唑来膦酸对牙龈成纤维细胞增殖及凋亡的影响

Effect of zoledronic acid on cell proliferation and apoptosis of human periodontal fibroblasts

目的 观察低浓度的唑来膦酸对人牙龈成纤维细胞（human gingival fibroblasts,HGF）增殖及凋亡的影响,探讨唑来膦酸导致二膦酸盐相关性颌骨骨坏死（bisphosphonate-related osteonecrosisof the jaw,BRONJ）的发生机制.方法 利用不同浓度（0、0.5、1.0、5.0、10.0 μmol/L）唑来膦酸作用于HGF,24 h后流式细胞仪检测细胞凋亡率、甲基噻唑基四唑（methyl thiazolyl terazolium,MTT）法观察HGF增殖情况,利用细胞增殖度法动态检测第2、4、7天不同浓度唑来膦酸对HGF增殖的影响.结果 流式细胞仪检测结果显示唑来膦酸浓度为0、0.5、1.0、5.0、10.0μmol/L时细胞凋亡率分别为：（0.67±0.50）％、（2.13±0.21）％、（3.27±0.23）％、（4.17±0.35）％、（9.87±1.79）％.从1.0.0 μmol/L开始,随着唑来膦酸浓度的升高,细胞凋亡率显著升高（P＜0.05）.细胞增殖度法观察在0.5～ 10.0 μmol/L和7d范围内表现为浓度-时间依赖效应,随着浓度升高,时间延长,细胞增殖抑制更显著,不同浓度组之间吸光度值（A值）差异有统计学意义（P＜0.05）.结论 唑来膦酸对HGF具有抑制增殖和促进凋亡的作用.

Objective To observe the effect of different concentrations of zoledronic acid on human gingival fibroblast（HGF） cells proliferation and apoptosis, and to investigate the mechanism of bisphosphonate-related osteonecrosis of the jaw（BRONJ） caused by zoledronic acid.Methods Different concentrations（0, 0.5, 1.0, 5.0, 10.0 μmol/L） of zoledronic acid acted on the HGF.After 24 hours, flow cytometry was used to detect the rate of apoptosis and methyl thiazolyl terazolium（MTF） assay used to observe the proliferation of HGF.The effect of different concentrations of zoledronic acid on cell proliferation was examined by proliferation test on day 2, 4 and 7.Results Flow cytometry showed that when the concentration of zoledronic acid was 0, 0.5, 1.0, 5.0, 10.0 μmol/L, the apoptosis rate was （0.67± 0.50）%, （2.13±0.21）%, （3.27±0.23）%, （4.17±0.35）%, （9.87± 1.79）% respectively.The difference in HGF cells between control group and zoledronic acid groups was significant（P〈0.05） at three concentrations of zoledronic acid（1.0, 5.0, 10.0 μmol/L）.The cell proliferation test showed an concentration-time dependent effect.With increase of concentration and time, the cell proliferation was significantly inhibited.There was significant difference in absorbance value among the different concentration groups（P〈0.05）.Conclusions Zoledronic acid can inhibit HGF proliferation and promote its apoptosis.