摘要
目的:观察丹参对高胆固醇血症患者内皮祖细胞(endothelial progenitor cells,EPCs)的保护作用。方法:年龄性别匹配的健康志愿者及高胆固醇血症患者各6例入选研究,各取空腹外周血20 mL,采用密度梯度离心法分离单个核细胞,贴壁选择法行EPCs培养,以健康志愿者为对照组,各高胆固醇血症患者血样一分为二,分别为高胆固醇组及丹参组,丹参组培养时另添加复方丹参注射液10μg/mL,采用流式细胞术、倒置显微镜、MTT法、Boyden小室等分别观察各组EPCs数量、集落形成能力、增殖能力、黏附能力及迁移能力。结果:高胆固醇血症患者外周血EPCs数量较对照组显著下降〔(6.13±0.84)vs(24.53±3.67),P<0.01〕,且其克隆形成能力、增殖能力、黏附能力及迁移能力也显著降低,培养时添加复方丹参注射液使丹参组克隆形成能力、增殖能力、黏附能力及迁移能力较高胆固醇组显著提高,但低于对照组,其中黏附能力及迁移能力虽仍较对照组低,但无统计学差异(P>0.05)。结论:高胆固醇血症患者外周血内皮祖细胞数量下降,功能受损,复方丹参注射液对高胆固醇血症患者外周血培养的内皮祖细胞功能有保护作用。
Objective ： To investigate the protective effects of Danshen on number and activity of endothelial progenitor cells （EPCs） of patients with Hypercholesterolemia. Method： 6 healthy volunteer and 6 patients with Hypercholesterolemia were enrolled in the study. 20 mL peripheral blood was obtained from each object ,The blood sample of patients with Hypercholesterolemia were divided into two parts. Mononuclear fraction of human peripheral blood was collected by density gradient centrifugation, plated on fibronectin coated culture dishes. The dishes were divided into three groups： control group, and Hypercholesterolemia group and Danshen group（ receiving Denshen 10 μg/mL in culture media） . Cell clusters were viewed with an inverted microscope, fluorescence -activated cell sorting （FACS） analysis of PE - CD34 and FITC - AC133 was performed to detect number of EPCs, EPC proliferation and migration were assayed with MTT assay, modified Boyden chamber assay. EPCs adhesion ability assay was performed by replafing cells on fibronectin - coated dishes, and then counting adherent cells. Results： Numbers of EPCs （10^3cells per 1 mL peripheral blood）of Hypereholesterolemia group was lower than control group[ （6.13±0.84） vs （24.53±3. 67）, P 〈0.01 ]. Compared with group control, numbers of clusters （ per 40 power microscopic field ） , OD value, adhesive EPCs （ per 400 power microscopic field） and migratory EPCs（ per 200 power microscopic field） of Hypercholesterolemia group were. significantly decreased （4.56 1.11 vs 15.57 2.59, P 〈0.01,0.19 0.02 vs 0.29 0.04, P 〈0.05,8.20 0.75 vs 20.61 8.41, P 〈0.01 and 13.18 1.17 vs 22.277.98; P 〈0.01 ,respectively）. After treated with Denshen 10 ug/mL in culture media, numbers of clusters, OD value, adhesive EPCs and migratory EPCs of Danshen group were significantly decreased （compared with Hypercholesterolemia group ： 11.42 2.03 vs 4.56 1.11, P 〈 0.01,0.23 0.02 vs 0.19 0.02, P 〈 0.05, 14.79 1.93 vs 8.20 0.75, P 〈0.01 andl
出处
《中华中医药学刊》
CAS
2007年第6期1173-1175,共3页
Chinese Archives of Traditional Chinese Medicine
基金
温州市科技局基金项目(Y2005B031)
浙江省自然科学基金项目(M303648)
关键词
丹参
内皮祖细胞
细胞培养
高胆固醇血症
Danshen
endothelial progenitor cells
cell culture
Hypereholesterolemia
