【摘要】 体外培养小鼠巨噬细胞RAW264.7,实验分为空白组、脂多糖(LPS)组(1mg/LLPS)及脂氧素A4(LXA4)处理组(1mg/LLPS与0.1~1000nmol/LLXA4共同温育)。处理预设时间后,实时荧光定量聚合酶链反应和酶联免疫吸附实验分别检测粒细胞集落刺激因子(G-CSF)基因表达和蛋白质分泌水平,免疫印迹法检测IκBα降解和NF-κB转位情况,荧光素酶报告质粒检测NF-κB转录活性。结果表明:LPS诱导G-CSF表达(P<0.01),LXA4抑制G-CSF基因表达和蛋白质分泌,其中10nmol/LLXA4作用最明显(抑制率为39.8%)(P<0.01);10nmol/LLXA4明显抑制IκBα降解(P<0.05)、NF-κB转位(P<0.05)以及NF-κB的转录活性(P<0.05)。这提示LXA4可能通过抑制NF-κB的转位和转录活性而抑制LPS诱导RAW264.7巨噬细胞分泌G-CSF。更多还原

【Abstract】 To explore the effects of lipoxin A4 (LXA4) on granulocyte colony stimulating factor (G-CSF) expression in RAW264.7 macrophages and the possible mechanisms. After treated with 1 mg/L lipopolysaccharide (LPS) in the absence or presence of LXA4 (0.1–1 000 nmol/L), the cells were harvested and G-CSF gene expression levels were assessed by real time PCR, G-CSF protein concentrations were determined by ELISA, IκBα degradation and NF-κB translocation were detected through Western blot, NF-κB transcriptional activities were tested by trans-fections and luciferase activities assay. Results demonstrated that LPS increased G-CSF gene and protein expres-sion levels, LXA4 inhibited the effects of LPS, and 10 nmol/L LXA4 restrained LPS-induced IκBα degradation, NF-κB translocation and NF-κB transcriptional activity. In a word, LXA4 controlled LPS-induced G-CSF production through inhibition NF-κB activity in RAW264.7 macrophages.更多还原