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| 强骨饮调节破骨细胞外泌体影响成骨细胞分化的初步研究 |
| Preliminary study of the effect of the strong-bone decoction on osteoblast differentiation by regulation of osteoclast-derived exosomes |
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| DOI：10.3969/j.issn.1006-7108.2020.11.007 |
| 中文关键词: 中医中药 破骨细胞 成骨细胞 外泌体 β环状蛋白 |
| 英文关键词:Chinese traditional medicine osteoclast osteoblast exosomes β-catenin |
| 基金项目:浙江省中医药“十三五”重点学科项目（2017-XK-A16） |
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| 中文摘要: |
| 目的 初步探究强骨饮调节破骨细胞来源外泌体影响成骨细胞分化的内在机制。方法 制备强骨饮含药血清，构建破骨分化和成骨分化模型。设置对照血清组和含药血清组，共同干预破骨细胞分化，找到最佳含药血清浓度。采用免疫印迹法检测两组细胞外泌体释放情况。将两组外泌体加入成骨分化模型中，采用化学发光法检测碱性磷酸酶（ALP）含量，采用实时荧光定量测量成骨特异性转录因子（Runx2）、骨桥蛋白（OPN）、骨钙素（OCN）表达情况。最后运用免疫印迹法检测强骨饮干预下破骨分化的β环状蛋白表达以及外泌体作用成骨分化的β环状蛋白表达。结果 细胞染色显示含药血清可明显抑制TRAP活性。在两组破骨分化模型外泌体中均表达CD9、CD63、CD81蛋白。在两组外泌体干预下，ALP、Runx2、OPN和OCN的表达较对照组均降低，但含药血清组中成骨因子表达较对照血清组增加。两组成骨分化模型在外泌体干预下β环状蛋白检测较对照组均减少，但含药血清组β环状蛋白较对照血清组增加。含药血清组破骨细胞检测β环状蛋白较对照血清组增加。结论 强骨饮可抑制破骨分化，改善破骨来源外泌体对成骨分化的抑制作用。破骨来源外泌体可能传递miRNA作用wnt/β-catenin通路，影响成骨细胞分化。 |
| 英文摘要: |
| Objective To preliminarily explore the mechanism of the effect of the strong-bone decoction on osteoblast (OB) differentiation by regulating osteoclast (OC)-derived exosomes. Methods The serum containing of the strong-bone decoction was prepared and the differentiation models of OC and OB were constructed. The control-serum group and drug-serum group were set up. The best drug containing serum concentration for intervening the OC differentiation was discovered. The exosomes release was detected using Western blotting (WB) method. ALP contents were detected with chemiluminescence method. The expressions of runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) were measured with real-time PCR. Finally, the expression of β-catenin in OC and OB was detected wit WB. Results The cell staining showed that the activity of TRAP was significantly inhibited in the drug-serum group. CD9, CD63, and CD81 were expressed in exosomes of OC differentiation model in both groups. The expressions of ALP, Runx2, OPN and OCN in both intervening groups were lower than those in the control group. However, these osteogenic factors expressed more in the drug-serum group than in the control-serum group. The expression of β-catenin in OB in both groups was lower than that in the control group. However, β-catenin in OB expressed more in the drug-serum group than in the control-serum group. β-catenin in OC expressed more in the drug-serum group than in the control-serum group. Conclusion The strong-bone decoction inhibits the differentiation of OC and improves the negative effect of osteoclast-derived exosomes on OB differentiation. The osteoclast-derived exosomes may transfer miRNA to act on wnt/β-catenin pathway and influence OB differentiation. |
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