Objective To explore whether macrophage migration inhibitor factor (MIF)can protect human bone microvascular endothelial cells (HBMECs)from glucocorticoid-induced damage.Methods HUVECs were isolated from human femoral head.After HUVECs were cultured and identified,we constructed the ECs damage model with high-dose hydrocortisone.The cells were randomly divided into blank control group,low-dose MIF group,high-dose MIF group with corresponding treatment.Cell activity was detected by AlamarBlue in each group. The number of viable cells was detected in Live/Dead staining.The cell morphology was observed after cytoskeleton staining.Cell migration ability was compared by scratch test and the level of VEGF expression was detected by ELISA.Results Cell model was successfully constructed.The activity of cells in high-dose MIF group (178.3± 15.2)% was significantly higher than that in the control group (100±8.4)% and low-dose MIF group (149.1± 13.8)% (P<0.05).The number of viable cells in high-dose MIF group (139.5±14.3)% was higher than that in low-dose MIF group (121.3±12.9)% while the two groups had more viable cells than the control group (100± 8.4)% (P<0.05).The scratch test results indicated that cell migration ability in high-dose group was the strongest and the scratch disappeared at 24 hours after scratching.The expression of VEGF at 24 hours after intervention was (170±15.7)pg/mL in normal group,(328±25.3)pg/mL in low-dose group and (405±31.2)pg/mL in high-dose group.VEGF level was lower in low-dose group than in high-dose group (P<0.05),but higher than the normal group (P<0.01).Conclusion MIF can promote the proliferation and migration of ECs in a dose-dependent manner and upregulate the expression of VEGF.MIF can improve ECs damage induced by high-dose glucocorticoid.%目的 探讨巨噬细胞移动抑制因子(MIF)对糖皮质激素(Glc)诱导的股骨头内微血管内皮细胞(HUVECs)损伤的治疗作用.方法 从人股骨头标本分离、培养以及鉴定HUVECs,构建Glc诱导的内皮细胞损伤模型,分为空白对照组、低剂量MIF组、高剂量MIF组,予以对应干预.AlamarBlue检测评估细胞活性,Live/Dead细胞染色检测活细胞数目,细胞骨架染色观察各组细胞形态,划痕实验对比各组细胞的迁徙能力,ELISA方法检测细胞分泌VEGF的水平.结果 细胞模型构建成功.高剂量MIF组细胞活性百分比为(178.3±15.2)%,显著高于空白对照组(100±8.4)%及低剂量MIF组(149.1±13.8)%(P<0.05).高剂量MIF组相对活细胞数量(139.5±14.3)%高于低剂量MIF组(121.3±12.9)%,两组均高于空白对照组(100±8.4)%(P<0.05).空白对照组细胞骨架形态异常,MIF干预组细胞骨架形态正常.划痕实验结果提示,高剂量MIF组细胞迁移能力最强,划线后24 h划痕消失.干预24 h空白对照组VEGF表达量为(170±15.7)pg/mL,低剂量MIF组VEGF表达量为(328±25.3)pg/mL,高剂量MIF组VEGF表达量为(405±31.2)pg/mL.低剂量组的VEGF表达水平低于高剂量组(P<0.05),但高于空白对照组(P<0.01).结论 MIF具有促进HUVECs增殖、迁徙作用,具有剂量依赖关系,同时可以上调VEGF的表达,可以改善Glc诱导的受损内皮细胞的形态和功能.
展开▼
