Indian Hedgehog在牵张力促进成骨细胞增殖中的作用研究

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[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。

[关键词] 成骨细胞； Hedgehog； 牵张； 增殖

[中图分类号] R 780.2 [文献标志码] A [doi] 10.3969/j.issn.1000-1182.2012.03.003

indian hedgehog signaling is involved in the stretch induced proliferation of osteoblast Han Lei1, Zhang Xiao-ling2, Tang Guohua3. （1. Dept. of Orthodontics, Institute and Hospital of Stomatology, Nanjing University Medical School, Nanjing 210025, China; 2. Orthopaedic Cellular and Molecular Biology Laboratory, Institute of Health Scien-ces, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 3. Dept. of Orthodontics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China）

[Abstract] Objective To observe the role of the Hedgehog（Hh） genes in the proliferation of osteoblasts upon me-

chanical 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 inhi-

bitor 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.

[Key words] osteoblast； Hedgehog； stretch； proliferation

牙槽骨和颅颌面骨是应力丰富区，也是成骨细胞活跃区。在外界机械应力刺激下，成骨细胞的增殖分化是骨组织发生适应性改建和骨再生重建的源泉[1]。越来越多的实验证实牵张力是有效促进成骨细

胞活性和功能的力学刺激。现有的研究发现：与力学有关的信号包括细胞外调节蛋白激酶-1/2（extracel-lular regulated protein kinases-1/2，ERK-1/2）信号通路、c-Jun氨基末端激酶（c-Jun N-terminal kinases，