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目的探讨尼古丁诱导人牙周膜成纤维细胞(PDLFs)凋亡过程中丝裂素活化蛋白激酶(MAPK)信号转导通路的作用及作用机制。方法采用不同浓度的尼古丁(1、10和100μg/mL)作用于PDLFs细胞24 h后,使用逆转录聚合酶链反应(RT-PCR)测定JAK 1、JAK 2、MAPKK、MAPK、JNK、p 38和ERK的基因表达水平;Western blot检测caspase-3蛋白的表达水平。结果浓度分别为1、10和100μg/mL的尼古丁作用于PDLFs细胞24 h后,JAK 1转录水平表达量分别为(65.17±8.45)、(106.17±22.22)和(115.50±8.12),JAK 2转录水平表达量分别为(103.00±13.13)、(118.17±13.17)、(159.00±13.74),MAPK转录水平表达量分别为(126.69±18.20)、(143.02±13.97)、(172.64±26.43),MAPKK转录水平表达量分别为(79.17±4.49)、(115.67±7.66)、(122.33±8.41),p 38转录水平表达量分别为(110.64±11.14)、(128.54±11.72)、(132.90±11.99),JNK转录水平表达量分别为(106.16±26.89)、(123.17±13.09)、(132.68±11.97),Caspase 3转录水平表达量分别为(131.52±19.85)、(144.76±13.87)、(153.59±13.85),Caspase 3蛋白翻译水平表达量分别为(128.33±13.50)、(144.00±12.53)、(153.00±12.77),与正常对照组比较,差异均有统计学意义(P<0.05或P<0.01);随着尼古丁剂量的增加,这些基因表达均呈逐渐增高趋势;而ERK的表达水平则无明显变化。结论尼古丁通过介导p 38-MAPK和JNK-MAPK通路诱导人PDLFs细胞凋亡,并导致凋亡执行蛋白Caspase 3的表达增加。
Objective To investigate the role and mechanism of mitogen-activated protein kinase (MAPK) signal transduction pathway induced by nicotine in human periodontal ligament fibroblasts (PDLFs). Methods PDLFs cells were treated with different concentrations of nicotine (1, 10 and 100 μg / mL) for 24 h, and JAK1, JAK2, MAPKK, MAPK, JNK and p38 were determined by reverse transcriptase polymerase chain reaction And ERK gene expression; Western blot detection of caspase-3 protein expression levels. Results The expression levels of JAK 1 at 1, 10 and 100 μg / mL of nicotine for PDLFs cells for 24 h were (65.17 ± 8.45), (106.17 ± 22.22) and (115.50 ± 8.12), respectively. JAK 2 transcription The expression levels of MAPK were (126.69 ± 18.20), (143.02 ± 13.97) and (172.64 ± 26.43), respectively. The levels of MAPKK transcripts were (103.00 ± 13.13), (118.17 ± 13.17) and (159.00 ± 13.74) The expression levels of p 38 in the expression level of p38 were (110.17 ± 4.49), (115.67 ± 7.66) and (122.33 ± 8.41) The expression level of Caspase 3 was (106.16 ± 26.89), (123.17 ± 13.09) and (132.68 ± 11.97), respectively. The expression levels of Caspase 3 were (131.52 ± 19.85), (144.76 ± 13.87) and (153.59 ± 13.85) The levels of translational expression of Caspase 3 were (128.33 ± 13.50), (144.00 ± 12.53) and (153.00 ± 12.77), respectively, which were significantly different from those of the normal control group (P <0.05 or P <0.01) The nicotine dose increased, the expression of these genes showed a gradual increase trend; while the expression level of ERK had no significant change. Conclusion Nicotine induces the apoptosis of human PDLFs through the mediation of p38-MAPK and JNK-MAPK pathways, leading to the increase of the expression of Caspase 3.