Department of Joint Surgery, Osteonecrosis and Joint Reconstruction Ward, Honghui Hospital, Xi'an Jiaotong University, China.
Beijing Key Laboratory of Arthritic and Rheumatic Diseases, China-Japan Friendship Hospital, China.
Adv Clin Exp Med. 2020 Mar;29(3):345-353. doi: 10.17219/acem/112602.
The pathogenesis of glucocorticoid (GC)-induced osteonecrosis (ON) of the femoral head remains unclear. Recent research has suggested that it is closely associated with injured bone microvascular endothelial cells (BMECs). However, few studies have used BMECs to perform research pertaining ON of the femoral head.
The objective of this study was to investigate the functional changes of BMECs treated with a GC and to detect the changes in related genes using microarrays.
Cells were isolated using an enzymatic method and identified with EC markers, such as von Willebrand factor (vWF), CD31 and vascular endothelial cadherin (VE-cadherin). Bone microvascular endothelial cells were treated with 0.1 mg/mL and 0.3 mg/mL of hydrocortisone to establish a GC-damaged model of BMECs. The mRNA microarrays were used to detect the differential expression profiles between BMECs with and without GC damage.
Primary cells appeared as having a cobblestone-like morphology. Immunofluorescence staining revealed that the cells were 100% positive for vWF and CD31, and near 100% positive for VE-cadherin. It also confirmed that the cells were BMECs. Bone microvascular endothelial cells treated with 0.1 mg/mL of hydrocortisone showed shrinkage, and those treated with 0.3 mg/mL of hydrocortisone mostly showed apoptosis. The mRNA microarray showed that genes associated with endothelial cells, such as endothelin 1 (ET-1) receptor, angiotensin II (AII) receptor, intercellular adhesion molecule 1 (ICAM-1), and plasminogen activator inhibitor 1 (PAI-1), were upregulated, and genes associated with endothelial nitric oxide synthase (eNOS), endothelin 1 (ET-1), prostaglandin I2 (PGI2) synthase, PGI2 receptor, vascular endothelial growth factor (VEGF), prostaglandin E (PGE) synthase, and PGE receptor were downregulated. The results of quantitative polymerase chain reaction (qPCR) validation were consistent with the findings of mRNA microarrays.
Glucocorticoids promoted BMECs to express vasoconstrictors and procoagulant factors and related receptors, and decreased the expression of vasodilators and their receptors.
糖皮质激素（GC）诱导的股骨头坏死（ON）的发病机制尚不清楚。最近的研究表明，它与受损的骨微血管内皮细胞（BMECs）密切相关。然而，很少有研究使用 BMECs 来研究股骨头坏死。
本研究旨在探讨 GC 处理的 BMECs 的功能变化，并通过微阵列检测相关基因的变化。
采用酶法分离细胞，并用血管性血友病因子（vWF）、CD31 和血管内皮钙黏蛋白（VE-cadherin）等 EC 标志物鉴定。用 0.1mg/ml 和 0.3mg/ml 的氢化可的松处理骨微血管内皮细胞，建立 BMECs 的 GC 损伤模型。采用 mRNA 微阵列检测 BMECs 有无 GC 损伤的差异表达谱。
原代细胞呈鹅卵石样形态。免疫荧光染色显示，细胞 vWF 和 CD31 均为 100%阳性，VE-cadherin 近 100%阳性。证实细胞为 BMECs。用 0.1mg/ml 氢化可的松处理的骨微血管内皮细胞出现收缩，用 0.3mg/ml 氢化可的松处理的细胞大多出现凋亡。mRNA 微阵列显示，与内皮细胞相关的基因，如内皮素 1（ET-1）受体、血管紧张素 II（AII）受体、细胞间黏附分子 1（ICAM-1）和纤溶酶原激活物抑制剂 1（PAI-1）上调，与内皮型一氧化氮合酶（eNOS）、内皮素 1（ET-1）、前列环素 I2（PGI2）合酶、PGI2 受体、血管内皮生长因子（VEGF）、前列腺素 E（PGE）合酶和 PGE 受体下调。定量聚合酶链反应（qPCR）验证结果与 mRNA 微阵列一致。
糖皮质激素促进 BMECs 表达血管收缩剂和促凝因子及其相关受体，减少血管扩张剂及其受体的表达。
