氧自由基增强脂肪细胞IL-6表达的实验研究

Reactive oxygen species increase the expression of interleukin-6 in adipocytes

目的:研究脂肪细胞中氧自由基(ROS)对白细胞介素-6(IL-6)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的mRNA和蛋白表达的水平。采用抑制剂干预的方法,寻找影响IL-6表达的信号分子。结果:作为一种重要的ROS,H2O2可剂量依赖性的增加IL-6的产生。Akt、JAK/STAT及ERK1/2的活化可能参与到H2O2对于IL-6的调节过程中。结论:ROS可能通过磷酸化激活Akt、JAK/STAT及ERK1/2,上调脂肪细胞IL-6的表达,从而参与肥胖、心血管疾病以及肿瘤等病理生理过程。

Objective：To investigate the regulatory effects of ROS （reactive oxygen species） on the production of IL-6 （interIukin-6）, and try to determine the signaling cascades involved in it. Methods：3T3-L1 ceils were cultured and differentiated into mature adipocytes. Cell viability was measured by MTT. The mRNA expression level of IL-6 was evaluated by quantitative real-time PCR. Quantification of the IL-6 protein levels secreted into conditioned medium was performed by multiplex immunoassay and sandwich ELISA. The protein kinases associated with the ROS-induced increase of IL-6 were sceened by intervention of specific inhibitors. Results： In 3T3-L1 adipocytes, H2O2 significantly augmented the expression of IL-6. Akt, JAK/STAT and ERK1/2 may participate in the H2O2-induced increase of IL-6. Conlusion： H2O2 markedly up-regulates the production of IL-6 in 3T3-L1 adipocytes via some intracellular signal pathways such as Akt, JAK/STAT and ERK1/2, which may be involved in the pathophysilology of obesity, caidiovascular diseases and cancer.