巨噬细胞中脂多糖诱导微小RNA-155表达与地塞米松的抑制

Dexamethasone inhibits the expression of micro RNA-155 in macrophages induced by lipopolysaccharide

背景:目前地塞米松对巨噬细胞中微小RNA-155表达的调控作用尚不明确。目的:了解地塞米松是否调节巨噬细胞中微小RNA-155的表达。方法:1脂多糖刺激小鼠巨噬细胞:体外培养小鼠巨噬细胞株Raw264.7细胞,予脂多糖刺激。分别在培养0,0.5,2,6 h收集细胞,检测miR NA-155的动态表达。2地塞米松对巨噬细胞的干预:实验分4组:对照组予磷酸盐缓冲液培养;脂多糖组予脂多糖刺激;地塞米松+脂多糖组予地塞米松和脂多糖共同作用;地塞米松组予地塞米松培养。6 h后收集培养上清用ELISA法检测培养液中肿瘤坏死因子α、白细胞介素6等炎症因子表达,用实时荧光定量PCR法检测巨噬细胞中微小RNA-155的表达。结果与结论:1脂多糖刺激Raw264.7巨噬细胞6 h后炎症因子肿瘤坏死因子α、白细胞介素6及微小RNA-155表达明显增加(P<0.05)。2用地塞米松+脂多糖干预后炎症因子及微小RNA-155的表达轻度升高(P<0.05)。3单独地塞米松处理组中炎症因子无明显变化(P>0.05),但微小RNA-155却明显减低(P<0.05)。4结果说明地塞米松抑制脂多糖诱导的巨噬细胞中微小RNA-155的表达。

BACKGROUND： It is unclear about dexamethasone effect on the regulation of micro RNA-155 expression in macrophages.OBJCTIVE： To explore whether dexamethasone can regulate the expression of micro RNA-155 in macrophages. METHODS：（1） Lipopolysaccharide stimulation of mouse macrophages： mouse macrophage cell lines, Raw264.7 cells, were cultured in vitro and stimulated by lipopolysaccharide. Cultured cells were collected at 0, 0.5, 2, 6 hours after culture to detect the dynamical expression of microR NA-155.（2） Dexamethasone intervention for macrophages： Macrophages were divided into four groups： control group treated with phosphate buffer; lipopolysaccharide group stimulated by lipopolysaccharide; combined group given intervention with dexamethasone and lipopolysaccharide; dexamethasone group cultured with dexamethasone. At 6 hours after culture, cell supernatant was collected to detect the expression of tumor necrosis factor α and interleukin-6 using ELISA method. Real-time fluorescence quantitative PCR was used to detect the expression of microR NA-155 in the Raw264.7 macrophages. RESULTS AND CONCLUSION： Lipopolysaccharide significantly increased the expression of tumor necrosis factor α, interleukin-6 and microR NA-155 after 6 hours of culture（P〈0.05）. Combined use of dexamethasone and lipopolysaccharide slightly increased the expression of tumor necrosis factor α, interleukin-6 and micro RNA-155（P〈0.05）. Dexamethasone alone had no influence on the expression of tumor necrosis factor α and interleukin-6, but significantly decreased the expression of micro RNA-155（P〈0.05）. These findings indicate that dexamethasone can inhibit the expression of micro RNA-155 in the macrophages induced by lipopolysaccharide.