双氢青蒿素含药血清抗血管生成活性的研究

The Research on The Anti-angiogenic Activity of Dihydroartemisinin Contained Serum

目的:研究不同剂量组双氢青蒿素对人脐静脉内皮细胞的增殖、迁移抑制作用和作用机制。方法:制备空白对照组、双氢青蒿素15 mg/kg组、30 mg/kg组和60 mg/kg组4个剂量组含药血清,以贝伐单抗为对照药作用于人脐静脉内皮细胞,噻唑蓝法检测细胞增殖,划痕实验检测细胞迁移能力,RT-q PCR法检测细胞VEGF表达,放射配体受体结合分析法检测细胞VEGFR最大结合容量和解离常数。结果:相对于空白对照组,双氢青蒿素30 mg/kg组和60 mg/kg组可明显抑制人脐静脉内皮细胞增殖且能降低细胞的迁移能力;RT-q PCR结果显示,这2个剂量组的VEGF表达出现明显降低;VEGFR最大结合容量出现明显降低,解离常数明显升高,结果均具有显著性意义。结论:30 mg/kg剂量以上的双氢青蒿素含药血清可显著抑制人脐静脉内皮细胞增殖和迁移,作用机制可能与抑制血管内皮生长因子表达并阻断其与受体的结合相关。

Objective： To study the proliferation and migration ability impact of dihydroartemisinin of different dosage group on human umbilical vein endothelial cell（HUVEC） and its mechanism.Methods： Prepared four dosage groups of drug serum,i.e.blank control group,dihydroartemisinin 15 mg/kg group,dihydroartemisinin 30 mg/kg group and dihydroartemisinin 60 mg/kg group and then used them to HUVEC.Detected the cell proliferation with MTT method; The scratch test was used to analyze the cell migration ability; detected the VEGF expression with RT-q PCR method and detected VEGF receptors＇ Bmax and dissociation constant with RBA method.Results： Compared with blank control group,the dihydroartemisinin 30 mg/kg group and dihydroartemisinin 60 mg/kg group obviously inhibited cell proliferation and reduce the ability of cell to migrate; RT-q PCR results show that the expression of dihydroartemisinin 30 mg/kg group and dihydroartemisinin 60 mg/kg group has significantly decrease; VEGF receptors Bmax in these two groups decreased clearly while the dissociation constant increased obviously.The results all have statistical significance.Conclusion： More than 30 mg/kg dose of dihydroartemisinin can remarkably inhibit HUVEC proliferation and migration ability.The inhibition mechanism may be related with the inhibition of VEGF and its function to block the combination with VEGFR.