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

来源 :Journal of Traditional Chinese Medicine | 被引量 : 0次 | 上传用户:yyl273518021
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OBJECTIVE:To investigate the effects of a modified Dahuang Zhechong Pill(MDZP) on the angiogene sis of rhesus choroid-retina endothelial(RF/6A cells and its preliminary mechanism.METHODS:A 3-(4,5-dimethylthiazol-2-yl)-5-(3-car boxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazol ium(MTS) method was used to assess the effect o a MDZP on RF/6A cell proliferation induced by vas cular endothelial growth factor(VEGF).Transwell in serts were used to assess the effect of the MDZP on RF/6A cell migration.Matrigel was used to asses the effect of the MDZP on the tube formation of RF 6A cells.Western blotting and quantitative re al-time reverse transcription polymerase chain reac tion(RT-PCR) were used to detect the protein and mRNA expression,respectively,of VEGF and matri metalloproteinase-2(MMP-2) in RF/6A cells treatedwith 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. OBJECTIVE: To investigate the effects of a modified Dahuang Zhechong Pill (MDZP) on the angiogene sis of rhesus choroid-retina endothelial (RF / 6A cells and its preliminary mechanism. METHODS: A 3- (4,5-dimethylthiazol- ) 5- (3-car boxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (MTS) method was used to assess the effect oa MDZP on RF / 6A cell proliferation induced by vas cular endothelial growth factor ). Transwell in serts were used to assess the effect of the MDZP on RF / 6A cell migration. Matrigel was used to asses the effect of the MDZP on the tube formation of RF 6A cells. Western blotting and quantitative re al-time reverse transcription respectively, of VEGF and matri metalloproteinase-2 (MMP-2) in RF / 6A cells treated with the MDZP.RESULTS: RF / 6A cell proliferation induced by polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression, 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 mem branes 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 important mechanism .
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