Inhibition of liver fibrosis in vitro using vitamin A-coupled liposomes to deliver MMP-2 siRNA

• 1、Medical school,Southeast University, Nanjing 210009
• 2、Medical school, Southeast University, Nanjing 210009
• 3、Medical school, Southeast University, Nanjing 210009
• 4、School of public health, Southeast University, Nanjing 210009

Abstract：Objective To study the therapeutic potential for liver fibrosis in vitro using vitamin A-coupled liposomes to deliver siRNA against matrix metalloproteinase-2(MMP-2) to hepatic stellate cells(HSCs). Methods The VitA-phosphatidylethanolamine (VitA-PE) compounds, cationic liposomes and vitamin A-coupled liposomes were prepared by DCC-DAMP condensation, rotatory film and ultrasonic oscillation, respectively. MTT assay was performed to investigate the cytoxicity in vitro. HSCs were treated with VitA-lip-MMP-2 siRNA for different hours. RT-PCR and gelatin zymography were utilized to detect the changes of MMP-2 expression and activity. Immunocytochemistry was used to measure the expression of α-SMA andⅠtype collagens in HSCs. Results Before modification, the average diameter of VitA-Lips was 148.2±0.3 nm, besides the average diameter was 227.3±4.1nm. MTT assay showed that VitA-Lips had low cytoxicity. It had good binding ability with siRNA, and could deliver foreign siRNA to HSCs. Treated with VitA-lip-MMP-2 siRNA, the peak of gene silence efficiency in HSCs was 61.84%, MMP-2 mRNA expression and its activity, α-SMA andⅠtype collagens protein expression were significant decline and the proliferation of the cells came slower. Conclusions VitA-lip-MMP-2 siRNA treatment could reduce the proliferation and activation of HSCs and type Ⅰ collagen expression. It suggests that suppress the MMP-2 expression in HSCs may contribute to hepatic fibrosis reverse.

Keywords： liver fibrosis cationic liposomes vitamin A matrix metalloproteinase-2(MMP-2) hepatic stellate cells(HSCs) Small interference RNA(siRNA)