【目的】研究柔毛水杨梅对人微血管内皮细胞增殖、迁移、成管能力的影响。【方法】将不同浓度(0~200 ng/m L)柔毛水杨梅作用于人微血管内皮细胞(HMVEC-Ls),分别采用MTS法检测细胞的增殖活性,划痕法检测细胞的迁移能力,Matrigel法检测细胞的成管能力以及Western blot法验证低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、端粒酶逆转录酶(TERT)的表达水平。【结果】MTS结果显示:柔毛水杨梅与HMVEC-Ls共同培养7 d,25~200 ng/m L不同浓度组均可促进HMVEC-Ls增殖,与对照组(0 ng/m L)比较,差异均有统计学意义(P〈0.05),以100 ng/m L组促进HMVEC-Ls增殖能力最显著(P〈0.05)。划痕实验结果显示:与对照组比较,50~150 ng/m L组HMVEC-Ls迁移率显著增大,差异均有统计学意义(P〈0.05),以150 ng/m L组HMVEC-Ls迁移率最高。成管实验结果:与对照组比较,50~150 ng/m L组HMVECLs成管长度显著增加,差异均有统计学意义(P〈0.05),亦以150 ng/m L组HMVEC-Ls成管长度最大。Western blot检测结果:与对照组比较,150 ng/m L柔毛水杨梅浓度组HIF-1α、TERT和VEGF表达均显著升高(P〈0.05)。【结论】柔毛水杨梅可通过促进人微血管内皮细胞增殖、迁移、管样形成而发挥促血管新生效应,其机制可能与HIF-1α-TERT-VEGF途径介导的血管新生有关。
Objective To study the effects of Geum japonicum ThuMb. var. chinense F. Bolle (GJ) on the proliferation, migration and tube formation of human microvascular endothelial cells (HMVEC-Ls) in vitro. Methods The HMVEC-Ls were treated with different concentrations of GJ methanol extract. HMVEC-Ls proliferation, migration and tube formation were measured by MTS assay, wound-healing cell migration assay and Matrigel assay, respeetively. The effect of GJ methanol extract on the expression levels of hypoxia inducible factor 1 α( HIF- 1α), telomerase reverse transcriptasc (TERT) and vascular endothelial growth factor( VEGF )were detected by Western blotting method. Results MTS results showed that HMVEC-Ls proliferation index in 25- 200 ng/mL of GJ groups was increased(P〈0.05 as compared with that of the control group) , the effect of 100 ng/mL of GJ group being the best. HMVEC-Ls migration ratio in 50-150 ng/mL of GJ groups was enhanced(P〈 0.05 as compared with that of the control group), the effect of 150 ng/mL of GJ group being the best. The tube length of HMVEC-Ls in 50-150 ng/mL of GJ groups was enhanced (P〈0.05 as compared with that of the control group), the effect of 150 ng/mL of GJ group being the best. In addition, HIF-1α, TERT and VEGF protein expression levels were also significantly up-regulated by GJ (P〈0.05). Conclusion GJ can promote HMVEC-Ls proliferation, migration and tube formation, and the mechanism is related with angiogenesis mediated by HIF-1α-TERT-VEGF pathway.