疏肝活血方含药血清对内皮祖细胞数量及功能的影响  

Effect of Shugan Huoxue(疏肝活血) Recipe on Number and Activity of Endothelial Progenitor Cells

作　　者：李大勇[1] 陈文娜[2] 谷峰[2] 郑巧楠[1] 袁明殿[1] 吕延伟 

机构地区：[1]辽宁中医药大学附属医院血管外科,沈阳110032 [2]辽宁中医药大学

出　　处：《中国中西医结合外科杂志》2009年第5期535-539,共5页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助(30600843);辽宁省科技厅博士启动基金资助(20061030)

摘　　要：目的:观察疏肝活血方含药血清对大鼠骨髓来源的血管内皮祖细胞(EPCs)数量及功能的影响。方法:从大鼠骨髓中获取单个核细胞,体外培养9d后,收集贴壁细胞,加入低、中、高浓度的疏肝活血方及其拆方的含药血清,培养12、24、48h。激光共聚焦显微镜鉴定,FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,并在倒置荧光显微镜下计数,分别采用黏附能力测定、MTT比色法及改良的Boyden小室实验来观察疏肝活血中药对EPCs黏附能力、增殖能力和迁移能力的影响,用流式细胞仪检测PI-/AnnexinⅤ+的凋亡细胞。结果:疏肝活血方能明显促进EPCs的分化,提高EPCs的数量,明显改善EPCs的黏附、增殖及迁移能力,减少EPCs的凋亡率,中浓度组培养24h达到作用的高峰(P<0.01)。结论:疏肝活血方能促进EPCs数量的增加、功能的改善,减少EPCs的凋亡。Objective To investigate the effects of Shugan Huoxue（疏肝活血） recipe on the number and activity of endothelial progenitor cells （EPCs） derived from rat bone marrow. Methods Total mononuclear cells were isolated from rat bone marrow.After 9-day culture,the attached cells were stimulated with the drug serum of Shugan Huoxue recipe in series of final concentrations,and its separated recipe for 12,24,and 48 h.EPCs were assessed by laser scanning confocal microscopy,FITC-UEA-I and Dil-acLDL were both positive,were the differentiation of EPCs,and calculated by inverted fluorescent microscopy.EPCs adhesion assay was performed by replacing those on fibronectin-coated dishes,then adherent cells were counted.EPCs proliferation and migration were assayed with MTT assay and modified Boyden chamber assay respectively.The apoptosis of EPCs were identified by flow cytometry,and EPCs that are in early apoptosis are Annexin-V positive and PI negative. Results Shugan Huoxue recipe promoted the differentiation of EPCs increased the number of EPCs,promoted adhesive proliferative,and migratory capacity of EPCs,and decreased the apoptosis of EPCs,maximum at 24 hours in the middle concentration （P〈0.01）. Conclusion Shugan Huoxue recipe can promote EPCs augmentation,enhance its functional activity,and decrease the apoptosis of EPCs.

关 键 词：疏肝活血方 内皮祖细胞 治疗性血管生成 血清药理学 

分 类 号：R285.5[医药卫生—中药学]