丹酚酸B对EPCs释放VEGF、SDF-1、IL-8和MMP9细胞因子及EPCs黏附能力的影响

Effect of salveanolic acid B on the release of VEGF,SDF-1,IL-8 and MMP9 cytokines and adhesion ability of EPCs

目的研究丹酚酸B对血管内皮祖细胞黏附和旁分泌功能的影响。方法从健康人的外周血中分离培养血管内皮祖细胞,鉴定后将其分为对照组(不含丹酚酸B体),20、40、60μg/mL丹酚酸B组,分组后细胞培养48 h,CCK8试剂盒检测血管内皮祖细胞的增殖情况;流动腔实验检测血管内皮祖细胞黏附数量;酶联免疫吸附法检测培养基中细胞因子VEGF、SDF-1、IL-8和MMP9的浓度。结果体外培养7 d后检测血管内皮祖细胞表面标志物CD34、CD45和VEGRF-2,其中内皮祖细胞比例为89.11%±6.09%;不同浓度丹酚酸B组血管内皮祖细胞培养2 d后,细胞增殖比例与对照组比较差异有统计学意义(P<0.05);流动腔实验结果显示,20、40、60μg/mL丹酚酸B组每个低倍镜视野的细胞计数分别为235±24、393±28和551±35,与对照组(98±13)比较差异有统计学意义(P<0.05);不同浓度丹酚酸B培养4 d后,不同浓度丹酚酸B组培养基中VEGF、SDF-1、IL-8和MMP9浓度与对照组比较差异有统计学意义(P<0.05)。结论丹酚酸B可以促进血管内皮祖细胞增殖,增强内皮祖细胞的黏附功能,还可以通过旁分泌途径释放VEGF、SDF-1、IL-8和MMP9。

Objective To observe the effect of salveanolic acid B on function of the adhesion and paracrine of endothelial progenitor cells. Methods Endothelial progenitor cells were collected from healthy human peripheral blood. After identification, the endothelial progenitor cells were divided into control group （ without salveanolic acid B ） ,20,40 and 60 μg/mL salveanolic acid B groups. At 48 h, the proliferation of endothelial progenitor cells was tested by CCK8 kit;endothelial cell number was counted by flow cavity tests ; VEGF, SDF-1, IL-8 and MMP9 were analyzed by ELISA method. Results The ratio of the vascular endothelial cells expressing CD34, CD45 and VEGRF-2 together was 89. 11% ± 6. 09 % after 7 d cultured in vitro ; after being cultured under different concentration of salveanolic acid B for 2 d, compared with control group, the proliferation rates in salveanolic acid B groups were significantly different （ P 〈 0. 05 ） ;flow chamber test showed that cell counts in different salveanolic acid B groups were 235 ± 24,393 ± 28 and 551 ± 35, and compared with control group （ 98 ± 13 ） , the difference was statistically significant （ P 〈 0. 05 ） ; after be- ing cultured under different concentrations of salveanolic acid B for 4 d, compared with control group, the concentra- tions of VEGF, SDF-1,IL-8 and MMP9 in different salveanolic acid B groups were significantly different （P 〈 0. 05 ）. Conclusion The salveanolic acid B can promote the proliferation and adhesion of endothelial progenitor cells and stimulate endothelial progenitor cells to release VEGF, SDF-1 ,IL-8 and MMP9 through paracrine.