新课程学习：下
新課程學習：下
신과정학습：하
2012年
2期
10-12
,共3页
王燕%AronB．Fisher%FeisteinSSheldon%张灵恩
王燕%AronB．Fisher%FeisteinSSheldon%張靈恩
왕연%AronB．Fisher%FeisteinSSheldon%장령은
腺病毒%基因治疗%LacZ%高氧性肺损伤%载体
腺病毒%基因治療%LacZ%高氧性肺損傷%載體
선병독%기인치료%LacZ%고양성폐손상%재체
adenovirus%gene therapy%LacZ%hyperoxia hmg injury%vector
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。
目的：LacZDNA標記的腺病毒在高氧性肺損傷基因治療中的轉染和緻炎作用。方法：裝載LacZDNA的腺病毒在小鼠高氧模型開始前2天經鼻腔送入體內X-lal染色和β一半乳糖酶活性測定用于確定LacZDNA在肺內轉染的分佈及其蛋白錶達產物的活性；小鼠肺榦，濕重比和支氣管肺泡盥洗液（BALF）內的蛋白濃度的測定用于鑒定腺病毒在基因治療過程中可能存在的緻炎作用。結果：X-gal染色顯示LacZDNA在高氧前及後均可廣汎轉染在肺各級支氣管和肺泡上皮細胞，β一半乳糖酶活性的測定提示瞭LacZDNA可成功地翻譯成其蛋白質錶達產物併錶現為高錶達；同時，腺病毒的緻炎作用在高氧吸入48h後示小鼠肺榦，濕重比和BALF蛋白濃度均較其他對照組明顯升高，但其升高的程度可在24h後迅速緩解上升趨勢。結論：腺病毒為載體的基因治療可有效地轉染標記基因，同時，腺病毒雖在基因治療的過程中有一定的緻炎作用，但是暫時的，其仍是有效的DNA載體。
목적：LacZDNA표기적선병독재고양성폐손상기인치료중적전염화치염작용。방법：장재LacZDNA적선병독재소서고양모형개시전2천경비강송입체내X-lal염색화β일반유당매활성측정용우학정LacZDNA재폐내전염적분포급기단백표체산물적활성；소서폐간，습중비화지기관폐포관세액（BALF）내적단백농도적측정용우감정선병독재기인치료과정중가능존재적치염작용。결과：X-gal염색현시LacZDNA재고양전급후균가엄범전염재폐각급지기관화폐포상피세포，β일반유당매활성적측정제시료LacZDNA가성공지번역성기단백질표체산물병표현위고표체；동시，선병독적치염작용재고양흡입48h후시소서폐간，습중비화BALF단백농도균교기타대조조명현승고，단기승고적정도가재24h후신속완해상승추세。결론：선병독위재체적기인치료가유효지전염표기기인，동시，선병독수재기인치료적과정중유일정적치염작용，단시잠시적，기잉시유효적DNA재체。
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.
