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目的:探讨不同浓度尼古丁对人牙周膜成纤维细胞(PDLFs)增殖及纤维结合蛋白(Fn)合成的作用。方法:不同浓度的尼古丁(50 ng/ml,250 ng/ml,500 ng/ml,1μg/ml,2μg/ml,3μg/ml)作用于PDLFs,MTT比色法检测细胞增殖活性,流式细胞仪检测细胞周期,酶联免疫吸附测定法(ELISA)检测Fn合成含量。结果:不同浓度尼古丁作用下:人PDLFs的增殖均被抑制,且呈浓度依赖性。浓度为250 ng/ml~3 ug/ml的尼古丁有明显抑制人PDLFs增殖的作用(P<0.05),3 ug/ml尼古丁显示出最强的抑制增殖作用(P<0.01);人PDLFs的G0-G1期、S期、G2-M期与对照组相比,G0-G1期细胞周期分布比例逐渐增高,S期和G2-M期逐渐降低,差异均有统计学意义,呈浓度依赖性;人PDLFs合成Fn逐渐减少,呈浓度依赖性。浓度为50 ng/ml~3μg/ml的尼古丁均有明显抑制人PDLFs合成Fn的作用(P<0.05),其中3μg/ml抑制作用最强。结论:尼古丁抑制牙周膜细胞的增殖及Fn的合成,呈浓度依赖性,并影响其细胞周期的进程,进而影响牙周新附着的形成,加重牙周病的病情。
AIM: To investigate the effects of different concentrations of nicotine on the proliferation and fibronectin (Fn) synthesis of human periodontal ligament fibroblasts (PDLFs). Methods: PDLFs were treated with different concentrations of nicotine (50 ng / ml, 250 ng / ml, 500 ng / ml, 1 μg / ml, 2μg / ml and 3μg / ml). MTT assay was used to detect the cell proliferation. Flow cytometry The cell cycle and enzyme-linked immunosorbent assay (ELISA) were used to detect the Fn content. Results: Under the different concentrations of nicotine, the proliferation of human PDLFs was inhibited in a concentration-dependent manner. Nicotine at a concentration of 250 ng / ml ~ 3 ug / ml significantly inhibited the proliferation of human PDLFs (P <0.05), nicotine at 3 μg / ml showed the strongest inhibitory effect on proliferation (P <0.01) -G1, S and G2-M phases compared with the control group, the proportion of cell cycle distribution in G0-G1 phase increased gradually, the S phase and G2-M phase decreased gradually, the differences were statistically significant and in a concentration-dependent manner; Human PDLFs synthesis Fn gradually decreased in a concentration-dependent manner. Nicotine at a concentration of 50 ng / ml ~ 3 μg / ml significantly inhibited the synthesis of Fn by human PDLFs (P <0.05), of which 3 μg / ml had the strongest inhibitory effect. CONCLUSION: Nicotine can inhibit the proliferation and Fn synthesis of periodontal ligament cells in a concentration-dependent manner and affect the process of cell cycle, which may affect the formation of new periodontal attachment and aggravate the condition of periodontal disease.