| 何嘉怡,迟瑞敏,贺毅,等.淫羊藿苷通过激活转录共激活因子预防大鼠激素性股骨头坏死进展的实验研究.骨科,2022,13(3): 255-263. |
| 淫羊藿苷通过激活转录共激活因子预防大鼠激素性股骨头坏死进展的实验研究 |
| Icariin Protects against Glucocorticoid-Induced Osteonecrosis of Femoral Head in Rats via Activating TAZ Expression |
| 投稿时间：2022-01-17 |
| DOI：10.3969/j.issn.1674-8573.2022.03.012 |
| 中文关键词: 淫羊藿苷 糖皮质激素 股骨头坏死 转录共激活因子 血管生成 |
| 英文关键词: Icariin Glucocorticoids Osteonecrosis of the femoral head TAZ Angiogenesis |
| 基金项目:国家自然科学基金（82172435） |
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| 中文摘要: |
| 目的 研究淫羊藿苷对激素性股骨头坏死的保护作用及机制。方法 分离SD大鼠骨髓间充质干细胞（rBMSCs）并按以下方式干预：对照组、地塞米松组（1×10-6 mol/L），联合干预组［地塞米松（1×10-6 mol/L）+不同浓度淫羊藿苷（1×10-8 mol/L，1×10-7 mol/L，1×10-6 mol/L，1×10-5 mol/L）］；检测各组细胞增殖、凋亡、成骨/成脂分化情况，以及转录共激活因子（transcriptional co-activator with PDZ-binding motif，TAZ）、结缔组织生长因子（connective tissue growth factor，CTGF）、Runt相关转录因子2（Runt-related transcriptional factor 2，Runx2）蛋白表达变化；siRNA沉默TAZ后，观察各组下游CTGF、Runx2表达变化。体内实验使用52只SD大鼠，随机分为对照组、甲强龙组（甲基强的松龙40 mg·kg-1·d-1）和联合干预组（甲基强的松龙40 mg·kg-1·d-1+淫羊藿苷100 mg·kg-1·d-1）。HE染色评估股骨头骨小梁结构；免疫组化染色评估抗骨钙蛋白（osteocalcin，OCN）、TAZ、血小板-内皮细胞粘附分子（Platelet endothelial cell adhesion molecule-1，PECAM-1/CD31）和血管内皮生长因子（vascular endothelial growth factor，VEGF）表达；TUNEL染色评估细胞凋亡情况；Micro-CT评估股骨头骨密度、骨体积/总体积、骨小梁数、骨小梁间距和骨小梁厚度；血管造影评估股骨头血供情况。结果 体外实验表明地塞米松能促进rBMSCs凋亡和成脂分化，抑制其增殖和成骨分化；而淫羊藿苷能显著削弱这些效应。此外，淫羊藿苷能显著逆转地塞米松对rBMSCs中TAZ、CTGF蛋白表达的影响。抑制TAZ表达后，联合干预组rBMSCs中Runx2的表达也受到抑制。体内实验结果证实淫羊藿苷可保护大鼠股骨头软骨下骨质和股骨头内血液供应免受激素的破坏。激素能显著抑制大鼠股骨头TAZ表达，而淫羊藿苷能促进TAZ表达。结论 淫羊藿苷可通过激活TAZ表达来预防大鼠激素性股骨头坏死的发生发展。 |
| 英文摘要: |
| Objective To investigate the protective effect of icariin on glucocorticoid-induced osteonecrosis of the femoral head (GIOFH) and its underlying mechanism. Methods The rat bone marrow mesenchymal stem cells (rBMSCs) from 3-month-old SD rats were used in vitro study. We set up the following groups: control group; dexamethasone group: dexamethasone (10-6 mol/L); dexamethasone + icariin group: dexamethasone (10-6 mol/L) + icariin (1×10-8 mol/L, 1×10-7 mol/L, 1×10-6 mol/L, 1×10-5 mol/L) at different concentrations. The proliferation, apoptosis, osteogenic differentiation and adipogenic differentiation of each group were detected respectively. Western blot was used to detect the expression changes in transcriptional coactivator with PDZ-binding motif (TAZ), connective tissue growth factor (CTGF) and Runt-related transcriptional factor 2 (Runx2) in each group. Small interference RNA (SiRNA) was used to knock down TAZ and the expression changes of CTGF and Runx2 downstream was detected. In vivo study, 52 SD rats were randomly divided into control group (n=12), methylprednisolone group (n=20) and methylprednisolone + icariin group (n=20). After corresponding treatment, bone trabecular structure was evaluated by H&E staining, and the expression of OCN, TAZ, CD31 and VEGF was evaluated by immunohistochemical staining. TUNEL staining was used to evaluate cell apoptosis. Bone mineral density, bone volume/tissue volume, trabecular number, trabecular spacing, and trabecular thickness of the femoral head were assessed using micro-CT. Blood supply to the femoral head was assessed using angiography. Results The in vitro study showed that dexamethasone could promote apoptosis and inhibit proliferation of rBMSCs, and promote adipogenic differentiation and inhibit osteogenic differentiation. Icariin could significantly reverse these effects. In addition, icariin could significantly reverse the effects of dexamethasone on the expression of TAZ and CTGF proteins in rBMSCs. After inhibition of TAZ expression, the effect of icariin and dexamethasone on Runx2 expression of rBMSCs was eliminated. The in vivo study showed that icariin could protect the subchondral trabecular bone and blood supply from methylprednisolone damage in rats. Methylprednisolone could inhibit the expression of TAZ in rat femoral head, while icariin could promote the expression of TAZ. Conclusion Icariin might inhibit the development of GIOFH in rats by activating TAZ expression. |
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