摘要
目的探讨抑制ERK1传导路径对支气管上皮细胞谷氨酰半胱氨酸合成酶(γ-GCS)合成的影响。方法用4-羟基壬烯醛(4-HNE)做刺激原,分为10μmol/L4-HNE、50μmol/LPD98059(MEK1抑制剂)孵育+10μmol/L4-HNE、正常对照组分别作用支气管上皮细胞0.5、2、4、8、12h后,检测细胞γ-GCSmRNA和蛋白的表达水平;10μmol/L4-HNE刺激细胞0.5、2、4、8、12h后,检测磷酸化JNK、P38MAPK、ERK1/2、活化蛋白-1(AP-1)活性;观察MEK1抑制剂PD98039对AP-1结合活性的影响。结果 10μmol/L4-HNE、50μmol/LPD98059孵育+10μmol/L4-HNE、正常对照组2、4、8、12h,细胞γ-GCS和γ-GCSmRNA表达三组差异有统计学意义,50μmol/LPD98059孵育+10μmol/L4-HNE较其他两组增加。10μmol/L4-HNE刺激0.5、2、4、8、12h组细胞磷酸化ERK1/2水平,AP-1结合活性较对照组增强。PD98059孵育后,4-HNE作用0.5、2、4、8、12h后,AP-1结合活性较未用PD98059孵育4-HNE刺激组降低。结论 MEK1抑制剂PD98059可以抑制支气管上皮细胞16-HBE细胞内ERK1-AP1信号传导通路,对4-HNE引起支气管上皮细胞γ-GCS合成有促进作用,阻断AP-1途径后支气管上皮细胞并不能减少γ-GCS的表达。
Objective To explore the effects of blocking AP-1 transduction pathway on the expression of γ-glutamylcysteine synthetase (γ-GCS) in the bronchial epithelium cell (16-HBE).Methods Bronchial epithelium cell (16-HBE) were stimulated by 10 μmol/L 4-HNE,50 μmol/L PD98059+10 μmol/L 4-HNE,control for 0.5,2,4,8,12 hours,and the γ-GCS and γ-GCS mRNA were measured.The phosphorylation of extracellular regulated proteinkinases(ERK)1/2,c-JUN N-terminal kinase (JNK),P38 (mitogen-activated protein kinases) MAPK and the combining activity of AP-1 after 10 μmol/L 4-HNE stimulating for 0.5,2,4,8,12 hours were all estimated,combining activity of AP-1 in 10 μmol/L 4-HNE,50 μmol/L PD98059 +10 μmol/L 4-HNE,control groups were measured by EMSA.Results The level of γ-GCS and γ-GCS mRNA in 10 μmol/L 4-HNE,50 μmol/L PD98059 +10 μmol/L 4-HNE,control groups after 4-HNE stimulating for 0.5,2,4,8,12 h were difference significantly,and the results of 50 μmol/L PD98059 +10 μmol/L 4-HNE were much higher than two others groups results.The level of phosphorylation of ERK,AP-1 combining activity in the 10 μmol/L 4-HNE stimulation for 0.5,2,4,8,12 hours groups were higher than the control groups.AP-1 combining activity in groups of pre-incubated with or without PD98059 before 4-HNE stimulation also had difference significantly.Combining activity of AP-1 in 50 μmol/L PD98059 +10 μmol/L 4-HNE were decreased comparing with 10 μmol/L 4-HNE groups and control groups.Conclusions PD98059 could increase the expression of γ-GCS treated 4-HNE,though it could decrease the AP-1 combining activity.Blocking AP-1 transduction pathway could not inhabit the synthesis of γ-GCS in the bronchial epithelium cell.
出处
《中华临床医师杂志（电子版）》
CAS
2011年第7期1967-1972,共6页
Chinese Journal of Clinicians(Electronic Edition)
