S100A8对大鼠肺泡巨噬细胞炎性因子释放水平的影响

Effect of S100A8 on the release of inflammatory factors from rat alveolar macrophages

目的:探讨S100A8 siRNA对脂多糖诱导的大鼠肺泡巨噬细胞NR8383炎症介质释放水平的影响。方法:构建S100A8 siRNA载体,利用脂质体Lipofectamine^(TM)2000转染NR8383细胞,荧光定量PCR检测S100A8基因的沉默效率。不同浓度(0.1~10μg/mL)的脂多糖刺激S100A8沉默组和阴性对照组细胞2 h,酶联免疫吸附法(ELISA)检测上清液中白细胞介素-6(IL-6)、IL-8和肿瘤坏死因子-α(TNF-α)的浓度。结果:S100A8 siRNA转染至NR8383细胞后,S100A8 mRNA的表达降低(P<0.05)。随着脂多糖刺激浓度增加,S100A8沉默组和阴性对照组细胞上清液中IL-6、IL-8和TNF-α浓度均逐渐增加,各浓度组与0μg/mL组比较,差异有统计学意义(P<0.05);各浓度组之间两两比较,差异均有统计学意义(P<0.05)。以相同浓度脂多糖刺激S100A8沉默组和阴性对照组细胞,阴性对照组细胞上清液中IL-6、IL-8和TNF-α浓度均高于S100A8沉默组(P<0.05)。结论:S100A8 siRNA通过下调NR8383细胞中S100A8基因的表达,减轻了脂多糖诱导的炎症介质的释放,有望作为炎症疾病治疗的潜在靶点。

Objective:To explore the effect of S100A8 siRNA on the release of inflammatory mediators from lipopolysaccharide(LPS)induced rat alveolar macrophages NR8383.Methods:S100A8 siRNA was constructed and transfected into NR8383 cells with Lipofectamine^(TM)2000.The silencing efficiency of S100A8 gene was detected by RT-qPCR.The cells of S100A8 silence group and negative control group were stimulated with different concentrations of LPS(0.1μg/ml-10μg/ml)for 2 hours,and enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IL-6,IL-8 and TNF-αin the supernatant.Results:After S100A8 siRNA was transfected into NR8383 cell,the expression of S100A8 mRNA was significantly reduced(P<0.05).As the increase of LPS stimulation concentration,the concentrations of IL-6,IL-8 and TNF-αin the supernatant of S100A8 silencing group and negative control group gradually increased,there was significant difference between concentration groups and 0μg/mL group(P<0.05).There was significant difference between each concentration groups(P<0.05).Used the same concentration of LPS to stimulate the S100A8 silenced-cells and the negative control-cells,the concentrations of IL-6,IL-8 and TNF-αin the supernatant of the negative control group were higher than those of the S100A8 silenced group(P<0.05).Conclusion:S100A8 siRNA reduces the release of inflammatory mediators induced by lipopolysaccharide by down-regulating the expression of S100A8 gene in NR8383 cells,and is expected to be a potential target for the treatment of inflammatory diseases.