尼古丁作用下牙周膜细胞c-Fos通路表达变化的相关研究

Effect of nicotine on c-Fos signaling pathway of periodontal ligament cells

目的探索尼古丁作用于人牙周膜成纤维细胞(human periodontal ligam ent fibroblasts,hPDLF)后c-Fos通路相关因子的表达变化,为揭示其参与吸烟相关性牙周炎的可能功能作用提供理论依据。方法改良组织块法培养hPDLF,时间和浓度梯度实验选择理想尼古丁作用时间及浓度,qPCR及蛋白印迹实验分别观察尼古丁和/或α-银环蛇毒素(α-Bungatoxin,α-BTX)作用于牙周膜细胞后c-Fos mRNA及蛋白表达。结果尼古丁作用于hPDLF后,c-FosmRNA和蛋白表达上调,变化具有浓度和时间依赖性。10-5 M尼古丁作用于牙周膜细胞1 h后,c-Fos mRNA及蛋白的表达上调最为显著,差异具有统计学意义(P<0.05)。尼古丁特异型受体烟碱型乙酰胆碱受体α7亚型(α7 acetylcholine receptor,α7 nAChR)特异性拮抗剂α-BTX可显著逆转这一作用,差异具有统计学意义(P<0.05)。结论尼古丁可能通过α7 nAChR调节c-Fos表达及活性从而在吸烟相关性牙周炎中发挥破坏作用。

Objective To explore the changes of expressions of c-Fos after nicotine treatment of human periodontal ligament fibroblasts(hPDLFs)in order to help find out about the mechanism of smoking related periodontitis.Methods hPDLFs were cultured using tissue block methods.The optimal length and concentration of treatment were studied via gradient experiments before qPCR and Western blot were performed to observe the changes of expressions of mRNA and protein in c-Fos after treatment with nicotine and/orα-bungatoxin(α-BTX).SPSS software was used to analyse the data.Results After nicotine treatment,the mRNA and protein expressions of c-Fos in hPDLFs changed in a time-and concentration-dependent manner.After one hour of treatment with 10-5 M nicotine,the expressions of c-Fos in hPDLFs were increased with statistically significant significance at both mRNA and protein levels(P<0.05).The specific antagonist of nicotine,theα7 acetylcholine receptor(α7 nAChR),could significantly reverse this effect(P<0.05).Conclusion Nicotine can regulate the expression and function of c-Fos in hPDLFs throughα7 nAChR;thus participating in the development of smoking related periodontitis.