尼古丁诱导人牙周膜成纤维细胞凋亡的实验研究

Study on Apoptosis of Human Periodontal Ligament Fibroblasts Induced by Nicotine

目的:了解在体外培养环境中,尼古丁对人牙周膜成纤维细胞凋亡的影响。方法:将尼古丁作用于培养至第6代的人牙周膜成纤维细胞后,通过四唑盐比色法、流式细胞仪研究尼古丁能否诱导人牙周膜成纤维细胞凋亡。结果:四唑盐比色法证实当尼古丁浓度在0.05～4mg/L之间时,对牙周膜成纤维细胞的抑制呈浓度依赖性,流式细胞仪分析提示,当尼古丁浓度在0.5～4mg/L之间时,对牙周膜成纤维细胞的凋亡率呈浓度依赖性。结论:尼古丁对人牙周膜成纤维细胞有一定的毒性作用,通过抑制牙周膜成纤维细胞增殖和诱导凋亡加重对周膜的破坏。

Objective：To investigate the effects in the induction of apoptosis of nicotine on periodontal ligament fibroblasts（PDLF）in vitro.Methods：Human primary periodontal ligament fibroblasts were cultured in vitro,and the sixth passage cells were used in the experiments.PDLFs were cultured in the presence of nicotine at various concentrations.The growth of cell was measured by MTT method.The apoptosis and the cycle of the cell were examined by Flow Cytometer.Results：Nicotine can inhibited periodontal ligament fibroblasts proliferation significantly in a dose-dependent manner when the concentrations of nicotine were 0.05μg /ml to 4μg/ml.Apoptosis rate assay with flow cytometry showed that nicotine can promoted the apoptosis of periodontal ligament fibroblasts in a dose-dependent manner when the concentrations of nicotine were 0.5μg /ml to 4μg/ml.Conclusion：Nicotine has toxic effect to the ligament fibroblasts by inhibiting cell proliferation and inducing cell apoptosis,which may aggravate destruction of periodontal membrane in periodontal disease.