巴戟天糖链对血管内皮祖细胞增殖分化和旁分泌的作用

Effects of morinda officinalis oligosaccharide on the proliferation,differentiation and paracrine of vascular endothelial progenitor cells

背景:巴戟天糖链是巴戟天醇提物的主要有效成分,其促进缺血区组织的血管生成作用十分明确,而机制不明。目前认为血管内皮的修复主有两种方式:血管内皮细胞分裂而来或者是由末梢血液中具有分化功能的血管内皮祖细胞分化而来。实验通过研究巴戟天糖链与血管内皮祖细胞生物学功能是否存在联系,解释巴戟天糖链促进血管生成机制,为研发药物提供实验证据。目的:探讨巴戟天糖链对血管内皮祖细胞增殖分化和旁分泌的影响。方法:从健康人的外周血中分离培养血管内皮祖细胞,将血管内皮祖细胞分为对照组(不含巴戟天糖链)和实验组(含质量浓度为0.15 g/L巴戟天糖链),在体外培养48 h,通过荧光染料染色检测血管内皮祖细胞的增殖情况,流式细胞仪检测血管内皮祖细胞中CD31阳性表达比例,酶联免疫吸附法检测培养基中血管内皮生长因子、基质细胞衍生因子1和白细胞介素8水平。结果与结论:①血管内皮祖细胞表面标志物CD34、CD133和VEGFR-2均为阳性的比例为(84.72±4.34)%;②经过质量浓度为0.15 g/L巴戟天糖链培养48 h后,实验组中血管内皮组细胞增殖率和CD31阳性表达比例明显高于对照组(P<0.05);③实验组中血管内皮生长因子、基质细胞衍生因子1和白细胞介素8水平明显高于对照组(P<0.05);④结果表明,巴戟天糖链可以促进血管内皮祖细胞的增殖和分化,同时还可以诱导血管内皮祖细胞通过旁分泌途径释放血管内皮生长因子、基质细胞衍生因子1和白细胞介素8等细胞活性因子,对于心肌缺血性疾病具有潜在的治疗价值。

BACKGROUND:Morinda officinalis oligosaccharide is the main active ingredient of morinda officinalis extract which can promote the angiogenesis of ischemic tissue,but the mechanism is unknown.At present,there are two ways for endothelial repair:vascular endothelial cell division or differentiation from vascular endothelial progenitor cells in the peripheral blood.Here,we attempted to explain the pro-angiogenesis mechanism of morinda officinalis oligosaccharide by exploring whether there is a correlation between morinda officinalis oligosaccharide and the biological function of vascular endothelial progenitor cells,thereby providing experimental reference for new drug development.OBJECTIVE:To observe the effect of morindae officinalis oligosaccharide on the proliferation,differentiation and paracrine of vascular endothelial progenitor cells.METHODS:Vascular endothelial progenitor cells were isolated from healthy human peripheral blood,and divided into two groups:control group(without morindae officinalis oligosaccharide)and experimental group(with 0.15 g/L morindae officinalis oligosaccharide),followed by 48 hours of in vitro culture.The proliferation of vascular endothelial progenitor cells was tested by fluorescent staining;the ratio of vascular endothelial progenitor cells expressing CD31 was detected by flow cytometry;and the levels of vascular endothelial growth factor,stromal cell-derived factor 1 and interleukin 8 were analyzed by enzyme-linked immunosorbent method.RESULTS AND CONCLUSION:The percentage of vascular endothelial cells expressing CD34,CD133 or VEGFR-2 was(84.72±4.34)%.After 48 hours of culture by 0.15 g/L morindae officinalis oligosaccharide,the proliferation rate and the positive expression of CD31 in the experimental group were significantly higher than those in the control group(P<0.05).The levels of vascular endothelial growth factor,stromal cell-derived factor 1 and interleukin 8 in the experimental group were also higher than those in the control group(P<0.05).To conclude,morindae officinalis oligosaccharide can promote the proliferation and differentiation of vascular endothelial progenitor cells,and meanwhile,it can stimulate the release of vascular endothelial growth factor,stromal cell-derived factor 1 and interleukin 8 from vascular endothelial progenitor cells through the paracrine pathway.Consequently,it is a potential drug for myocardial ischemic diseases.