摘要
目的观察不同浓度的氟对体外培养大鼠成骨细胞内钙调蛋白(CaM)表达的影响。方法采用骨组织块法分离培养新生Wistar大鼠颅骨成骨细胞,然后将不同浓度的氟作用于大鼠成骨细胞,染氟培养48 h后,采用半定量RT-PCR方法分析CaM基因的表达,免疫细胞化学法检测CaM蛋白的表达,采用生物化学的方法检测超氧化物歧化酶(SOD)、超氧阴离子自由基、丙二醛(MDA)的含量。结果与对照组比较,各氟组CaM基因及蛋白的表达均显著增加;与对照组相比,各氟组成骨细胞内SOD含量明显降低,超氧阴离子自由基含量明显增高,差异均具有统计学意义,而MDA含量无明显变化。结论氟在一定浓度范围内可引起培养成骨细胞的SOD含量明显降低、超氧阴离子自由基含量明显增高,且对成骨细胞的CaM蛋白的表达具有促进作用。
Objective To study the effects of different concentrations of fluoride on expression of calmodulin （ CaM ） in rat osteoblasts cultured in vitro. Methods Osteoblasts were isolated from the calvariae of new - born Wistar rats by means of bone tissue block, and cultured with DMEM. Different concentrations of fluoride were added to rat osteoblasts and incubated for 48 hours. The expression of CaM gene was studied by reverse transcription polymerase chain reaction （ RT - PCR ）. The expression of CaM protein was analyzed with immunohistochemical stain. The levels of superoxide dismutase （ SOD ） , malondialdehyde （MDA） , superoxide anion radical were tested by biochemical techniques. Results The expressions of CaM gene and protein were significantly higher in all excessive fluoride groups than that in control groups （ P 〈0.05）. The activity of SOD gradually enhanced with the elevation of fluoride, and was higher than that in control group （ P 〈 0.05 ）. The level of superoxide anion radical in cells of excessive fluoride groups was markedly increased than that in control groups （ P 〈 0.05 ）. There were no significant changes observed in the level of MDA of all different concentrations of fluoride groups compared with the control group. Conclusions The activity of SOD gradually enhanced and higher of superoxide anion radical were than that with the elevation of fluoride compared with the control group. The fluoride can stimulate the expression of CaM in osteoblasts in certain range of concentration.
出处
《中国地方病防治》
北大核心
2007年第5期333-336,共4页
Chinese Journal of Control of Endemic Diseases
基金
贵州省人才领导小组基金(编号Q2005-6)
贵州省国际合作项目(编号M2004-4)
关键词
氟
成骨细胞
钙调蛋白
Fluoride
Osteoblasts
Calmodulin
