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| FSHβ在BMP9诱导间充质干细胞成骨分化中的作用研究 |
| The study on the role of FSHβ in BMP9-induced osteogenic differentiation of mesenchymal stem cells |
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| DOI：10.3969/j.issn.1006-7108.2019.06.010 |
| 中文关键词: 骨形态蛋白9 卵泡刺激素 间充质干细胞 Notch DAPT |
| 英文关键词:bone morphogenetic protein 9 follicle stimulating hormone mesenchymal stem cells Notch DAPT |
| 基金项目: |
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| 中文摘要: |
| 目的 探讨卵泡刺激素β亚基(follicle-stimulating hormone β-subunit，FSHβ)
在骨形态发生蛋白9(bone morphogenetic protein 9，BMP9)诱导间充质干细胞(mesenchymal
stem cells，MSCs)成骨分化中的作用及可能的机制。方法 利用细胞化学染色方法检测处理
因素(重组腺病毒或γ促分泌酶抑制剂DAPT)作用于间充质干细胞5 d后细胞中碱性磷酸酶
(alkaline phosphatase，ALP)染色的变化。通过实时定量PCR(quantitative real-time
PCR，Q-PCR)检测BMP9 mRNA、FSHβ mRNA和Notch信号通路靶基因(Hey1) mRNA的表达水
平。利用茜素红S(alizarin red S，ARS)染色检测腺病毒感染间充质干细胞21 d后细胞中
钙盐的沉积情况。结果 FSHβ处理组ALP表达较GFP对照组无明显改变，BMP9处理组ALP
表达较GFP对照组明显增加，而BMP9与FSHβ联合处理组ALP表达较BMP9处理组显著增加。
另外，BMP9处理组晚期成骨标志物钙盐结节、Notch信号靶基因Hey1 mRNA表达水平较GFP
对照组显著增加(P＜0.01)，而BMP9与FSHβ联合处理组较BMP9处理组表达显著增加(P＜
0.01)。使用药物DAPT抑制Notch信号后，BMP9与DAPT联合处理组较BMP9单独处理组，ALP
染色、Hey1 mRNA表达显著降低(P＜0.05)；且BMP9+FSHβ+DAPT联合处理组较BMP9+FSHβ
联合处理组ALP染色、Hey1 mRNA表达也显著降低(P＜0.05)。结论 FSHβ可增强BMP9诱导
的间充质干细胞成骨分化，Notch信号通路可能在其中发挥重要作用。 |
| 英文摘要: |
| Objective To investigate the role of FSHβ (follicle-stimulating hormone β-subunit) in BMP9 (bone morphogenetic protein 9)-induced osteogenic differentiation of mesenchymal stem cells. Methods The cytochemical staining method was used to detect the change of ALP (alkaline phosphatase) expression in the cells on the 5th day after the treatment factors (recombinant adenovirus or γ-secretase inhibitor DAPT) were applied to MSCs. Quantitative real-time
PCR (Q-PCR) was used to detect the changes of mRNA expression levels of BMP9, FSHβ and Notch signaling pathway target genes (Hey1). Alizarin Red S staining was used to detect the deposition of calcium salts in the cells on the 21st day after adenovirus-infection of MSCs. Results The ALP staining of FSHβ treatment group was not significantly changed compared with the GFP group; the ALP staining of BMP9 treatment group was significantly higher than the GFP group; and the ALP activity of BMP9 and FSHβ treatment group was significantly higher than the BMP9 treatment group. In addition, the expression levels of late osteogenic markers (calcium salt nodules) and Notch signaling target gene Hey1 mRNA in the BMP9 treatment group were significantly higher than those in the GFP group (P<0.01), and further increased in the combined treatment group than the BMP9 treatment group (P<0.01). After inhibiting Notch signal by DAPT, the ALP staining and the expression of Hey1 mRNA in the combined treatment group of BMP9+DAPT were significantly reduced compared with the separate treatment group of BMP9 (P<0.05). Moreover, the ALP staining and the expression of Hey1 mRNA in BMP9+FSH +DAPT combined treatment group were significantly lower than that in BMP9+FSH combined treatment group (P<0.05). Conclusion FSHβ can enhance the osteogenic differentiation of mesenchymal stem cells induced by BMP9, and Notch signaling may play an important role in it. |
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