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目的探讨丝裂原活化蛋白激酶(MAPKs)信号通路在调控苯并(a)芘[B(a)P]诱导人胚肺成纤维细胞(HELF)c-Jun活化中的作用。方法2.0μmol/L B(8)P处理HELF 0、3、6、12、24 h后或加入不同浓度B(a)P(0.0、0.5、1.0、2.0μmol/L)处理12 h后,通过免疫印迹法检测B(a)P对细胞c-Jun活性的影响;利用p38、c-Jun氨基末端激酶(JNK)以及细胞外调节蛋白激酶(ERK)的显性失活突变体分别阻断p38、JNK和ERK活性后,观察3种MAPKs信号分子与B(a)P诱导c-Jun活化之间的关系。结果在所观察的B(a)P作用时间和浓度范围内,c-Jun蛋白表达量无明显变化;B(a)P可诱导细胞内磷酸化c-Jun (Ser63/Ser73)水平增高,并随着作用时间的延长,细胞内c-Jun的磷酸化水平也逐渐增强,至12 h达到峰值(1.61±0.12,1.82±0.18),c-Jun磷酸化Set63、Ser73与actin灰度比值分别是对照组的20.1倍、15.2倍,作用24 h时细胞内c-Jun磷酸化水平呈现下降趋势,而且随着B(a)P浓度的增加,细胞内c-Jun的磷酸化水平也逐渐升高,呈现剂量-反应关系;使用显性失活突变体分别阻断JNK和ERK活性均可明显抑制B(a)P诱导细胞c-Jun磷酸化水平增加,但是阻断p38活性对B(a)P诱导细胞c-Jun磷酸化水平升高无明显影响。结论JNK和ERK信号通路调控B(a)P诱导的HELF细胞c-Jun活化,B(a)P促进c-Jun磷酸化的过程与p38信号通路无关。
Objective To investigate the role of mitogen-activated protein kinase (MAPKs) signaling pathway in the regulation of c-Jun activation induced by benzo (a) pyrene [B (a) P] in human embryonic lung fibroblasts. Methods After treated with 2.0μmol / L (8) P HELF for 0, 3, 6, 12 and 24 hours or with different concentrations of B (a) P (0.0,0.5,1.0 and 2.0μmol / L) The effect of B (a) P on the c-Jun activity was detected by Western blotting. The p38, p38 and c-Jun N-terminal kinase (JNK) and the dominant negative mutant of extracellular regulated protein kinase (ERK) JNK and ERK activity, observe the relationship between the three kinds of MAPKs signal molecules and B (a) P induced c-Jun activation. Results There was no significant change of c-Jun protein expression in the time and concentration range of B (a) P observed; B (a) P induced an increase in phosphorylated c-Jun (Ser63 / Ser73) The phosphorylation level of c-Jun increased gradually with the prolongation of action time, reaching the peak at 12 h (1.61 ± 0.12, 1.82 ± 0.18). The phosphorylation rates of c-Jun phosphorylated Set63, Ser73 and actin were The levels of c-Jun phosphorylation in the control group decreased by 20.1-fold and 15.2-fold, respectively, and the level of c-Jun phosphorylation increased gradually with the increase of B (a) P concentration (P <0.05), and the dose-response relation was shown. Using the dominant-negative mutant to block JNK and ERK activity separately inhibited B (a) P induced c-Jun phosphorylation, P induced cell c-Jun phosphorylation level had no significant effect. Conclusions The JNK and ERK signaling pathways regulate the c-Jun activation of HELF cells induced by B (a) P. The phosphorylation of c-Jun by B (a) P is not related to the p38 signaling pathway.