZ治疗组(AdLacZ组):43只;(2)3个对照组:①O2吸入前组(Con组):26只;②O2吸人后组:TBS+磷酸盐缓冲液(PBS)治疗组(PBS组)36只;③无治疗对照组(无治疗组):33只.100%O2吸入诱导小鼠高氧性ALI动物模型;在100%O2吸入前48 h经鼻腔通道给予装载LaeZ DNA的腺病毒质粒感染小鼠,并观察小鼠的死亡率.B-半乳糖酶活性测定法用于确定转染的1acZ基因在肺脏的蛋白活性的表达水平;肺的干/湿重比、支气管-肺泡灌洗液内的蛋白浓度和细胞分类分析用于评价肺炎症反应的程度.结果 经鼻腔通路的以腺病毒为载体的基因治疗,可造成转染基因LacZ的蛋白表达活性在肺脏持续有效地高表达,而且其表达水平在O2吸人72 h后仍可维持为O:吸入前的2倍以上(3.688 U/mg v8 1.589 U/mg);该组小鼠对高氧吸入敏感性高于各对照组,50%的小鼠生存时间短于PBS组[(86±3)h vs(94±7)h];同时,该组小鼠的支气管-肺泡内炎症细胞的渗出在100%O2吸入48 h后明显高于各对照组,继续O2吸入24 h后明显下降约50%;肺干/湿重比和支气管-肺泡灌洗液内蛋白浓度无明显下降.结论 腺病毒自身的致炎作用对ALI基因治疗效果的影响是轻度和暂时的,但需在评价治疗效果时加以重视.

abstractsObjective To evaluate the impact of endogenous inflammation caused by adenovims as a vector of gene therapy on monse model of acute lung injury(ALI).Methods black C57/B6 mice randomly divided into 4 research groups:(1)adenovirus encoded-lacZ genetreatment group(AdLacZ):n=AdLscZ reagent through intranasal administration to infect mice lungs at 48h before 100%O2 inhalation to induce ALI mouse model,which companied by mice survival rates recorded.B-gal protein activity in lung wag detected to show the level of LacZ DNA transgenic protein activity;meanwhile,the indices of lung wet/dry ratio and bronchial alveolar lavage liquid(BALF)analysis with protein concentration and cell elagsification were detected.Results The method of adenovirus-mediated gene therapy with intranagal administration restthed in IJacZ DNA transgenic protein activity to keep effective highly expression in lung.and the expression level maintained 2.fold increusing even after 72 h of O.inhalation compared to that before O2 inhalation(3.688 U/mg vs 1.589 U/mg);AdLacZ mice had more subjective to O2 inhalation compared to other groups.the 50%mice survival time of this group wag shorter compared to that of the PBS group [(86±3)h vs(94±7)h];also in AdLacZ group,the level of nucleated cell counting in BALF was statistically higher compared to other groups at 48 h of O2 inhalation,which following with 50%-level decreased within anther 24 h O2 inhalation;on the contrary,the level of lung wet/dry ratio and protein concentration in BALF didn,t show remarkably decreasing.Conclusion The endogenous inflammation caused by adenovirus as a vector in ALl gene therapy is temporary and rapidly weaken after getting a peak,however,enough attention still should be paid attention when evaluating the effect of gene therapy.