槟榔碱对人牙周膜成纤维细胞凋亡的影响

目的 研究槟榔碱(Arecoline)对体外培养的人牙周膜成纤维细胞(human periodontal ligament fibro-blast,hPDLFs)中凋亡及相关蛋白p-JNK、p-p53、Bcl-2表达的影响.方法 采用组织块培养法培养原代hPDLFs,并传代纯化后用于实验.采用浓度为0μg/mL、20μg/mL、40μg/mL、80μg/mL的槟榔碱处理牙周膜成纤维细胞12 h,通过MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,蛋白免疫印记法检测槟榔碱对hPDLFs中p-JNK、p-p53、Bcl-2表达的情况.结果 槟榔碱作用于hPDLFs后,细胞增殖受到抑制,细胞凋亡率随着药物浓度增加而增加,p-JNK、p-p53蛋白表达逐渐增强,Bcl-2蛋白表达逐渐降低,呈现浓度依赖性.结论 提示槟榔碱可通过激活JNK和p53的磷酸化水平导致hPDLFs的凋亡,对牙周组织的再生有抑制作用.

Effects of arecoline on apoptosis of human periodontal ligament fibroblasts

Objective To observe the effects of arecoline on the related protein expressions in the apoptosis of cul-tured human periodontal ligament fibroblasts (hPDLFs), such as p-JNK, p-p53, and Bcl-2. Methods hPDLFs were iso-lated from human periodontal ligament tissues and expanded in vitro. hPDLFs were treated with different concentrations of arecoline (0, 20, 40 and 80 μg/mL) for 12 h. The proliferative activities were evaluated by MTT. The expressions of p-JNK, p-p53, and Bcl-2 were determined by Western blot. Results Arecoline inhibited cell proliferation and induced apoptosis protein. The protein level of Bcl-2 was decreased, while p-JNK and p-p53 were increased (P<0.05). The pro-tein expressions were in a concentration-dependent manner with arecoline. Conclusion It demonstrates arecoline might contribute to the apoptosis of hPDLFs, and could destroy periodontal tissues.