糖皮质激素对人股骨头骨微血管内皮细胞功能的影响

Effect of glucocorticoid on the function of bone microvascular endothelial cells derived from human femoral head

[目的]通过基因芯片对人股骨头骨微血管内皮细胞表达谱分析及荧光定量PCR验证,探讨糖皮质激素对骨微血管内皮细胞损伤后发生的功能变化。[方法]采用本实验室建立的方法进行人股骨头骨微血管内皮细胞的分离、培养及鉴定,证明细胞为骨微血管内皮细胞。用0.1 mg/ml的含氢化可的松的培养基培养细胞,建立糖皮质激素骨微血管内皮细胞损伤模型。设正常对照组,用晶芯mRNA表达谱芯片对细胞损伤模型和对照组进行差异性转录本检测,对差异性表达基因行q PCR验证。[结果]mRNA芯片表达基因筛选结果发现ICAM-1、ET-1受体、PAI-1、血管紧张素II受体较对照组明显上调,e NOS、ET-1、PGI2合成酶、VEGF、PGE合成酶及PGE受体表达明显下调。q PCR验证结果与芯片结果一致。[结论]糖皮质激素促进了人股骨头骨微血管内皮细胞分泌缩血管因子、促凝血的因子及相关受体的表达,降低了舒张血管因子及相应受体的表达。

[Objective] To investigate the influence of glucocorticoid on gene expression of bone microvascular endothelial cells（ BMECs） derived from human femoral head using microarray analysis and q PCR in vitro. [Methods] BMECs were isolated from adult bone marrow inside femoral head according to our previously established method. Then,the BMECs were cultured and identified for following experiments. BMECs were treated by the hydrocortisone with concentration of 0. 1ml / ml in endothelial cell medium. The control group was cultured without hydrocortisone. The difference of gene expression profile in those group was detected by the mRNA microarrays. Furthermore,some specific genes were identified and validated by q PCR. [Results] With the hydrocortisone,the expression of endothelin 1 receptor,angiotensin II receptor,intercellular adhesion molecule 1 and plasminogen activator inhibitor 1 was significantly upregulated. Meanwhile,the expression of nitric oxide synthase,endothelin 1,prostaglandin I2 synthase,prostaglandin I2 receptor,vascular endothelial growth factor,prostaglandin E synthase and prostaglandin E receptor was downregulated. The results of mRNA microarrays were further validated by q PCR. [Conclusion] Glucocorticoids may upregulate the gene expression of vasoconstrictors,procoagulant factors and related receptor,and meanwhile downregulate the gene expression of vasodilators and the corresponding receptors.