核仁素对LPS诱导的白细胞介素1β释放的影响

The effect of nucleolin on interleukin-1β secretion induced by lipopolysaccharide

目的:探讨核仁素在脂多糖(LPS)所致炎症模型中的表达及其对LPS所致的白细胞介素1β(IL-1β)释放的影响。方法:小鼠腹腔注射LPS(15mg/kg)建立内毒素血症模型,LPS(500μg/L)处理建立RAW264.7细胞炎症模型,采用免疫印迹观察核仁素在炎症模型中的表达。利用瞬时转染技术抑制或增加RAW264.7细胞内核仁素表达后,LPS处理,酶联免疫吸附实验(ELISA)检测细胞培养基中IL-1β的含量。结果:在内毒素血症小鼠的肺组织和RAW264.7细胞炎症模型中,110kD核仁素表达上调,80kD核仁素片段表达减少。与转空载体对照组比较,核仁素过表达组LPS所致的IL-1β释放明显增加(P<0.05);与正常细胞组和随机寡核苷酸组比较,核仁素低表达组LPS所致的IL-1β释放明显减少(P<0.05)。结论:在LPS所致的炎症模型中,110kD核仁素表达上调,80kD核仁素片段表达减少;核仁素促进LPS所致的IL-1β释放。

AIM： To explore the expression of nucleolin in lipopolysaccharide（ LPS）-mediated inflammatory models,and further investigate the role of nucleolin in expression and secretion of LPS-induced interleukin-1β（ IL1β） . METHODS： To establish inflammatory models,mice suffered intraperitoneal injection of LPS （ 15 mg/kg） and RAW264. 7 cells were treated with LPS（ 500 μg/L） . Western blotting were applied to identify the expression of nucleolin in these inflammatory models. After over-expression of nucleolin by transient pcDNA3. 1-C23 transfection and down-regulation by transient transfection of nucleolin antisense oligonucleotides,the secretion of IL-1β were examined by enzymelinked immunosorbent assay （ ELISA） in LPS-stimulated RAW264. 7 cells. RESULTS： Westem blotting assays showed that the 110 kD nucleolin increased in RAW264. 7 cells treated with LPS （ 500 μg/L） and the lung tissues of the mice treated with LPS （ 15 mg/kg） ,while the 80 kD component of nucleolin decreased. ELISA showed that LPS-induced IL-1β release in RAW264. 7 cells transfected with pcDNA3. 1-C23 was higher than that in pcDNA3. 1 empty vector transfected cells. LPS-induced IL-1β release in RAW264. 7 cells transfected with C23 antisense oligonucleotide was lower than that in normal cells and scramble oligonucleotide transfected cells. CONCLUSION： In LPS-mediated mouse endotoxemia model and LPS-mediated RAW264. 7 cell inflammatory model,the expression of 110 kD nucleolin was up-regulated, but 80 kD nucleolin fragment decreased. Nucleolin promoted secretion of LPS-induced IL-1β.