1. 甘肃中医药大学,兰州 730000;2. 甘肃中医药大学附属医院,兰州 730000;3. 成都中医药大学, 成都 610000;4. 宝鸡市中医医院,陕西 宝鸡 721000
1. Gansu University of Chinese Medicine, Lanzhou 730000, China. 2. Affiliated Hospital of Gansu University of Chinese Medicine, Lanzhou 730000. 3. Chengdu University of Chinese Medicine, Chengdu 610000. 4. Baoji Hospital of Chinese Medicine, Baoji 721000
目的 观察 Hif-1α/ VEGF 信号轴和骨中特异性 H 型血管在 SINFH 模型大鼠股骨头中的改变,探究其在骨坏死发生中的改变及意义。 方法 30 只 SPF 级雄性 SD 大鼠随机分为空白组(CG)、模型组(MG)、去铁胺 组(DFO),每组 10 只。 MG 组及 DFO 组采用甲强龙联合脂多糖法行 SINFH 造模,CG 组及 MG 组腹腔注射生理盐水,DFO 组予腹腔注射 250 mg / (kg·d)甲磺酸去铁胺。 造模 6 周后,运用 Micro-CT 分析观察股骨头微观结构变化,HE 染色观察股骨头组织病理变化,免疫荧光染色分析股骨头中 H 型血管改变,RT-PCR 分析股骨头中 Hif-1α/ VEGF 信号轴相关因子的表达。 结果 Micro-CT 分析显示,MG 组、DFO 组股骨头均出现骨小梁稀疏,但 MG 组骨小梁出现断裂情况,且股骨头软骨下囊性变形成。 HE 染色显示,与 CG 组比,MG 组和 DFO 组均出现明显的股骨头 坏死(P< 0. 01)。 免疫荧光染色显示,与 CG 组比较,MG 组股骨头中 H 型血管量显著下降(P< 0. 05),与 MG 组比 较 DFO 组股骨头中 H 型血管量显著增加(P< 0. 01)。 与 CG 组比,MG 组 Osterix+ 成骨(祖) 细胞量均出现下降 (P< 0. 01),与 MG 组比,DFO 组 Osterix+成骨(祖)细胞量增加(P< 0. 01)。 RT-PCR 分析显示,与 CG 比较 MG 组 大鼠股骨头 Hif-1α、VEGF、Osterix、Runx2(mRNA)表达下降(P< 0. 01),与 MG 组比,DFO 组 Hif-1α、VEGF、Osterix、 Runx2(mRNA)表达升高(P< 0. 01)。 结论 在 SINFH 大鼠模型中,激素诱导 Hif-1α/ VEGF 信号轴调控障碍,以及特异性 H 型血管发生损害,提示骨特异的 H 型血管损害可能是激素性股骨头坏死的关键发病机制之一。
Objective To observe the changes in the hypoxia-inducible factor 1α (Hif-1α) / vascular endothelial growth factor (VEGF) signaling axis and specific type H vessels in the femoral head of steroid-induced osteonecrosis of the femoral head ( SONFH) model rats and explore the role of these changes in the development of osteonecrosis. Methods Thirty specific pathogen-free grade male Sprague-Dawley rats were randomly divided into a control group ( CG), model group ( MG) and deferoxamine group ( DFOG), with 10 rats in each group. The MG and DFOG were treated with methylprednisolone combined with lipopolysaccharide for SONFH modeling, the CG and MG received intraperitoneal injections of normal saline, and the DFOG received intraperitoneal injections of 250 mg / kg deferoxamine mesylate. Six weeks after modeling, micro-computed tomography was performed to observe the microstructural changes of the femoral head, hematoxylin / eosin staining was used to observe the histopathological changes of the femoral head, immunofluorescence staining was used to analyze the changes of type H vessels in the femoral head, and reverse- transcription polymerase chain reaction (RT-PCR) was used to analyze expression of HIF-1α/ VEGF signaling axis-related factors in the femoral head. Results Micro-computed tomography analysis showed that the femoral head in the MG and DFOG had sparse bone trabeculae, but the bone trabeculae in the MG exhibited fractures and the femoral head showed subchondral cystic degeneration. Hematoxylin / eosin staining showed that compared with the CG, both the MG and DFOG had obvious femoral head necrosis (P< 0. 01). Immunofluorescence staining showed that compared with the CG, the amount of type H vessels in the femoral head was significantly lower in the MG (P< 0. 05) and significantly higher in the DFOG (P< 0. 01). Compared with the CG, the amount of Osterix+ osteoblast (progenitor) cells was significantly lower in the MG (P< 0. 01) and significantly higher in the DFOG (P< 0. 01). RT-PCR showed that the expression of HIF-1α, VEGF, osterix, and Runx2 (mRNA) in the femoral head was significantly lower in the MG than CG (P< 0. 01) and significantly higher in the DFOG than MG ( P< 0. 01). Conclusions Steroid-induced HIF-1α/ VEGF signaling axis dysregulation and specific type H vessel damage were observed in this SONFH rat model, suggesting that bone-specific type H vessel damage may be a key pathogenetic factor of SONFH.
于海洋,卢增鹏,汪海燕,曹盼举,姚川江,曹林忠,田杰祥,张晓刚.激素性股骨头坏死中 Hif-1α / VEGF 信号轴和 H 型血管改变的实验研究[J].中国实验动物学报,2022,30(6):759~766.复制