[刺猬信号通路与成骨细胞的增殖和分化相关]
[Hedgehog pathway is associated with the proliferation and differentiation of osteoblasts].
作者信息
Han Lei, Zhang Xiao-ling, Li Xuan, Tang Guo-hua
机构信息
Department of Orthodontics, College of Stomatology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China.
出版信息
Shanghai Kou Qiang Yi Xue. 2009 Jun;18(3):287-90.
PURPOSE
The purpose of this study was to evaluate the effects of Hedgehog signaling pathway on osteoblast proliferation and differentiation.
METHODS
Primary osteoblasts harvested from newborn rat calvarial bone were cultured for 21 days. The temporary expression of Shh and Ihh was detected by RT-PCR. The culture medium was then treated with either a recombinant Shh N-terminals protein (N-Shh) or cyclopamine (cy), a Hedgehog inhibitor. The proliferation of osteoblasts was quantified by MTT and FCM. The osteoblast differentiation and matrix mineralization were evaluated by ALP and Alizarin-Red staining. The expression of Ptch and Smo was quantified by real-time PCR.SAS 8.0 software package was used for statistical analysis.
RESULTS
Osteoblast expressed both Shh and Ihh. The expression level of Shh decreased with culture time while the amount of Ihh expression increased. N-Shh treatment increased the cell population and the number of cells in S phase (P<0.05. N-Shh also promoted ALP activities (P<0.05) and bone matrix calcification. Moreover, N-Shh application led to an increase in Ptch and Smo mRNA level (P<0.05). Treatment with cy, on the other side, inhibited osteoblast proliferation and differentiation(P<0.05).
CONCLUSION
Heghehog signaling pathway is actively involved in regulating osteoblast proliferation and differentiation.
目的
本研究旨在评估刺猬信号通路对成骨细胞增殖和分化的影响。
方法
从新生大鼠颅骨获取原代成骨细胞并培养21天。通过逆转录聚合酶链反应（RT-PCR）检测音猬因子（Shh）和印度刺猬因子（Ihh）的瞬时表达。然后用重组Shh N端蛋白（N-Shh）或刺猬信号通路抑制剂环杷明（cy）处理培养基。通过噻唑蓝（MTT）和流式细胞术（FCM）对成骨细胞的增殖进行定量分析。通过碱性磷酸酶（ALP）和茜素红染色评估成骨细胞分化和基质矿化。通过实时聚合酶链反应对帕奇蛋白（Ptch）和 smoothened蛋白（Smo）的表达进行定量分析。使用SAS 8.0软件包进行统计分析。
结果
成骨细胞表达Shh和Ihh。Shh的表达水平随培养时间下降，而Ihh的表达量增加。N-Shh处理增加了细胞数量和S期细胞数量（P<0.05）。N-Shh还促进了碱性磷酸酶活性（P<0.05）和骨基质钙化。此外，应用N-Shh导致Ptch和Smo mRNA水平升高（P<0.05）。另一方面，用cy处理抑制了成骨细胞的增殖和分化（P<0.05）。
结论
刺猬信号通路积极参与调节成骨细胞的增殖和分化。
Zotero 插件