尼古丁对PDLFs细胞MAPKs信号通路影响

Nicotine-induced periodontal ligament fibroblasts (PDLFs) apoptosis through MAPKs signal pathway

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 study the effect and mechanism of mitogen-activated protein kinase（MAPK） signal transduction pathway in nicotine-induced human periodontal ligament fibroblasts（PDLFs） apoptosis. Methods The PDLFs were treated with different doses of nicotine（1,10,and 100 μg/mL）.Twenty-four hours after the treatments,the expressions of JAK1,JAK2,MAPK,MAPKK,JNK,p38,and ERK were detected with reverse transcriptase polymersase chain reaction（RT-PCR） and the expression of caspase-3 was determined with western blot. Results After the treatments of 1,10 and 100 μg/mL nicotine,the transcriptive expressions of the genes were 65.17±8.45,106.17±22.22,and 115.50±8.12 for JAK1,103.00±13.13,118.17±13.17,and 159.00±13.74 for JAK2,126.69±18.20,143.02±13.97,and 172.64±26.43 for MAPKK,79.17±4.49,115.67±7.66,and 122.33±8.41 for MAPKK,110.64±11.14,128.54±11.72,and 132.90±11.99 for p38,106.16±26.89,123.17±13.09,and 132.68±11.97 for JNK,131.52±19.85,144.76±13.87,and 153.59±13.85 for caspase-3,and 128.33±13.50,144.00±12.53,and 153.00±12.77 for caspase-3 translation,respectively.Compared with those of the control group,all the differences were statistically significant（P0.05 or P0.01 for all）.The expressions of the genes showed a gradual increasing trend with the increment of nicotine dose,except the transcription of ERK. Conclusion Nicotine-induced PDLFs apoptosis may not be through the ERK-MAPK pathway but through p38-MAPK and JNK-MAPK pathway and nicotine could lead to the increase of caspase-3.