HGF质粒尾静脉注射促使体内高表达

High expression of HGF in vivo promoted by HGF plasmid injection through the tail vein rapidly

目的:采用质粒尾静脉快速输注以获得肝细胞生长因子(HGF)小鼠体内高表达。方法:扩增HGF质粒,并酶切鉴定。将不同剂量的HGF质粒通过尾静脉快速注入6～8周龄的非肥胖糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠体内,ELISA法检测外周血中HGF水平,同时Western法检测小鼠肝组织内HGF蛋白含量。结果:快速注入HGF质粒后,体内HGF水平明显上升,以20"g/1.6 ml剂量组最高。注射后4 h即可见到HGF水平明显上升,12 h达到顶峰,然后逐渐下降,但6天后仍可测到较高水平的HGF。Western结果显示注射后肝组织中HGF表达量迅速上升,峰值在第8 h左右出现,而后逐步下降。结论:HGF质粒尾静脉快速注射可使小鼠体内HGF高表达。

Objective ：To obtain high expression level of HGF protein in vivo by HGF plasmid rapid injection through the tail vein. Methods ：Systemic administration of different amount of naked plasmid containing HGF eDNA driven under cytomegalovirus promoter （pCMV-HGF） was injected rapidly into NOD/SCID mice through the tail vein, and the level of HGF protein in the circulation was detected by ELISA. The amount of HGF protein in liver tissue was measured through Western blot. Results：The amount of human HGF protein in the circulation was drastically increased after the injection of the HGF vector, and 20 μg/1.6 ml group increased highest. Four hours after the injection of the pCMV-HGF vector, the HGF protein in the circulation of the mice was increased remarkably. Twelve hours later,the concentration of HGF protein reached the maximum that is about 10 ng/ml. Even 6 days after the injection,the HGF protein was still in a high level. The result of Western blot showed that the amount of HGF in liver tissue also increased after injection and reach the peak value at 8 h, and then decreased. Conclusion：Systemic administration of pCMV-HGF via rapid injection through the tail vein can promote high expression of HGF in vivo.