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目的:研究脂肪细胞中氧自由基(ROS)对纤溶酶原激活物抑制物-1(PAI-1)表达的调节,并探讨其机制。方法:培养3T3-L1细胞,并诱导其分化成为脂肪细胞,以MTT比色法检测细胞的活性。分别以定量PCR、多重免疫分析及夹心ELISA法检测PAI-1 mRNA和蛋白表达的水平,并采用多重磷酸化蛋白分析系统检测细胞内多种信号分子的蛋白磷酸化水平。结果:H2O2可剂量依赖性地增加PAI-1的产生。并且激活了3T3-L1脂肪细胞中多种信号转导通路,包括ERK1/2、JNK、Akt、p70 S6K及JAK/STAT,其中Akt、JAK/STAT及ERK1/2的活化可能参与到H2O2对于PAI-1的调节过程中。结论:H2O2可能通过磷酸化激活Akt、JAK/STAT及ERK1/2,上调脂肪细胞PAI-1的表达。
AIM: To investigate the regulation of the expression of plasminogen activator inhibitor-1 (PAI-1) by oxygen free radicals (ROS) in adipocytes and to explore its mechanism. Methods: The 3T3-L1 cells were cultured and induced to differentiate into adipocytes. The cell viability was measured by MTT assay. The levels of PAI-1 mRNA and protein were detected by quantitative PCR, multiplex immunoassay and sandwich enzyme-linked immunosorbent assay (ELISA). The phosphorylation levels of PAI-1 mRNA and protein were detected by multiplex phosphoprotein assay system. Results: H2O2 increased PAI-1 production in a dose-dependent manner. Activation of Akt, JAK / STAT and ERK1 / 2 may be involved in the activation of H2O2 in PAI (P <0.05), and activation of various signal transduction pathways in 3T3-L1 adipocytes including ERK1 / 2, JNK, Akt, p70 S6K and JAK / -1 adjustment process. Conclusion: H2O2 may up-regulate the expression of PAI-1 in adipocytes through phosphorylation of Akt, JAK / STAT and ERK1 / 2.