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

Abstract

Hepatic fibrosis is a common form of wound healing in response to chronic liver injuries and can lead to more serious complications, including mortality. It is well-established that hepatic stellate cells (HSCs) are central mediators of hepatic fibrosis, and matrix metalloproteinase-2 (MMP-2) is important in the formation of liver fibrosis. In addition, HSCs are the primary cells secreting MMP-2 and extracellular matrix, therefore, there has been increasing interest in developing agents with high selectivity towards HSCs. However, no clinical drugs based on MMP-2, directed against HSCs, have been used to prevent fibrosis. Following consideration of the abundant vitamin A (VitA) receptors expressed on the cellular membrane of HSCs, the present study constructed VitA-coupled liposomes (VitA-lips) using dicyclohexylcarbodiimide-1, 3-diaminopentane condensation, rotatory film processing and ultrasonic oscillation. The results revealed that the liposomes exhibited low cytotoxicity and a suitable binding ability to MMP-2 small interference (si)RNA. Furthermore, the liposomes effectively delivered MMP-2 siRNA to the HSC-T6 cells. When HSCs were treated with the liposomes carrying MMP-2 siRNA (VitA-lip-MMP-2 siRNA), the mRNA expression and activity of MMP-2, and the protein expression levels of α-smooth muscle actin and type I collagen were significantly reduced. These results suggested that inhibition of the expression of MMP-2 in HSC-T6 cells may contribute to preventing hepatic fibrosis, and provided experimental support to the development of specific drugs against MMP-2 to prevent fibrogenesis in chronic liver disease.