Purification of Antiangiogenesis Compound from Indonesian Shark Cartilage (Elasmobranchii) and In Vitro Assay with Endothelial Cell Culture

Abstract

Shark cartilage has been recognized as an alternative medicine for cancer and other diseases caused by the angiogenesis process. In this study, the antiangiogenesis effect of crude extract, purified glycoprotein and polysaccharide of shark cartilage were analyzed at various concentrations (400, 600 and 800 ppm). Antiangiogenesis assay was measured by analyzing the proliferation inhibition of capillary pulmonary artery endothelium cell culture (CPAE-ATCC CCL-209). Chromatography gel filtration with Sephadex G-75 was used to extract the glycoprotein compound. The process yielded 17.4% as crude glycoprotein. Further purification was performed with DEAE Sephadex A-25. Fractionation based on ultraviolet absorption at 280 nm and qualitative assay resulted into four fraction and yielded 0.009% - 0.32% of fraction. Extraction and fractionation of polysaccharide were performed by gel filtration with Toyopearl HW-65F. Three fractions were collected and yielded 0.61% - 2.26% of fraction. The result of in vitro antiangiogenesis assay showed that crude extract and fractions of glycoprotein and potysaccharide gave an inhibition on the proliferation of endothelial cell culture. The crude extract of glycoprotein (800 ppm) strongly showed 100% proliferation inhibition. From the po\ysaccharide fraction, fraction Ill that contain the highest concentration of polysaccharide and the lowest concentration of protein showed 98.9% - 10O0/0 of inhibition. This study indicates that purified compound from lndonesian shark cartilage has potential in vitm antiang iogenesis effect.