维生素D对活动期系统性红斑狼疮患者内皮祖细胞的影响

Effects of vitamin D on endothelial progenitor cells from patients with active systemic lupus erythematosus

目的:观察活动期系统性红斑狼疮(SLE)患者内皮祖细胞(EPCs)数量及功能的变化,并探讨维生素D对SLE患者EPCs数量及功能的影响.方法:收集30例SLEDAI评分大于8分,病情均处于活动期的SLE患者,30例年龄与性别匹配的健康人作为正常对照;酶联免疫吸附试验(ELISA)检测外周血25羟基维生素D[25(OH)D]水平;密度梯度离心和贴壁培养法分离培养EPCs,流式细胞术检测CD34^+/VEGFR-2^+EPCs在全血中的比例;通过计数再贴壁和构建侵袭小室检测EPCs黏附和迁移能力.结果:(1)体外培养过程中,活动期SLE患者EPCs数量(0.028±0.017)%显著低于正常对照组(0.067±0.012)%,有统计学差异(P<0.05);(2)活动期SLE患者EPCs迁移率(1.7±0.9)‰及黏附能力(19±7)显著低于正常对照组EPCs迁移率(3.1±1.6)‰及黏附能力(34±11),有统计学差异(P<0.05);(3)活动期SLE患者外周血25(OH)D质量浓度(14.47±10.39)ng/mL水平低于健康对照组质量浓度(24.15±7.98)ng/mL,有统计学差异(P<0.05);(4)25(OH)D能够增加活动期SLE患者EPCs数量(0.045±0.012)%,高于未加25(OH)D组(0.031±0.012)%,有统计学差异(P<0.05);(5)25(OH)D能够增加活动期SLE患者EPCs迁移率(2.6±0.7)‰及黏附能力(24±9),高于未加25(OH)D组迁移率(1.3±0.8)‰及黏附能力(13±6),有统计学差异(P<0.05).结论:SLE患者维生素D水平降低,可诱发Ⅰ型干扰素通路活化,从而导致EPCs数量和/或功能异常,最终造成狼疮患者血管内皮损伤后修复障碍引发动脉粥样硬化.

Objective: To investigate the effects of vitamin D on Endothelial Progenitor Cells from patients with active systemic lupus erythematosus. Methods: Thirty consecutive patients( SLEDAI score≥8) in the active phase of SLE,also in-patients of our Hospital,aged 30 years,were included in the study,and a group of sex-matched healthy people was used as a control. Enyme linked immune sorbent assay( ELISA) was used to determine the level of 25-dihydroxyvitamin D[25( OH) D] in the peripheral blood. Endothelial progenitor cells( EPCs) were isolated by density gradient centrifugation and cultured.CD34^+/VEGFR-2^+EPCs ratios were analyzed by Flow cytometry. Migration and adhesion of EPCs wereobserved by transwell migration assay. Statistical analysis was conducted with t-test and Mann-Whitney rank test. Results:(1) In active SLE subgroups,the number of EPCs( 0. 028 ± 0. 017) % were significantly lower than the normal control( 0. 067 ± 0. 012) %( P < 0. 05).(2) The migration rate of EPCs from the active SLE patients( 1. 7 ± 0. 9) ‰ was significantly reduced as compared to the normal control( 3. 1 ± 1. 6) ‰( P < 0. 05). Adhesion of EPCs from the active SLE patients( 19 ± 7) was declined as compared to the normal control( 34 ± 11)( P < 0. 05).(3) The level of peripheral blood 25( OH) D in patients with active SLE( 14. 47 ± 10. 39) ng/ml was lower than the normal control( 24. 15 ±7. 98) ng/mL( P < 0. 05).(4) 25( OH) D caused a significant increase in cell number( 0. 045 ±0. 012) % of EPCs from the active SLE patients compared to the untreated cells( 0. 031 ± 0. 012) %( P < 0. 05).(5)25( OH) D caused a significant increase in cell migration( 2. 6 ± 0. 7) ‰ and adhesion( 24 ± 9) of EPCs from the active SLE patients compared to those of untreated cells,which were( 1. 3 ±0. 8) ‰ and( 13 ± 6),respectively( P < 0. 05). Conclusion: Both number and function of EPCs were significantly decreased in SLE patients. Vitamin D may be involved in reduction and dysfunction of EPCs in SLE.