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| 木通皂苷D促进糖皮质激素环境下小鼠骨髓间充质干细胞成骨分化 |
| Akebia saponin D promotes osteogenic differentiation of bone marrow mesenchymal stem cells under glucocorticoid condition in mice |
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| DOI：10.3969/j.issn.1006.7108.2020.04.012 |
| 中文关键词: 中医中药 糖皮质激素性骨质疏松 木通皂苷D 骨髓间充质干细胞 成骨分化 BMP/Smad信号通路 |
| 英文关键词:Chinese traditional medicine glucocorticoid-induced osteoporosis akebia saponin D bone marrow mesenchymal stem cells osteogenic differentiation BMP/Smad signaling pathway |
| 基金项目:国家自然科学基金（81674000, 81674000, 81674000）；广东省高等学校珠江学者岗位计划资助项目（2018年度）；中华中医药学会2017—2019年度青年人才托举工程项目中华中医药学会立项项目A类（CACM-2017-QNRC1-01）；广东省自然科学基金（2016A030313645）；广东省科学技术厅（2016A020226006）；广州市珠江科技新星（201710010078）；广东省中医药局科研项目（20180330134046, 20161136）；中国博士后科学基金面上资助（2017M610271）；广州中医药大学第一临床学院优秀博士学位论文培育项目（YB201602, YB201702）；广州中医药大学第一附属医院“创新强院”工程科研项目（2017QN08, 2017TD08） |
| 作者 | 单位 | | 张志达1,3 沈耿杨2,3 任辉2,3 余翔1,3 尚奇1,3 黄锦菁1,3 招文华1,3 余佩沅1,3 詹玫琦1,3
梁德2 杨志东2 江晓兵2,3* | 1. 广州中医药大学第一临床医学院，广东 广州 510405
2. 广州中医药大学第一附属医院脊柱骨科，广东 广州 510405
3. 广州中医药大学岭南医学研究中心，广东 广州 510405 |
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| 中文摘要: |
| 目的 观察木通皂苷D（ASD）对糖皮质激素（GC）环境下小鼠骨髓间充质干细胞（BMSCs）成骨分化的影响及其机制。方法 体外培养小鼠BMSCs，实验分为三组，分别用成骨诱导培养基（OBI）、OBI＋地塞米松（DEXA，0.1 μmol/L）、OBI＋DEXA（0.1 μmol/L）＋ASD（10 μmol/L）干预。ALP试剂盒检测ALP活性，茜素红染色检测矿化情况，qRT-PCR检测成骨因子Runx2、骨钙素（OCN）及Smad1、Smad5 mRNA表达，Western blot检测Smad1/5磷酸化水平及Smad1/5/8总蛋白表达。结果 OBI＋DEXA组ALP活性明显较OBI组低（P＜0.001），OBI＋DEXA＋ASD组ALP活性明显较OBI＋DEXA升高（P＜0.01）。OBI＋DEXA干预组成骨相关因子Runx2、OCN、Smad1和Smad5 mRNA表达较OBI组均明显下调（P＜0.01），OBI＋DEXA＋ASD组Runx2、OCN、Smad1和Smad5 mRNA表达较OBI＋DEXA干预组明显上调（P＜0.01和P＜0.05）。三组Smad1/5/8总蛋白表达差异无统计学意义，OBI＋DEXA干预组pSmad1/5较OBI干预组表达降低，OBI＋DEXA＋ASD干预组pSmad1/5表达较OBI＋DEXA干预组增加。结论 ASD可促进GC环境下小鼠BMSCs成骨分化，其机制可能与激活BMP/Smad信号通路有关。 |
| 英文摘要: |
| Objective To investigate the effect of akebia saponin D (ASD) on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) under glucocorticoid (GC) condition in mice. Methods Mice BMSCs were cultured in vitro. BMSCs were divided into three groups, and were induced by osteoblastic differentiation induction (OBI), OBI + DEXA (0.1 μmol/L), and OBI + DEXA (0.1 μmol/L) + ASD (10μmol/L), respectively. ALP activity was examined with ALP kit. Alizarin red staining was performed to determine the status of mineralization. qRT-PCR was performed to examine the mRNA expression of osteogenic factors Runx2, osteocalcin (OCN), Smad1, and Smad5. The phosphorylation level of Smad1/5 and expression of total Smad1/5/8 protein were examined using Western blotting. Results Compared to that in OBI group, ALP activity decreased in OBI + DEXA group (P＜0.001), but increased in OBI + DEXA (0.1 μmol/L) + ASD group (P＜0.01). Relative expression of osteogenesis related factors Runx2, OCN, Smad1, and Smad5 mRNAs were down-regulated in OBI + DEXA group versus OBI, but were up-regulated in OBI + DEXA + ASD group compared to OBI + DEXA group (P＜0.01 and P＜0.05). There was no obvious difference in level of Smad1/5/8 total protein between any two groups. pSmad1/5 level was obviously lower in OBI＋DEXA (0.1 μmol/L) group than in OBI group. It was strikingly higher in OBI + DEXA + ASD group than in OBI + DEXA group. Conclusion ASD promotes osteogenic differentiation of mice BMSCs under GC condition probably through activating BMP/Smad signaling pathway. |
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