巨噬细胞移动抑制因子治疗糖皮质激素性股骨头坏死的细胞学研究

发布时间：2018-07-12 15:49

本文选题：股骨头坏死 + 糖皮质激素　；参考：《西安交通大学学报(医学版)》2017年02期

【摘要】：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)%(P0.05)。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)%(P0.05)。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24h划痕消失。干预24h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(P0.05),但高于空白对照组(P0.01)。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。
[Abstract]:Objective to investigate the therapeutic effect of macrophage migration inhibitor (MIF) on glucocorticoid (Glc) -induced injury of vascular endothelial cells (HUVECs) in femoral head. Methods HUVECs were isolated, cultured and identified from human femoral head specimens. The model of endothelial cell injury induced by Glc was established and divided into three groups: blank control group, low dose MIF group and high dose MIF group. The number of living cells was detected by living / Dead cell staining, the morphology of each group was observed by cytoskeleton staining, the migration ability of each group was compared by scratch test and the level of VEGF secreted by cells was detected by Elisa. Results the cell model was successfully constructed. The percentage of cell activity in the high dose MIF group was (178.3 卤15.2), significantly higher than that in the blank control group (100 卤8.4)% and the low dose MIF group (149.1 卤13.8)% (P0.05). The number of relative living cells in the high dose MIF group (139.5 卤14.3)% was higher than that in the low dose MIF group (121.3 卤12.9), and both groups were higher than the blank control group (100 卤8.4)% (P0.05). The morphology of cytoskeleton was normal in MIF intervention group. The results of scratch test showed that the migration ability of high dose MIF group was the strongest, and the scratch disappeared 24 hours after marking. The expression of VEGF was (170 卤15.7) PG / mL in control group, (328 卤25.3) PG / mL in low dose MIF group, and (405 卤31.2) PG / mL in high dose MIF group. The expression of VEGF in low dose group was lower than that in high dose group (P0.05), but higher than that in blank control group (P0.01). Conclusion MIF can promote the proliferation and migration of HUVECs in a dose-dependent manner. MIF can up-regulate the expression of VEGF and improve the morphology and function of injured endothelial cells induced by Glc.
【作者单位】：西安交通大学第二附属医院骨一科;
【基金】：国家自然科学基金资助项目(No.81301562)~~
【分类号】：R681.8

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