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

作者：Luo Xu-sheng; Wu Xing-wei^*; Gu Qing

来源：Journal of Traditional Chinese Medicine, 2012, 32(1): 75-81.

DOI：10.1016/S0254-6272(12)60036-3

• Modified dahuang zhechong pill
• Rhesus choroid-retina endothelian cell line
• Proliferation
• Migration
• Tube Formation
• Angiogenesis
• Choroidal neovascularization
• Vascular endothelial growth factor
• matrix metalloproteinase-2

OBJECTIVE: To 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. METHODS: A 3-(4, 5-dimethylthiazol-2-y1)-5-(3-carboxymethoxypheny1)-2-(4-sulfopheny1)-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.
RESULTS: RF/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.
CONCLUSIONS: MDZP 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.

• 出版日期2012-3
• 单位中国中医科学院眼科医院; 上海交通大学