标题：皮质酮浓度对SD大鼠成骨细胞成骨基因表达的影响

      作者：康银辉 1，魏 波 1，祝兆波 1，郭伟雄 1，王朝军 1，宋丽君 2，林 颢 1，李广盛 1，楚佳奇 1，曾 荣 1
    (广东医学院附属医院骨科中心 1、生殖中心 2，广东 湛江 524001)

      卷次： 2015年26卷18期

      【摘要】 目的 探讨皮质酮对离体SD大鼠成骨细胞功能的影响。方法 采用多次胶原酶组织消化法获得
新生SD大鼠颅骨中的成骨细胞作为研究对象，用倒置显微镜及碱性磷酸酶、钙结节染色观察成骨细胞形态，
CCK-8法检测皮质酮对成骨细胞增殖的影响，根据CCK-8结果将SD大鼠成骨细胞分成四组，分别用含不同浓
度的皮质酮(0 ?mol/L、0.1 ?mol/L、1.0 ?mol/L、10.0 ?mol/L)的DMEM(H)培养基培养，作用24 h后，测定成骨细胞
内碱性磷酸酶含量，采用RT-PCR检测成骨细胞内COL1A、OCN、ALP基因表达，采用Western blot检测成骨细胞
内COL1A、OCN、RUNX-2蛋白的表达。结果 CCK-8结果显示皮质酮能抑制成骨细胞的增殖，皮质酮能抑制
成骨细胞内碱性磷酸酶的生成，与浓度明显相关；RT-PCR与Western blot检测可见COL1A、OCN、ALP基因表达
及COL1A、OCN、RUNX-2蛋白的表达下降。结论 皮质酮在超生理剂量的浓度下能抑制成骨细胞增殖及细胞
内碱性磷酸酶的合成，减少COL1A、OCN、ALP基因及COL1A、OCN、RUNX-2蛋白的表达。

      【关键词】 皮质酮；成骨细胞；基因表达；成骨

      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2015）18—2661—04


Effect of corticosterone concentration on osteoblastic gene expression of SD rat osteoblasts.
KANG Yin-hui 1, WEI
Bo 1, ZHU Zhao-bo 1, GUO Wei-xiong 1, WANG Chao-jun 1, SONG Li-jun 2, LIN Hao 1, LI Guang-sheng 1, CHU Jia-qi 1,
ZENG Rong 1. Orthopedic Centre 1, Lab of Reproductive Medicine 2, Affiliated Hospital of Guangdong Medical College,
Zhanjiang 524001, Guangdong, CHINA

【Abstract】 Objective To investigate the effect of different corticosterone concentration on the gene expres-
sion of SD rat osteoblasts in vitro. Methods Newborn SD rat osteoblasts in the skull were obtained by many times of
collagenase digestion as the object of study. The osteoblastic morphology was observed using inverted microscope, al-
kaline phosphatase and calcium nodes dyeing. And the effect of corticosterone concentration on osteoblastic prolifera-
tion was detected by CCK-8. According to the scores of CCK-8, the SD rat osteoblasts were divided into four groups
with different corticosterone concentrations of DMEM (H) culture (0 μmol/L, 0.1 μmol/L, 1.0 μmol/L, 10.0 μmol/L).
After 24 hours of DMEM (H) culture, content of osteoblastic alkaline phosphatase were measured, gene expressions
of osteoblastic COL1A, OCN, ALP were detected by RT-PCR, and the protein expression of osteoblastic COL1A,
OCN, RUNX-2 were measured by western blot. Results The scores of CCK-8 showed that corticosterone can re-
strain the proliferation of SD rat osteoblasts and the formation of osteoblastic alkaline phosphatase, which were signifi-
cantly related to concentration. And the results of RT-PCR and western blot both showed decreased expression of
COL1A, OCN, ALP gene and COL1A, OCN, RUNX-2 protein. Conclusion Corticosterone beyond physical dosage
can inhibit the proliferation of osteoblasts and synthesis of intracellular alkaline phosphatase, and reduce the expres-
sion of COL1A, OCN, ALP gene and COL1A, OCN, RUNX-2 protein.