An experimental study of a modified Dahuang Zhechong pill on the-angiogenesis of RF/6A cells in vitro

Abstract

ObjectiveTo investigate the effects of a modified Dahuang Zhechong Pill (MDZP) on the angiogenesis of rhesus choroid-retina endothelial (RF/6A) cells and its preliminary mechanism. MethodsA 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method was used to assess the effect of a MDZP on RF/6A cell proliferation induced by vascular endothelial growth factor (VEGF). Transwell inserts were used to assess the effect of the MDZP on RF/6A cell migration. Matrigel was used to assess the effect of the MDZP on the tube formation of RF/6A cells. Western blotting and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression, respectively, of VEGF and matrix metalloproteinase-2 (MMP-2) in RF/6A cells treated with the MDZP. ResultsRF/6A cell proliferation induced by VEGF was inhibited by 0.2 mg/mL MDZP. At 0, 12.5, 25 and 50 mg/mL MDZP, the number of cells that migrated through Transwell membranes was 73.33 ± 4.51, 61.33 ± 4.04, 28.67 ± 6.66 and 17.67 ± 4.16, respectively, and the number of tubes formed in Matrigel was 20.33 ± 0.58, 13.33 ± 1.53, 11.00 ± 1.00 and 1.33±0.58, respectively. At 100 and 200 mg/mL MDZP, the protein and mRNA expression of VEGF and MMP-2 were inhibited in RF/6A cells. At 400 mg/mL MDZP, the expression of VEGF mRNA and MMP-2 protein were inhibited in RF/6A cells. ConclusionsMDZP inhibits the angiogenesis of RF/6A cells via the suppression of proliferation, migration and tube formation of RF/6A cells. Inhibition of the protein and mRNA expression of VEGF and MMP-2 in RF/6A cells may be an important mechanism.