Effect of exogenous human urokinase-type plasminogen activator gene on the production of collagens in hepatic stellate cells / 中华消化杂志

ABSTRACT

Objective To study the effect of exogenous non secreted human urokinase type plasminogen activator(uPA) gene on the production of collagen in activated hepatic stellate cells(HSC) after delivery by replication deficient recombinant adenovirus vector. Methods Non secreted human uPA was inserted into the replication deficient recombinant adenoviruses AduPA in bacteria. After infected by AduPA, the expression of uPA in HSC cell line HSC T6 was detected by Northern blot and immunocytochemical staining, and the level of matrix metalloproteinase 2 (MMP 2) in the supernatant of cultured HSC T6 cells was measured by ELISA. In addition, immunocytofluorescent staining was carried out to evaluate the effect of AduPA on the production of collagen Ⅰ and Ⅲ in HSC after exogenous uPA gene delivery. Results 5?10 11 efu/ml titer of AduPA was obtained in the HSC infected with the recombinated virus. Northern blot and immunocytochemical staining showed that the expression of uPA in HSC increased significantly on the third day after infection with the AduPA. The level of MMP 2 in the supernatant of the cultured HSC was (274.45?7.63) pg/ml, which was higher than that in the control [(145.85?6.58) pg/ml, P