目的:检测脂多糖(LPS)和金黄色葡萄球菌(SAC)对人外周血单个核细胞(PBMC)IL-23和IL-12各亚基表达的调节作用并探索其作用机制。方法:用LPS和SAC直接刺激人PBMC,或用CD14抗体或p38丝裂原活化蛋白激酶(MAPK)抑制剂Mastoparan预处理PBMC后再予LPS和SAC刺激,半定量RT-PCR方法检测IL-23p19、IL-12p35和IL-12p40亚基的基因表达变化。结果:人PBMC组成性表达p19和p35,不表达p40。LPS和SAC可上调各亚基的表达。LPS诱导的IL-23和IL-12各亚基表达均需通过CDl4;CDl4仅部分参与SAC诱导的IL-12p40和p35表达上调,而与p19上调无关。LPS和SAC诱导的IL-23和IL-12各亚基表达均需要p38MAPK通路。结论:LPS和SAC刺激下IL-23和IL-12亚基表达及信号传导通路既有相似之处又有不同点,为单独或同时调控这两种因子的表达提供线索。
Objective: To evaluate the effects of lipopolysaccharide (LPS) and Staphylococcus aureus (SAC) on IL-23 and IL- 12 subunits expression in normal human peripheral blood mononuelear cells (PBMC),and explored the regulation mechanisms. Methods:PBMC were exposed to LPS or SAC, or were pretreated with CD14 antibody or p38 mitogen-acfivated protein kinase(MAPK) inhibitor Mastoparan before addition of LPS and SAC. Expression of IL-23 p19 ,IL-23 p35 and IL-12 p40 was studied by semiquantitative RTPCR. Results:IL-23 p19 and IL-12 p35 mRNAs were expressed in fresh PBMC, and expression of all IL-23 and IL-12 subunits was upregulated by LPS and SAC. LPS-induced increase of IL-23 and IL-12 subunits expression was CD14-dependent,CD14 was partially involved in SAC-induced increase of IL-12 p40 and p35 transcription,and not involved in the up-regulation of IL-23 p19 expression. Both LPS and SAC-induced subunits expression required p38 MAPK pathway. Conclusion:The expression pattern and regulation mode of IL-23 is similar to as well as distinct from that of IL-12 ,and the results may help to develop intervention strategies targeting the two cytokines either indicriminately or selectively.