双氢青蒿素对小鼠淋巴管内皮细胞增殖与迁移及管样结构形成的影响

Effect of Dihydroartemisinin on the Proliferation,Migration and Tube-like Structure Formation of Mouse Lymphatic Endothelial Cells

目的探讨双氢青蒿素(DHA)对小鼠淋巴管内皮细胞(LEC)增殖、迁移和管样结构形成的影响。方法腹腔注射不完全弗氏佐剂诱导小鼠淋巴管瘤形成,分离并培养小鼠LEC。采用MTT法检测DHA对LEC增殖的抑制作用,流式细胞仪检测LEC的凋亡率,Transwell迁移实验观察DHA对LEC迁移的影响;基质胶实验检测DHA对LEC管样结构形成的影响,实时定量RT-PCR和Western-blot检测DHA处理后LEC表达VEGFR-3 mRNA和蛋白的情况。结果5~40μg.mL-1DHA能显著抑制LEC增殖,并诱导不同程度的凋亡,且具有剂量效应关系。Transwell迁移实验和基质胶实验分别显示DHA能抑制LEC迁移和体外自发性管样结构形成。DHA处理之后,LEC表达VEGFR-3 mRNA和蛋白均下调。结论DHA能抑制LEC增殖、迁移和管样结构形成,诱导LEC凋亡和下调VEGFR-3表达,有望成为拮抗淋巴管生成的药物。

Objective To explore the effect of antimalarial dihydroartemisinin （DHA） on the proliferation, migration and tube-llke structure formation of mouse lymphatic endothelial cells （LEC）. Methods Benign lymphangiomas were induced by intraperitoneally injected emulsified incomplete Freund^s adjuvant and mechanically disrupted to obtain LEC. MTT assay was used to observe the inhibitory effect of DHA on the growth of LEC. Apoptosis rate of LEC was evaluated by flow eytometry. Migration and tube-like structure formation of LEC were detected by transwell migration assay and matrigel assay, respectively. The expressions of VEGFR-3 mRNA and protein in LEC by DHA were detected by real time quantitative RT-PCR and Western-blot, respectively. Results DHA at 5 - 40 μg · mL^-1 significantly inhibited LEC proliferation and induced apoptosis in a dose-dependent manner. Cell migration and tube-like structure formation of LEC were suppressed by DHA. Treated with DHA, the expression of VEGFR-3 mRNA and protein of LEC were down-regulated. Conclusion DHA can inhibit the proliferation, migration and tube-like structure formation of mouse LEC, induce the apoptosis of LEC, and down-regulate the expression of VEGFR-3. Our results suggest that DHA might be a candidate as an anti-lymphangiogenesis inhibitor.