黄芪甲苷改善人内皮祖细胞生物学功能的实验研究(1)

http://www.100md.com 2020年1月1日 《湖南中医药大学学报》 20201

     〔摘要〕 目的 探讨黄芪甲苷(Astragaloside IV，AS-IV)对人内皮祖细胞(endothelial progenitor cells，EPCs)生物学功能的影响，为深入研究AS-IV调节EPCs介导的血管新生的作用机制奠定基础。方法 取足月健康新生儿脐带血，采用密度梯度离心法分离得到单个核细胞，经传代培养，采用CD31抗体联合DAPI核染以及FITC-UEA-I和 Dil-ac-LDL双荧光染色法鉴定EPCs 。将鉴定成功的EPCs随机分为实验组和对照组，实验组采用100 mg/L的AS-IV干预，对照组用等容量的PBS液处理，两组细胞培养24 h后，通过CCK-8细胞增殖检测试剂盒、黏附能力测定试验、细胞划痕试验及Matrigel体外成血管试验观察AS-IV对EPCs增殖、黏附、迁移和成管功能的影响。结果 与对照组相比，实验组细胞增殖的OD值增大，细胞黏附数增多，细胞迁移宽度增加(即细胞迁移率增大)，体外成管数增多，差异均有统计学意义(P<0.05)。结论 黄芪甲苷能改善体外人EPCs的生物学功能，具有调节EPCs介导血管新生的潜能。

    〔关键词〕 黄芪甲苷;内皮祖细胞;生物学功能;血管新生

    〔中图分类号〕R285.5 〔文献标志码〕A 〔文章编号〕doi：10.3969/j.issn.1674-070X.2020.01.008

    〔Abstract〕 Objective To explore the effects of Astragaloside IV (AS-IV) on the biological function of human endothelial progenitor cells (EPCs)， and to lay a foundation for further study of the mechanism of AS-IV regulating EPCs-mediated angiogenesis. Methods Umbilical cord blood was collected for full-term healthy newborns， and the mononuclear cells were separated by density gradient centrifugation method. After subculture， CD31 antibody combined with DAPI staining identification， and double staining identification of FITC-UEA-I and Dil-ac-LDL were adopted to identify EPCs. The successfully identified EPCs were randomly divided into an experimental group and a control group. The experimental group was treated with 100 mg/L Astragaloside IV and the control group was treated with the same amount of PBS solution. After 24 hours of cell culture， the effects of AS IV on EPCs proliferation， adhesion， migration and tube formation were observed by CCK-8 cell proliferation kit， adhesion ability assay test， cell scratch test and Matrigel angiogenesis test in vitro. Results Compared with the control group， the OD value of cell proliferation， the number of cell adhesion， the width of cell migration (namely cell migration rate) and the number of tubes formed in vitro increased in the experimental group， with significant difference (P<0.05). Conclusion Astragaloside IV can improve the biological function of human EPCs in vitro and has the potential to regulate the angiogenesis mediated by EPCs.

    〔Keywords〕 Astragaloside IV; endothelial progenitor cells; biological function; angiogenesis

    內皮祖细胞(endothelial progenitor cells，EPCs)是一类具有游走特性的能自我增殖更新和分化为血管内皮细胞的定向干细胞。1997年Asahara等[1]利用免疫磁珠细胞分选法从人外周血中成功分离得到EPCs。随着对EPCs分离、培养、鉴定、功能和应用方面的研究的广泛开展，为难治性缺血性疾病的治疗提供了新思路。而这与EPCs增殖、黏附、迁移和成管等生物学功能密不可分，如何通过药物或物理疗法诱导EPCs以提高其生物学功能将有助于EPCs发挥血管新生作用。黄芪甲苷(Astragaloside IV，AS-IV)是中药黄芪中最主要的活性成分，具有血管药理活性，能诱导血管新生[2]。本团队前期研究表明AS-IV能提升EPCs的细胞活性，且发现AS-IV促人EPCs增殖的最适浓度为100 mg/L[3]。本研究是在前期研究基础上进一步探讨AS-IV对EPCs增殖、黏附、迁移和成管等生物学功能的影响，将为深入研究AS-IV作用于EPCs介导的血管新生奠定基础。(蔡昫 邹晓玲 王禹萌 余亦程 兰宏伟 郭娟 王婷婷 熊武)

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