摘要
目的 将动脉插管方法应用于重组腺病毒 (Ad .LacZ)介导的标记基因体内导入 ,观察标记基因在大鼠内脏中的转染效率和维持时间。方法 9组大鼠 ,7组于胃十二指肠动脉分别注入Ad .LacZ[2× 10 9pfu/ml(空斑形成单位 /ml) ]各 1ml,2组注入单纯培养液各 1ml,7组实验组分别于 12h、18h、72h、7d、14d、2 1d、2 8d取动物的肝、脾、肺和肾脏 ,2组对照组于 7d取上述脏器 ,采用 β 半乳糖苷酶 (X Gal)染色方法检测LacZ基因在各个脏器的表达。结果 实验组于 12h取材 ,经染色LacZ基因在肝脏中即有表达 ,而此时脾、肺、肾等内脏器官尚未见表达 ;此后 ,18h、72h、7d、14d直至 2 1d该基因在肝脏仍有表达 ,而在其他脏器如脾、肺及肾表达仅维持 14d。对照组动物各个脏器皆无染色。结论 经动脉途径注入重组腺病毒 ,其体内的表达具有器官靶向性。如经此途径注入治疗性的目的基因重组腺病毒 ,其于肝脏表达产物出现时间早 ,并且持续时间较长。
Objective To investigate the gene transfer efficiency and lasted time in rat organs by hepatic artery injection with LacZ reporter gene recombinant adenoviruses. Methods Seven groups of rats were injected with Ad.LacZ (2×10 9 pfu/ml) and two groups of rats were injected with PBS 1 ml as control separately through gastra intestinal artery, and liver, spleen, lung, and kidney were gotten at 12 hrs, 18 hrs, 72 hrs, 7 day, 14 day, 21 day, and 28day, respectively. X gal staining was used to check up expression of LacZ gene. Results Expression of LacZ gene was detected in liver 12 hrs after injection, but none were done in spleen, lung, and kidney. Up to 21days, LacZ gene expressed in liver, but the gene expression lasted for only 14 days in spleen, lung, and kidney LacZ gene was not detected in the two control groups in all organs at 7 day. Conclusion When recombinant adenovirus was administrated through hepatic artery, the introduced gene expressed preferentially in liver. This result was the basis of intraarterial administration of cytokines gene to treat liver tumor.
出处
《中华放射学杂志》
CAS
CSCD
北大核心
2000年第10期662-665,共4页
Chinese Journal of Radiology
基金
国家"九五"科技攻关计划资助项目!(96 90 6 0 1 2 0 )
关键词
肿瘤
基因治疗
重组腺病毒
基因转染
肝动脉插管
Animal testing alternatives
Radiology
interventional
Catheterization
Gene transfer
Transfection
Adenoviridae
Biological markers
Gene expressiX
