摘要
目的 为利用热休克蛋白 70 (heat shockprotein,HSP70 )防治严重烧伤后缺血缺氧性损伤的基因治疗提供可行的基因转染途径。 方法 利用微胶囊技术包裹含HSP70基因的复制缺陷型重组腺病毒载体 ,检测其在人工胃、肠液中的融出率 ,并进行大鼠的口服转染 ,检测目的基因表达。 结果 用微胶囊技术可成功包裹腺病毒载体。制备的微胶囊在人工胃液中仅有少量崩解 ,而在肠液中则半数崩解。RT PCR检测大鼠口服转染微胶囊后目的基因表达确切。 结论 利用微胶囊技术可成功包裹含HSP70基因的腺病毒载体 ,并保护其免受胃液损伤 ,在肠道中被释放吸收。
Objective To find a new and effective way for the transfection of adenovirus vectors encoding HSP70 cDNA, so as to provide another possible method in gene therapy against ischemia and cellular hypoxia after burn injury. Methods The replicated defective adenovirus vectors encloding HSP70 cDNA were encapsulated. Its acid resistance and dissolution in intestinal fluid were tested in artificial gastric juice and intesitinal fluid. The expression of HSP70 gene which was transfected by the microcapsules orally was detected by RT-PCR. Results The encapsulated replicated defective adenovirus vectors were viable in vitro. They exhibited good resistant to acid (resolution ratio less than 10%) and dissolution in intestinal juice(resolution ratio higher than 50%). The HSP70 gene expression of the tested rats was significantly higher than control, but there was no difference in the quantity of HSP70 induced by sodium arsenite or adenovirus transfection through injection by vein. Conclusion The encapsulation of adenovirus vectors can successfully keep the viability of the virus in vitro and protect the virus from harmful effect of acid and enzyme in the gastric juice. Its nice dissolution in intestinal juice should ensure its absorption by oral transfection. The expression of the HSP70 gene after oral intake of this preparation is as high as that with other traditional transfection methods. It is possible that in the future the encapsulated replication of defective adenovirus vectors encoding HSP70 cDNA can provide a safer, convenient and effective way for gene therapy for burn patients.
出处
《中华烧伤杂志》
CAS
CSCD
2003年第3期145-147,共3页
Chinese Journal of Burns
基金
国家重点基础研究发展规划资助项目 (G19990 5 42 0 2 )
国家杰出青年科学基金资助项目 (3 0 12 5 0 40 )
