Collagen-binding vascular endothelial growth factor attenuates CCl4-induced liver fibrosis in mice.

ABSTRACT

Vascular endothelial growth factor (VEGF) serves an important role in promoting angiogenesis and tissue regeneration. However, the lack of an effective delivery system that can target this growth factor to the injured site reduces its therapeutic efficacy. Therefore, in the current study, collagen­binding VEGF was constructed by fusing a collagen­binding domain (CBD) to the N­terminal of native VEGF. The CBD­VEGF can specifically bind to collagen which is the major component of the extracellular matrix in fibrotic liver. The anti­fibrotic effects of this novel material were investigated by the carbon tetrachloride (CCl4)­induced liver fibrotic mouse model. Mice were injected with CCl4 intraperitoneally to induce liver fibrosis. CBD­VEGF was injected directly into the liver tissue of mice. The liver tissues were stained with hematoxylin and eosin for general observation or with Masson's trichrome staining for detection of collagen deposition. The hepatic stellate cell activation, blood vessel formation and hepatocyte proliferation were measured by immunohistochemical staining for α­smooth muscle actin, CD31 and Ki67 in the liver tissue. The fluorescent TUNEL assay was performed to evaluate the hepatocyte apoptosis. The present study identified that the CBD­VEGF injection could significantly promote vascularization of the liver tissue of fibrotic mice and attenuate liver fibrosis. Furthermore, hepatocyte apoptosis and hepatic stellate cell activation were attenuated by CBD­VEGF treatment. CBD­VEGF treatment could additionally promote hepatocyte regeneration in the liver tissue of fibrotic mice. Thus, it was suggested that CBD­VEGF may be used as a novel therapeutic intervention for liver fibrosis.