高良姜素的体内外肿瘤血管生成抑制作用

The in vitro and in vivo antiangiogenenic effect of galangin

目的探讨高良姜素体内、体外对肿瘤血管生成的影响。方法采用磺酰罗丹明B(SRB)法测定高良姜素对人微血管内皮细胞(EaHy926)增殖的影响;细胞划痕实验检测其对EaHy926迁移能力的影响;观察其对鸡胚绒毛尿囊膜(CAM)模型体内血管生成的影响。实验分为对照组(高良姜素0μg)、高良姜素药物组(1、5和10μg);采用H22细胞株建立裸鼠肝癌种植瘤模型,模型小鼠分成5组,阴性对照组,高良姜素低、中、高剂量组小鼠按照10、20和40 mg/(kg·d)分别灌胃给药,阳性对照组环磷酰胺按20 mg/(kg·d)腹腔注射,连续15 d;免疫组化法分析肿瘤组织中微血管密度(MVD),观察高良姜素裸鼠对肿瘤生长及肿瘤血管形成的影响。结果不同浓度的高良姜素对EaHy926细胞生长、迁移能力及体外Matrigel小管的形成均有抑制作用,且抑制作用随药物浓度递增而增强;高良姜素5和10μg组对CAM血管形成均呈现较强的抑制作用(P<0.05,P<0.01);与模型组相比,高良姜素20和40 mg/(kg·d)剂量组和环磷酰胺组能显著降低肿瘤瘤重(P<0.05),其抑瘤率分别为34.17%,79.73%,55.71%;同时,高良姜素20和40 mg/(kg·d)剂量组能显著降低肿瘤组织中MVD(P<0.05)。结论高良姜素体内体外都具有抑制肿瘤血管生成的作用。

Objective To investigate the antiangiogenic effect of galangin in vitro and in vivo. Methods The inhibitory effect of galangin on human umbilical vein endothelial cells（EaHy926）was tested by sulphorhodamine（SRB）method,the EaHy926 endothelial cell migration was assessed using in vitro model system,and the in vivo antiangiogenic effect of galangin at 1,5 and 10 μg was evaluated using the chorioallantoic membrane（CAM）model. The H22 tumor-bearing model was established using BALB/c nude mice,which were divided into five groups：The model group,positive control group[ip cyclophosphamide 20 mg/（kg·d）]and the galangin 10,20 and 40 mg/（kg·d）groups,respectively. After the daily administration for 15 consecutive days,the tumors grown in the nude mice were examined and the microvessel density（MVD）was tested via examining the expression of CD31 in the tumor tissues by immunohistochemistry in order to evaluate the effect of galangin to the tumor growth and the angiogenesis of the tumors. Result Galangin inhibited both the growth,migration and Matrigal tubule of EaHy926 cells in a dose-dependent manner in the in vitro test.Further in the in vivo test,galangin could also obviously inhibit the angiogenesis of CAM at the 5 and 10 μg concentration. The tumor mass and the relative tumor volume in the 20 and 40 mg/（kg·d）galangin groups and the positive control cyclophosphamide group were significantly lower（P〈0.05）than those in the model group in the in vivo nude mice test. The tumor inhibitory rates of those three groups were 34.17%,79.73% and 55.75%,respectively. The MVD was also significantly lower in the high dosage galangin groups than that in the model group. Conclusion Galangin showed the obvious anti-angiogenenic effect both in vitro and in vivo in the present study.