摘要
目的观察Hedgehog(Hh)信号在牵张力调控成骨细胞增殖中的作用。方法体外分离和培养新生大鼠颅顶骨成骨细胞,应用Flexcell 4000TM加力系统施加3%和6%的2种牵张应变。用N端Hedgehog重组蛋白(N-Shh)、Hed-gehog通道抑制剂环巴胺(cy)和机械通道抑制剂三氯化钆(GdCl3)对成骨细胞进行干预。分别采用细胞计数、甲基噻唑基四唑(MTT)比色法和流式细胞仪检测成骨细胞的增殖。采用SAS 8.0软件包对结果进行统计分析。结果在成骨细胞体外培养过程中,牵张力促进了成骨细胞的增殖,以6%牵张应变更显著;牵张力仍能促进经cy处理后的成骨细胞的增殖,但这种促进作用能被GdCl3抑制。结论牵张力促进成骨细胞增殖的作用有一部分是通过上调Ihh的表达实现的。
Objective To observe the role of the Hedgehog（Hh） genes in the proliferation of osteoblasts upon mechanical tensile strains.Methods Primary osteoblasts harvested from newborn rat calvarial bone were subjected to 3% and 6% elongation of tensile stretches using Flexcell 4000TM strain unit.The cultures were also treated with either recombinant N-terminals Sonic Hedgehog（N-Shh） or cyclopamine（cy）,a Hh inhibitor or gadolinium（GdCl3）,an inhibitor of stretch-activated channels.The proliferation of osteoblasts was quantified by cell counting,methyl thiazolyl te-trazolium（MTT） and cell cycle detection via flow cytometry.Statistical analysis was performed using SAS 8.0 software package.Results The tensile strain,especially under 6% elongation,promoted osteoblast proliferation.Stretching force could also promote the proliferation even when the cells were treated with cy,but this effect was suppressed by GdCl3.Conclusion The induced proliferation of osteoblasts by mechanical stretched is mediated at least in part by Indian Hedgehog（Ihh） signaling.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2012年第3期234-238,共5页
West China Journal of Stomatology
基金
上海市科学技术委员会基金资助项目(08DZ2271100)
上海市重点学科建设基金资助项目(S30206)
