南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (4): 712-719.doi: 10.12122/j.issn.1673-4254.2024.04.13

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α2-巨球蛋白通过调控血管内皮细胞改善小鼠激素性股骨头坏死

朱 奇，路云翔，彭 优，何嘉乐，韦泽宇，李智勇，陈郁鲜   

  1. 中山大学附属第三医院关节外科创伤骨科，广东 广州 510630
  • 发布日期:2024-04-29

α2-macroglobulin alleviates glucocorticoid-induced avascular necrosis of the femoral head in mice by promoting proliferation, migration and angiogenesis of vascular endothelial cells

ZHU Qi, LU Yunxiang, PENG You, HE Jiale, WEI Zeyu, LI Zhiyong, CHEN Yuxian   

  1. Department of Joint Surgery, Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510630, China
  • Published:2024-04-29

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摘要： 目的 探讨α2-巨球蛋白（A2M）是否对激素性股骨头坏死（SANFH）具有保护作用。方法 体外实验：用梯度浓度（10-8~10-5 mol/L）地塞米松（DEX）处理人脐静脉内皮细胞（HUVECs）建立糖皮质激素（GC）诱导内皮细胞损伤体外模型，设置对照组、DEX组、DEX+A2M（0.05 mg/mL）和 DEX+A2M（0.1 mg/mL）4组，采用CCK-8法检测细胞活性，Transwell实验和划痕愈合实验检测HUVECs迁移，血管形成实验检测HUVECs血管形成能力，Western blot检测HUVECs中CD31和VEGF-A蛋白表达水平。体内实验：将24只BALB/c小鼠分为对照组、模型组（GC）和干预组（GC+A2M），Micro-CT检测骨小梁情况，HE染色观察组织学特征，免疫组化染色检测CD31的表达。结果 与对照组相比，DEX处理后，HUVECs的活力下降，细胞迁移速度减慢，血管形成能力下降；并且DEX处理组细胞活性随着DEX浓度增加和刺激时间延长而降低（P<0.05）；加入A2M后，经DEX处理的HUVECs活性、细胞迁移能力和血管形成能力得以恢复，并和A2M的浓度成正相关（P<0.05）；Western blot结果显示，与对照组相比，DEX降低了HUVECs的CD31和VEGF-A蛋白表达水平，而A2M的介入使经DEX处理的HUVECs的CD31和VEGF-A蛋白表达水平得以恢复（P<0.05）。与对照组相比，GC组小鼠股骨头骨小梁明显破坏，破坏的骨小梁里充斥着大量空骨陷窝和肥大的脂肪细胞，CD31表达下降（P<0.05）；与GC组相比，GC+A2M组骨小梁的破坏较轻，大部分正常骨组织结构存在，并上调CD31的表达（P<0.05）。结论 A2M上调经DEX处理的HUVECs的增殖、迁移和血管形成A2M通过减轻股骨头内微循环损伤及维持微循环的稳定对激素性股骨头坏死起一定的保护作用。

关键词: α2-巨球蛋白；激素性股骨头坏死；内皮功能障碍；血管内皮生长因子-A

Abstract: Objective To explore the mechanism underlying the protective effect of α2-macroglobulin (A2M) against glucocorticoid-induced femoral head necrosis. Methods In a human umbilical vein endothelial cell (HUVEC) model with injuries induced by gradient concentrations of dexamethasone (DEX; 10-8 -10-5 mol/L), the protective effects of A2M at 0.05 and 0.1 mg/mL were assessed by examining the changes in cell viability, migration, and capacity of angiogenesis using CCK-8 assay, Transwell and scratch healing assays and angiogenesis assay. The expressions of CD31 and VEGF-A proteins in the treated cells were detected using Western blotting. In BALB/c mouse models of avascular necrosis of the femoral head induced by intramuscular injections of methylprednisolone, the effects of intervention with A2M on femoral trabecular structure, histopathological characteristics, and CD31 expression were examined with Micro-CT, HE staining and immunohistochemical staining. Results In cultured HUVECs, DEX treatment significantly reduced cell viability, migration and angiogenic ability in a concentration- and time-dependent manner (P<0.05), and these changes were obviously reversed by treatment with A2M in positive correlation with A2M concentration (P<0.05). DEX significantly reduced the expression of CD31 and VEGF-A proteins in HUVECs, while treatment with A2M restored CD31 and VEGF-A expressions in the cells (P<0.05). The mouse models of femoral head necrosis showed obvious trabecular damages in the femoral head, where a large number of empty lacunae and hypertrophic fat cells could be seen and CD31 expression was significantly decreased (P<0.05). A2M treatment of the mouse models significantly improved trabecular damages, maintained normal bone tissue structures, and increased CD31 expression in the femoral head (P<0.05). Conclusion A2M promotes proliferation, migration, and angiogenesis of DEX-treated HUVECs and alleviates methylprednisolone-induced femoral head necrosis by improving microcirculation damages and maintaining microcirculation stability in the femoral head.

Key words: α2-macroglobulin; steroid-induced avascular necrosis of the femoral head; endothelial dysfunction; vascular endothelial growth factor-A