骨化三醇对体外培养内皮祖细胞的药理作用

Pharmacological effects of calcitriol on endothelial progenitor cells in vitro

目的观察骨化三醇[1,25-(OH)2D3]干预培养对人外周血来源的内皮祖细胞(Endothelial progenitor cells,EPCs)数量及功能的影响。方法采用密度梯度离心法和差速贴壁法,获得EPCs细胞,用流式细胞仪和激光共聚焦显微镜进行鉴定;然后分别加入不同浓度的骨化三醇10、50、100μmol/L干预培养72 h,观察EPCs的数量变化、黏附、增殖和产生一氧化氮(NO)的能力。结果与对照组相比,10、50、100μmol/L组的贴壁细胞个数分别为13.91±1.58、14.28±1.66、16.91±1.85,差异有统计学意义(P<0.01);重贴壁细胞个数分别为9.55±1.11、10.25±1.15、10.88±1.02,差异有统计学意义(P<0.01);增殖能力(OD值)分别为0.285±0.005、0.301±0.009、0.321±0.006,差异无统计学意义(P>0.05);培养液中NO含量分别为5.687 9±0.392 5、5.954 7±0.243 2、7.682 2±0.353 4μmol/L,差异无统计学意义(P>0.05)。结论骨化三醇干预培养能提高人外周血来源EPCs的黏附能力,但对其增殖能力和产NO能力无明显影响。

Objective To observe the effect of calcitriol [ 1,25- （OH） 2 D3 ] on the number and function of en- dothelial progenitor cells （EPCs） in human peripheral blood. Methods EPCs were obtained by Ficoll density gradient centrifugation and differential adherence method, and identified by flow cytornetry and confocal laser scanning micros- copy. Then, the attached cells were stimulated with calciriol （ 10,50 and 100 p, mol/L） for 72 h. The number of EPCs, adhesion,proliferation, and production of nitric oxide （NO） were observed. Results Compared with control group, the number of attached cells in 10,50 and 100 p, mol/L groups were 13.91 ± 1.58,14. 28± 1.66, and 16. 91 ± 1.85 ,respec- tively,the difference being statistically significant （P 〈 0. 01 ） ; the number of reattached cells were 9. 55 ± 1. 11, 10. 25 ± 1. 15 and 10. 88 ± 1.02, respectively, the difference being statistically significant （ P 〈 0. 01 ） ; proliferation （OD value） were 0. 285 ± 0. 005,0. 301 ± 0. 009 and 0. 321 ± 0. 006, respectively, and there was no statistically signifi- cant difference （P 〉0.05）;the NO content in culture medium was 5. 687 9 ±0.392 5,5. 954 7 ±0.243 2 and 7. 682 2 ±7. 353 4 μmol/L, respectively, there being no statistically significant difference （ P 〉 0. 05 ）. Conclusion Calcitriol can improve adhesive ability of EPCs,but has no effect on either proliferation or secretion of NO in EPCs in vitro.