摘要
目的建立大鼠体内质粒DNA尾静脉快速大容量注射法,使外源性rIL-10在肝组织中高表达,为探索肝脏疾病的基因治疗提供动物试验手段。方法以rIL-10为报告基因,将含不同体积及不同质粒浓度的质粒DNA溶液经大鼠尾静脉快速注射,在注射后的不同时间内分别收集血浆及肝、肾、肺、脾和心组织,使用RT-PCR、ELISA和免疫组化法检测rIL-10在体内的表达情况。结果尾静脉快速大容量注射rIL-10DNA溶液可使rIL-10基因在大鼠肝脏有较明显转录及表达。血浆中rIL-10浓度可通过质粒的重复注射使其保持在一个稳态。结论尾静脉快速大容量注射法是一种简单、方便和有效的大鼠体内基因转移及基因表达的方法,为进一步探讨rIL-10基因在肝脏疾病的基因治疗打下基础。
Ahn To establish a method of hydrodynamic-based transfection （HBT） and provide a means in rats of the gene therapy of liver diseases,which allowed an efficient expression of rlL-10 gene in rat liver. Methods Using rlL-10 gene as a reporter gene, different volumes and doses of plasmid DNA solutions were rapidly injected into rat tail vein, then the serum and the tissue of liver,kidney ,lung,spleen and heart in different time were collected and the expression of rlL-10 was detected by the methods of RT-PCR, ELISA and immunochemistry. Results Using rIL-10 gene as a reporter gene,the results demonstrated that an efficient transfer and expression of rlL-10 in rat liver could be achieved by a rapid injection of a large volume of rlL-10 DNA solution into rat via tail vein. Maintaining a stable expression of rlL-10 in serum could be assessed by repeated administration. Conclusion The HBT was a simple, convenient and efficient method of gene transfer and expression in rats, which could be used as an effective means to study further gene therapy of rIL- l0 in liver diseases.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第6期834-837,共4页
Chinese Pharmacological Bulletin
基金
福建省科技开发计划资助项目(No2005D094)
