于明香,王洪复,金慰芳,顾淑珠.137Cs γ射线对体外培养成骨细胞形态与几种细胞因子基因表达的影响[J].中华放射医学与防护杂志,2004,24(1):26-29
137Cs γ射线对体外培养成骨细胞形态与几种细胞因子基因表达的影响
Influence of 137Cs γ-irradiation on morphology and several cytokine mRNA expression of osteoblasts cultured in vitro
投稿时间:2003-03-28  
DOI:
中文关键词:  成骨细胞  电离辐射  基因表达
英文关键词:Osteoblasts  Radiation  Gene expression
基金项目:国家教委博士点科研基金资助项目(9936)
作者单位
于明香 200032, 复旦大学放射医学研究所骨代谢研究室 
王洪复 200032, 复旦大学放射医学研究所骨代谢研究室 
金慰芳 200032, 复旦大学放射医学研究所骨代谢研究室 
顾淑珠 200032, 复旦大学放射医学研究所骨代谢研究室 
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中文摘要:
      目的 观察137Csγ射线照射对体外培养成骨细胞的形态、核浆比与几种细胞因子基因表达的影响。方法 取第1代新生SD大鼠头盖骨成骨细胞传代后24h一次接受137Csγ射线照射,吸收剂量分别为10,25,50cGy和1,3,6,9,12Gy。观察各组成骨细胞形态与超微结构改变;Giemsa染色后进行显微形态计量,计算核浆比;细胞培养至14d抽提细胞内总RNA,RT-PCR方法半定量检测各组细胞IGF1,OPG,ODF基因mRNA表达量。结果 3Gy以上γ射线照射的细胞,胞体变大、变薄,细胞数减少,细胞内细胞器减少,溶酶体增多,核浆比降低,其改变随吸收剂量增加而突出。基因IGF1、OPG、ODF与OPG/ODFmRNA表达量在1Gy以上组随吸收剂量增加逐渐降低,与吸收剂量呈明显相关(P<0.05或0.01)。结论 137Csγ射线照射引起成骨细胞形态明显改变,增殖能力降低,核浆比降低,基因IGF-1、OPG、ODF与OPG/ODFmRNA表达量降低。
英文摘要:
      Objective To observe the influence of 137Csγ-irradiation on morphology and gene expression of osteoblasts cutured in vitro . Methods Osteoblasts were isolated from new-born SD rat crania and cultured in EMEM containing 10% NCS.The osteoblasts of the first passage were irradiated once with 137Csγ-rays after cultured for 24 hours.The irradiation doses were 10,25,50 cGy and 1,3,6,9,12 Gy,respectively.Their morphology and ultrastructure were observed under phase-contrast microscope and electronmicroscope.Karyoplasm ratio was determined after Giemsa staining.Total RNA was extracted with TRIzol reagent when the osteoblasts of the first passage were cultured for 14 days.mRNA expressions of insulin-like growth factor-1(IGF-1),osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) were determined with RT-PCR semi-quantitatively and compared among different groups. Results The osteoblasts irradiated with 137Csγ-rays over 3 Gy were larger,thinner,containing more nuclei,proliferating slowly, having fewer cell organelles and more lysosomes in the plasma,with decreased ratio of nucleus to plasma.All of the above changes were more obvious with increasing dosage of irradiation.Quantity of IGF-1,OPG,ODF and OPG/ODF mRNA expressions were decreased with increasing dosage of irradiation above 1 Gy.It was obviously correlated with the quantity of irradiation ( P<0.05 or 0.01,respectively). Conclusion By 137Csγ-irradiation osteoblasts can be induced to have morphological changes,decreased ability of proliferation,decreased karyoplasms ratio and decreased quantity of IGF-1,OPG,ODF and OPG/ODF mRNA expressions.
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