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| 补肾调肝方诱导衰老BMSCs成骨分化的 实验研究 |
| Experimental study of the osteogenic differentiation of aging bone mesenehymal stem cells induced by kidney-tonifying and liver-regulating recipes |
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| DOI： |
| 中文关键词: 原发性骨质疏松症 骨髓间充质干细胞 补肾调肝 |
| 英文关键词:Primary osteoporosis Bone mesenehymal stem cells Kidney-tonifying and liver-regulating recipes |
| 基金项目:广东省社会发展领域科技计划项目（2011028) |
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| 中文摘要: |
| 目的深人研究补肾调肝方诱导衰老骨髓间充质干细胞（Bone Mesenehymal Stem Cells,BMSCs)成骨分化的效应，探讨补肾调肝方抗骨质疏松的机制。方法贴壁培养法分离纯化大鼠 BMSCs,用D-半乳糖诱导制作BMSCs衰老模型，卩-半乳糖苷酶染色检测BMSCs衰老情况。将培养成 功的衰老BMSCs分为空白组、成骨诱导组和中药组，培养7d、l4d后行矿化结节茜素红染色（Alizarin Red S,ARS),倒置显微镜下观察矿化骨结节形成情况。结果D-半乳糖诱导BMSCs呈现典型的细 胞衰老形态和结构变化。在成骨诱导剂、补肾调肝方水提液的作用下，与空白组比较，衰老的BMSCs 钙化结节数量明显增多，差异有非常显著的统计学意义<0.01)。中药组的矿化结节明显多于成 骨诱导组，差异有非常显著的统计学意义（P<0.01)。结论D-半乳糖具有诱导SD大鼠BMSCs衰老的作用，采用D-半乳糖诱导培养可建立BMSCs衰老模型。补肾调肝方抗骨质疏松的机制之一是通 过促进衰老BMSCs成骨分化。 |
| 英文摘要: |
| Objective To investigate the osteogenic differentiation of aging bone mesenehymal stem cells (BMSCs) induced by kidney-tonifying and liver-regulating recipes，and to explore its anti-osteoporosis mechanism. Methods BMSCs of SD rats were isolated with adherent separation method and primarily cultured in vitro. BMSCs senescence model was induced by D-galactose, and the senescence of BMSCs was detected by p-gal dyeing. Successfully cultured aging BMSCs were divided into 3 groups ： blank group, osteo-induced group, and Chinese medicine group. After 7-day and 14-day culturing, Alizarin red staining was performed. And the formation of mineralized nodules was observed under phase-contract microscope.Results Compared with control cells, BMSCs induced by D-galactose displayed morphological and biological senescent phenotype. Using osteogenic inducer and water extract of kidney-tonifying and liver- regulating recipes, the number of mineralized nodules was notably more than that in blank group. The difference was significant ( P < 0. 01 ). The number of mineralized nodules in Chinese medicine group was more than that in osteo-induced group ( P < 0. 01 ). Conclusion D-galactose can induce BMSCs senescence. BMSCs senescence model can be established by culturing with D-galactose. One possible mechanism of kidney-tonifying and liver-regulating recipes in treating osteoporosis is to promote osteogenic differentiation of aging BMSCs. |
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