香菇多糖体外抗血管生成作用实验研究

Study on the Anti-Angiogenic Effects of Lentinan in Vitro Experiment

目的:研究香菇多糖对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)增殖、迁移、黏附以及体外血管生成作用的影响。方法:香菇多糖设31.25 mg·L-1、62.5 mg·L-1、125 mg·L-1、250 mg·L-1、500 mg·L-1等5个质量浓度,以沙利度胺(200 mg·L-1)为阳性对照。采用MTT法检测香菇多糖对HUVEC细胞增殖的影响,Transwell小室迁移实验检测其对细胞迁移的影响,纤维连接蛋白(Fn)黏附实验检测其对细胞黏附的影响,Tuber formation assay法检测其对体外血管生成的影响。结果:经香菇多糖干预后,HUVEC细胞增殖、迁移、黏附及血管生成均受到明显抑制。当质量浓度大于62.5 mg·L-1时与对照组比较,差异有统计学意义。随着浓度的增加,抑制作用逐渐增强,质量浓度为250 mg·L-1时对细胞增殖的抑制率最高,为43.8%。香菇多糖抑制细胞迁移、黏附和血管生成的作用呈剂量依赖性,质量浓度为500mg·L-1时抑制作用最显著。结论:香菇多糖可显著抑制HUVEC的增殖、迁移、黏附以及体外血管生成,提示香菇多糖抗肿瘤作用可能与抑制血管生成有关。

Objective： To study the effects of lentinan on the cell proliferation,migration,adhesion and angiogenesis in virto of human umbilical vein endothelial cell（ HUVEC）. Methods： Lentinan were prepared 5 concentrations as 31. 25,62. 5,125,250 and 500 mg·L-1,and thalidomide（ 200 mg·L-1） was used as positive control. MTT method was used to detect the effects of lentinan on the proliferation of HUVEC,Transwell Chambers migration experiment was applied to test its influence on cell migration,fibronectin（ Fn） adhesion experiment was applied to test its influence on cell adhesion,Tuber formation assay method was used to detect their effects on angiogenesis in vitro. Results： After lentinan intervention,cell proliferation,migration,adhesion and angiogenesis of HUVEC had been significantly inhibited. Compared with the control group,the difference was statistically significant when the concentration of lentinan is greater than 62. 5 mg·L-1. And with the increase of concentration,its inhibition on cell proliferation enhanced gradually,and reached the highest inhibition ratio of 43. 8% when the concentration was 250 mg·L-1. Inhibiting effects of lentinan on cell migration,adhesion,and angiogenesis presented dose-dependent,had the most significantly inhibitory effect at the concentration of 500 mg·L-1. Conclusion： Lentinan could significantly inhibit cell proliferation,migration,adhesion,and angiogenesis in vitro of HUVEC,which suggested the anti-tumor effects of letinan may be related with inhibiting angiogenesis.