王志雄等

[摘要] 目的 研究桑叶对肿瘤血管生成中血管内皮细胞增殖、迁移和管腔形成的影响。 方法 用CCK-8试剂盒测定HUVEC-2C细胞数量的变化；用Transwell转移小室研究HUVEC-2C细胞迁移情况；用包被Matrigel的24孔板来培养HUVEC-2C细胞观察其管腔形成情况。实验分为实验组和对照组，每组分别设置3个复孔，在实验组中加入桑叶浸出液。观察桑叶对肿瘤血管生成的影响。 结果 桑叶液可抑制HUVEC-2C细胞的增殖能力，抑制作用随着浓度的增长而增强。在浓度为100、50、25 mg/mL时，与对照组比较，差异有统计学意义（P < 0.05）。当浓度为100 mg/mL的桑叶液时细胞增殖抑制率可达到37.6%。通过观察Transwell小室，与对照组比较，添加了桑叶液的HUVEC-2C细胞，其迁移能力受到一定的制约作用。Matrigel实验中，与未加入桑叶液的HUVEC-2C细胞所形成的较完整的环状结构比较，加入桑叶液的HUVEC-2C细胞在管腔形成方面受到明显的抑制作用。 结论 桑叶液通过抑制血管内皮细胞增殖、迁移和管腔形成，对肿瘤血管的生成具有抑制作用。

[关键词] 桑叶；肿瘤血管；内皮细胞；增殖；迁移；管腔形成

[中图分类号] R282.71 [文献标识码] A [文章编号] 1673-7210（2014）09（a）-0022-04

[Abstract] Objective To study the influence of mulberry on vascular endothelial cell proliferation and migration as well as lumen forming in tumor angiogenesis. Methods The change of HUVEC-2C cell quantity was measured with CCK-8 kit； HUVEC-2C cell migration was studied in Transwell chamber； HUVEC-2C cells were cultured in a 24-pore plate coated with Matrigel to observe its lumen forming. The experiment consisted of experiment group and control group， with three complex holes set for each group. Mulberry leachate was added to the experiment group. The effect of mulberry leaf to tumor angiogenesis was observed. Results Mulberry leachate could inhibit the proliferation ability of HUVEC-2C cell， with the inhibiting effect strengthened as the concentration increases. When the concentration was 100， 50， 25 mg/mL， the difference between the experiment group and the control group had statistical significance （P < 0.05）. When the concentration was 100 mg/mL， the cell proliferation inhibiting ratio of mulberry leachate could reach 37.6%. Observation on Transwell chamber showed the follows： added with mulberry leachate， the HUVEC-2C cells′migration ability was inhibited to some extent compared with the control group. In Matrigel experiment， compared with the comparatively complete cyclic structure formed by HUVEC-2C cells without mulberry leachate， the HUVEC-2C cells added with mulberry leachate were obviously inhibited in term of lumen forming. Conclusion Mulberry leachate has inhibiting effect on tumor angiogenesis by inhibiting vascular endothelial cell proliferation and migration as well as lumen forming.

[Key words] Mulberry leaf； Tumor angiogenesis； Endothelial cells； Proliferation； Migration； Lumen formation

1971年，Folkman[1]首次提出血管生成与肿瘤生长之间的关系问题。随后许多研究陆续证明，肿瘤形成初期，肿瘤细胞能够通过弥散方式获取营养物质。但长到一定大小后，必须从最近的血管处诱导形成自身血管。如果没有血管生成这一过程，肿瘤就会因得不到足够的营养和氧气而进入休眠阶段，甚至坏死[1]。血管为肿瘤生长提供营养和氧气，也是其排泄的通道，更是转移的必要条件[2]。这就为治疗肿瘤提供了一个新思路——通过阻断肿瘤血管的生长，切断肿瘤营养与氧气的供应，从而抑制肿瘤的发生和发展。肿瘤血管与正常血管存在很大差异，但无论是肿瘤血管还是正常的血管，血管内皮细胞（endothelial cell）都是血管的主要组成部分，衬贴于血管内壁。

目前，市场上销售的血管生成抑制药物大都成本高昂，作用途径比较单一，治疗不够彻底，副作用仍然较大，在实际应用中存在一些问题。……