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| 过表达HIF-1αmRNA的小鼠BMSCs经大黄素干预后成骨分化效果实验研究 |
| Effect of emodin on osteogenic differentiation by mouse BMSCs with over-expressing HIF-1α mRNA |
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| DOI：10.3969/j.issn.1006-7108.2026.09.011 |
| 中文关键词: 大黄素 低氧诱导因子-1α 骨髓间充质干细胞 成骨细胞 骨质疏松 |
| 英文关键词:emodin hypoxia inducible factor-1α bone marrow mesenchymal stem cells osteoblast osteoporosis |
| 基金项目:新疆维吾尔自治区卫生健康青年医学科技人才专项（WJWY-202332）；新疆医科大学第六附属医院专项-自然科学基金（LFYKYZX2023-07） |
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| 中文摘要: |
| 目的 探讨大黄素（emodin，ED）对慢病毒介导的低氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）过表达的小鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）向成骨细胞分化的影响。方法 体外传代培养空载慢病毒（HIF-1α-NC）和过表达慢病毒（HIF-1α-OE）感染的小鼠BMSCs并设置分组如下：A组为BMSCs（HIF-1α-NC）+成骨培养基，B组为BMSCs（HIF-1α-OE）+成骨培养基，C组为BMSCs（HIF-1α-NC）+成骨培养基+ED，D组为BMSCs（HIF-1α-OE）+成骨培养基+ED，培养7 d后进行细胞增殖率检测；ELISA检测各组细胞中碱性磷酸酶（alkaline phosphatase，ALP）；qPCR检测ALP、Runt相关转录因子2（runt related transcription factor 2，Runx2）；Western blot检测骨钙素（osteocalcin，OCN）、骨桥蛋白（osteopontin，OPN）、骨唾液酸蛋白（bone sialoprotein，BSP）、Ⅰ型胶原蛋白α1链（collagen type I alpha 1 chain，COLIA1）等成骨分化标志物。结果 培养于含10μmol/L ED的成骨细胞培养基的HIF-1α过表达小鼠BMSCs（D组）中ALP、OCN、OPN、BSP、COLIA1水平较A、B、C组显著增加（P＜0.05），且各组细胞中Runx2水平变化与成骨标志物呈一致性改变。结论 大黄素促进HIF-1α过表达的小鼠BMSCs成骨分化可能与上调关键转录因子Runx2有关。 |
| 英文摘要: |
| Objective To investigate the effect of emodin (ED) on osteoblast differentiation by mouse bone marrow mesenchymal stem cells (BMSCs) with over-expressing hypoxia inducible factor-1α (HIF-1α) mediated by lentivirus. Methods BMSCs of mice infected with empty lentivirus (HIF-1α-NC) and over-expressed lentivirus (HIF-1α-OE) were subcultured in vitro and grouped as follows: group A (BMSCs with HIF-1α-NC+osteogenic medium), group B (BMSCs with HIF-1α-OE+osteogenic medium, group C (BMSCs with HIF-1α-NC+osteogenic medium+ED), and group D (BMSCs with HIF-1α-OE+osteogenic medium+ED). The cell proliferation rate was detected after 7 days of culture. Alkaline phosphatase (ALP) was detected using ELISA. HIF-1α and Runt-related transcription factor 2 (Runx2) were detected using qPCR. The levels of osteocalcin (OCN), osteopontin (OPN), bone sialoprotein (BSP), type I collagen α1 chain (COLIA1), and other osteoblast differentiation markers were detected with Western blotting. Results The levels of ALP, OCN, OPN, BSP, COLIA1, and other markers of osteogenic differentiation in HIF-1α over-expressing mouse BMSCs (Group D) cultured in osteoblast medium rich in 10 μmol/L ED were significantly higher than those in group A/B/C (P<0.05), and the changes of Runx2 levels in cells of each group were consistent with those of osteogenic markers. Conclusion The osteogenic differentiation of mouse BMSCs promoted by emodin through HIF-1α overexpression may be related to the up-regulation of the key transcription factor Runx 2. |
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