【摘要】 目的本研究旨在探讨细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号通路在组胺诱导的大鼠气道平滑肌细胞(airway smooth muscle cells,ASMCs)增殖中的调控作用。方法以体外分离、培养的ASMCs为研究对象,加入不同浓度组胺(histanmine,Hist)和PD98059对其进行处理,采用四甲基偶氮唑蓝(MTT)微量比色法检测Hist及ERK信号通路对ASMCs增殖的影响,Western blot检测ERK1/2、磷酸化ERK1/2(phosphorylated-ERK1/2,p-ERK1/2)蛋白的表达,采用酶联免疫吸附试验(ELISA)测定ASMCs核内NF-κB水平,并对各组进行比较。结果低浓度和高浓度的Hist均能不同程度刺激ASMCs增殖,100μmol/L时细胞存活率达201.3%;Hist组ASMCs增殖反应与对照组相比显著增加,而Hist+PD980059组ASMCs的增殖反应显著低于Hist组;Hist组ASMCs中的p-ERK1/2蛋白表达较正常对照组明显增强,在加入PD980059后Hist诱导的活化ERK1/2相对于Hist组表达显著下降;Hist处理组NF-κB吸光度值显著高于对照组,Hist+PD98059组NF-κB吸光度值较Hist处理组显著降低。结论Hist可显著诱导ASMCs增殖,ERK信号通道在此调控中起重要作用,而且还可能参与ASMCs中Hist诱导的NF-κB活化。更多还原

【Abstract】 Objective This work was designed to explore the regulation of extracellular signal-regulated kinase(ERK)signaling pathway in histamine-induced rat airway smooth muscle cell proliferation.Methods ASMCs were isolated and cultured in vitro and then exposed to different doses of histanmine(Hist)and PD98059.The proliferative effect of ASMCs was measured using MTT microcolorimetric assay.The expression of ERK1/2 and phosphorylated ERK1/2 proteins were detected by western blot.ELISA was used to evaluate the levels of NF-κB in ASMCs,and then compared with each group.Result Low and high concentration of Hist were able to stimulate varying degrees of ASMCs proliferation,100μmol /L when the cell survival rate reached 201.3%.Compared with control group,ASMCs proliferation in Hist group was increased significantly,while Hist + PD980059 group was lower than the latter one.The expression of p-ERK1/2 protein in Hist group increased markedly than the control group.Joined with PD980059,the expression of actived ERK1/2 in Hist group dropped significantly.The absorbance value in Hist group was higher than the control group,while Hist + PD98059 group was lower than the former one.Conclusion Histanmine can induce proliferation of ASMCs significantly,not only did ERK signal channel play an important role in regulating histamine-induced rat ASMCs proliferation,but also may be involved in the Hist-induced NF-κB activation.更多还原