补阳还五汤对外周血内皮祖细胞数量和功能的影响

背景:现代医学研究表明,内皮祖细胞和补阳还五汤在治疗缺血性疾病上具有相似的功能,两者之间是否存在着一定的联系?目的:观察补阳还五汤对外周血内皮祖细胞数量和功能的影响。方法:采用密度梯度离心法分离培养兔外周血内皮祖细胞,经Dil-ac-LDL和FITC-UEA-I双染色和表面抗原的免疫组织化学检测鉴定后,将贴壁细胞随机分为4组,分别用基础培养基以及含有20,50,100g/L补阳还五汤的EBM-2培养基进行细胞培养72h。检测细胞形态及计数,再采用四甲基偶氮唑盐微量酶反应比色法、Transwell小室、黏附功能检测、一氧化氮检测及血管内皮生长因子免疫细胞化学分析来评定其增殖、迁移、黏附、分泌一氧化氮和表达血管内皮生长因子的能力。结果与结论:不同质量浓度补阳还五汤均能增加内皮祖细胞的数量,提高内皮祖细胞增殖、黏附、迁移和分泌一氧化氮的能力(P<0.05),药物质量浓度在50g/L时影响最为显著(P<0.01),而对血管内皮生长因子表达的影响较微弱。提示补阳还五汤能增加内皮祖细胞的数量,提高内皮祖细胞的增殖、迁移和黏附能力,并可能诱导晚期内皮祖细胞的分化。

Effect of Buyang Huanwu decoction on number and function of endothelial progenitor cells in the peripheral blood

BACKGROUND:Modern medicine studies have confirmed that endothelial progenitor cells(EPCs) and Buyang Huanwu Decoction(BHD) have similar functions for treatment of ischemia disease.Is there a correlation between them? OBJECTIVE:To explore the effects of BHD on the number and function of EPCs in peripheral blood.METHODS:EPCs were isolated from rabbit peripheral blood by density gradient centrifugation.Cultured EPCs were identified as Dil-LDL uptaking and FITC-lectin binding and further demonstrated using surface antigen immunohistochemistry.The attached cells were randomized into four groups.After being cultured in basal medium and EBM-2 medium containing different concentrations of BHD extract(20,50,100 g/L) for 72 hours,the cell morphology was observed.Meanwhile,the proliferation,migration,and adhesion of EPCs were assayed with MTT,Transwell chamber and adhesion function test,respectively.In addition,nitric oxide content and vascular endothelial growth factor(VEGF) expression were assessed.RESULTS AND CONCLUSION:Various mass concentrations of BHD could increase the number of EPCs,elevated proliferation,adherence,migration and nitric oxide secretion capacity of EPCs(P < 0.05).The effect was most significant at 50 g/L(P < 0.01).However,the effect was weak on VEGF expression.Results indicated that BHD could significantly increase the number of EPCs,and enhance the proliferation,migration,and adhesive capacity of EPCs.Meanwhile,BHD can induce late EPCs differentiation.