基质细胞衍生因子-1α对牛视网膜血管内皮细胞CXCR-4和VEGF表达的影响

Effects of stromal cell-derived factor-1α on expression of CXCR-4 and VEGF in bovine retinal vascular endothelial cells

目的观察基质细胞衍生因子-1α(stromal cell-derived factor-1α,SDF-1α)对体外培养的牛视网膜血管内皮细胞(bo-vine retinal vascular endothelial cells,BRVEC)表达趋化性细胞因子受体-4(chemotaxis cytokine receptor-4,CXCR-4)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的影响,探讨SDF-1α对BRVEC作用的机制。方法取第3-7代选择性培养的BRVEC悬液接种于12孔培养板,常规培养条件下分别用不同浓度的糖和不同浓度的SDF-1α及糖+SDF-1α干预,收集上清液ELISA检测VEGF的表达,收集细胞Western blotting检测CXCR-4的表达。结果正常浓度葡萄糖(5.6 mmol.L-1)对BRVEC表达VEGF和CXCR-4均无影响,高糖(25.0 mmol.L-1)增强BRVEC表达CXCR-4和VEGF,并呈时间依赖性。SDF-1α促进BRVEC表达VEGF和CXCR-4,并呈剂量和时间依赖性,SDF-1α的作用在100μg.L-1达高峰,CXCR-4表达增加水平较VEGF明显。高糖联合SDF-1α培养时VEGF和CXCR-4的表达在48 h达高峰,分别为(230.17±19.74)ng.L-1、(0.96±0.12)ng.L-1,均显著高于SDF-1α单独培养时,差异均有统计学意义(P均<0.05),高糖+SDF-1α干预时,二者协同促进BRVEC表达VEGF和CXCR-4,且CXCR-4表达增加水平较VEGF明显。结论高糖和SDF-1α均能刺激BRVEC表达CXCR-4和VEGF,并且具有协同作用。

Objective To evaluate the effects and mechanisms of stromal cell-derived factor-1α（SDF-1α） on the expression of chemotaxis cytokine receptor-4（CXCR-4） and vascular endothelial growth factor（VEGF） in bovine retinal microvascular endothelial cells（BRVEC） in vitro.Methods The suspension of the 3rd to the 7th generation BRVEC were inoculated into 12 holes plate.Under common cultural condition,BRVEC were interfered with glucose,SDF-1α and glucose combined with SDF-1α at different concentrations.Supernatant was obtained to detect the level of VEGF by ELISA and cells were obtained to detect the expression of CXCR-4 by Western blotting.Results Normal glucose（5.6 mmol·L-1） did not affect the expression of CXCR-4 and VEGF.High glucose（25.0 mmol·L-1） stimulated the expression of CXCR-4 and VEGF in BRVEC at a time-dependent manner.SDF-1α promoted the expression of VEGF and CXCR-4 in BRVEC at a dose-and time-dependent manner,and its effect achieved peak at the concentration of 100 μg·L-1,the level of CXCR-4 increased obviously higher than VEGF did.The expression of VEGF and CXCR-4 by the combining culture of high glucose and SDF-1α went to peak at 48 hours,were（230.17±19.74）ng·L-1 and（0.96±0.12）ng·L-1,respectively,and both were significantly higher than that culture with SDF-1α alone,the difference was statistically significant（all P〈0.05）.Both high glucose and SDF-1α could stimulate the expression of VEGF and CXCR-4 in BRVEC at a synergetic manner,and the level of CXCR-4 increased more obvious than VEGF did.Conclusion Both high glucose and SDF-1α can stimulate the expression of CXCR-4 and VEGF in BRVEC at a synergetic manner.