Gao F Q, Han J, Zhang Q Y, Ma J H, Sun W, Cheng L M, Li Z R, Ma J
Osteonecrosis and Joint Preservation Reconstruction Center, Beijing Key Laboratory of Immune Inflammatory Diseases, Department of Orthopedics, China-Japan Friendship Hospital, Beijing 100029, China.
Department of Orthopedics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, China.
Zhonghua Yi Xue Za Zhi. 2020 Nov 24;100(43):3457-3462. doi: 10.3760/cma.j.cn112137-20200331-01029.
To investigate the expression levels and activation differences of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) gene in bone microvascular endothelial cells (BMECs) in different regions of human femoral head. Tissue specimens of femoral heads were obtained from hip arthroplasty carried out in China-Japan Friendship Hospital from January 2017 to June 2018. And the BMECs we isolated, purified, identified and cultured from different regions of the human femoral head: in the subchondral and cancellous bone regions. The BMECs from the two regions were intervened by hydrocortisone with a series of low concentration gradients (0, 0.03, 0.06, 0.10 mg/ml) respectively. The cell phenotype and functional status of BMECs and cell migration were detected by scratch experiments, and the angiogenesis in different regions of the femoral head was observed. The mRNA and protein expression of 11beta-HSD1, 11beta-HSD2 in BMECs were detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) and Western-blot method, respectively. With the increase of the concentration of hydrocortisone, the 11beta-HSD1 mRNA and protein expression of BMECs in the subchondral and cancellous bone regions of the femoral head increased significantly, and the 11beta-HSD1 mRNA and protein expression of BMECs in the subchondral bone region was significantly lower than those in cancellous bone region (all 0.05). The 11beta-HSD2 mRNA and protein expression of BMECs in the cancellous bone region showed a slow decrease first and then increased slightly at 0.10 mg/ml, while the expression in the subchondral bone region was the opposite. The 11beta-HSD2 mRNA and protein expression of BMECs in subchondral bone region was slightly higher than those in cancellous bone region (all 0.05), but there was no significant statistical difference between the two regions at 0.10 mg/ml (0.123±0.018 vs 0.126±0.021, 0.577±0.231 vs 0.609±0.174, 1.380, 0.409, both 0.05). At different times of the 0.06 mg/ml hydrocortisone intervention, there was no significant differences in scratch closure rate, the number of BMECs lumen, the number of buds and the length of tubule branches in different regions of the femoral head (all 0.05). The 11beta-HSD expression of BMECs in different regions of human femoral head is significantly different. The 11beta-HSD1 is high-expressed, but 11beta-HSD2 is low-expressed in BMECs of the cancellous bone region, and those are opposite in the subchondral bone region, which helps to explain the pathological characteristics and pathogenesis of hormonal osteonecrosis.
探讨人股骨头不同区域骨微血管内皮细胞（BMECs）中11β-羟基类固醇脱氢酶（11β-HSD）基因的表达水平及激活差异。收集2017年1月至2018年6月在中日友好医院行髋关节置换术患者的股骨头组织标本。并从人股骨头的不同区域（软骨下骨区域和松质骨区域）分离、纯化、鉴定和培养BMECs。分别用一系列低浓度梯度（0、0.03、0.06、0.10 mg/ml）的氢化可的松干预两个区域的BMECs。通过划痕实验检测BMECs的细胞表型、功能状态及细胞迁移情况，并观察股骨头不同区域的血管生成情况。分别采用实时荧光定量聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测BMECs中11β-HSD1、11β-HSD2的mRNA和蛋白表达。随着氢化可的松浓度的增加，股骨头软骨下骨区域和松质骨区域BMECs的11β-HSD1 mRNA和蛋白表达均显著增加，且软骨下骨区域BMECs的11β-HSD1 mRNA和蛋白表达显著低于松质骨区域（均P<0.05）。松质骨区域BMECs的11β-HSD2 mRNA和蛋白表达先缓慢下降，在0.10 mg/ml时略有升高，而软骨下骨区域的表达情况则相反。软骨下骨区域BMECs的11β-HSD2 mRNA和蛋白表达略高于松质骨区域（均P<0.05），但在0.10 mg/ml时两个区域之间无显著统计学差异（0.123±0.018 vs 0.126±0.021，0.577±0.231 vs 0.609±0.174，F=1.380，P=0.409，均P>0.05）。在0.06 mg/ml氢化可的松干预的不同时间，股骨头不同区域的划痕愈合率、BMECs管腔数、芽数及小管分支长度均无显著差异（均P>0.05）。人股骨头不同区域BMECs的11β-HSD表达存在显著差异。松质骨区域BMECs中11β-HSD1高表达，而11β-HSD2低表达，在软骨下骨区域则相反，这有助于解释激素性骨坏死的病理特征及发病机制。
