摘要
目的观察氟对体外培养人成骨肉瘤细胞内骨粘连蛋白/骨连接素(osteonectin,ON)基因表达及氧化应激状态的影响。方法采用McCOY’s 5A培养基培养人成骨肉瘤细胞Saos-2。按照染氟剂量将骨肉瘤细胞Saos-2分为0(对照组)、0.625、1.250、2.500、5.000、10.000、20.000、40.000、80.000mg/L组。培养24h后收集细胞,实时荧光定量PCR(real-time RT-PCR)测定成骨细胞成骨相关基因ON mRNA的表达,用双标准曲线法计算基因表达的相对比值。氧化应激状态用化学比色法分别测定超氧化物歧化酶(superoxide dismutase,SOD)及丙二醛(malondialdehyde,MDA)的活性。结果 0.625、1.250、2.500、5.000mg/L组成骨肉瘤细胞ON mRNA表达分别为(0.014 3±0.032)、(0.057 3±0.02)、(0.042 8±0.017 7)、(0.076 2±0.005),低于对照组,差异有统计学意义(P均<0.01),但10.000、20.000mg/L组成骨肉瘤细胞ON mRNA表达分别为(11.6±1.01)、(12.7±1.18)、(1.65±0.00)、(1.97±0.46),明显高于对照组,差异有统计学意义(P<0.01),组间比较差异有统计学意义(F=305.842,P<0.01)。5.000～80.000 mg/L组SOD的活性分别为(94.14±3.16)、(63.36±3.63)、(53.00±4.26)、(55.50±5.89)、(53.46±6.40)×103 U/g,与对照组[(122.34±6.99)×103 U/g]比较明显降低,组间比较差异有统计学意义(F=21.734,P<0.01)。10.00mg/L组MDA的活性为(7.74±1.43)μmol/g,与对照组[(4.99±0.65)μmol/g]比较明显升高,组间比较差异有统计学意义(F=3.067,P<0.05)。成骨细胞内ON表达水平与SOD活力呈明显负相关。结论氟可能通过影响成骨细胞内骨连接素基因的表达和氧化应激状态而改变正常的成骨分化。
± Objective To observe osteonectin expression and the changes of oxidative stress in osteoblast treated with fluoride. Methods Human osteoblast cell （Saos-2） was cultured in McCOY＇s 5A medium and treated with fluoride （sodium fluoride, NaF）. There were eight groups including 0 （control group）, 0. 625, 1. 250, 2. 500, 5. 000, 10. 000, 20. 000, 40. 000,80. 000 mg/L groups. Expressions of osteonectin （ON） mRNA were detected by real-time PCR. Dual-standard curve method was used for analysis. Two oxidative stress indexes, superoxide dismutase （SOD） and malondialdehyde （MDA） were determined by measuring the absorbance using a micro titer plate reader. Results Expression of ON mRNA were lower in 0. 625, 1. 250, 2. 500, 5. 000 mg/L groups than the control group which were 0. 014 3±0. 032,0. 057 3 ±0.02, 0. 042 8±0. 017 7, 0. 076 2±0. 005, respectively （all P〈0. 01）; it was higher in the 10. 000,20. 000 mg/L groups than the control group at groups were significant （F = 305. 842, P 〈0.0 24 hours after 1）. Compared treated with fluoride. The differences among with the control group （122.34±6.99）, the SOD activity was decreased in groups treated with fluoride （5. 000-80. 000 mg/L groups, were respectively 94.14±3.16, 63.36±3.63, 53.00±4.26, 55.50±5.89, 53.46±6.40）, （all P 〈0.05）, the differences among groups were significant （F = 21. 734,P 〈0.01）. Compared with the control group （4. 99±0.65）, MDA activity of 10. 000 mg/L group （7.74±1.43） cells exposed to fluoride was significantly increased, the difference between groups was statistically significant （F = 3. 067, P 〈0.05）. ON expression in osteogenic cells was a clear negative correlation with SOD. Conclusion Fluoride may affect osteonectin expres sion and oxidative stress status of osteoblast and change the normal osteogenic differentiation.
出处
《新疆医科大学学报》
CAS
2012年第12期1627-1631,1637,共6页
Journal of Xinjiang Medical University
基金
国家自然科学基金(81102084)
关键词
氟中毒
基因表达
骨连接素
骨代谢
fluorosis
gene expression
osteonectin
bBone turn over
