【摘要】 目的:探讨苦参碱抑制脂多糖(lipopolysaccharide,LPS)诱导巨噬细胞分泌炎症因子包括肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)对Toll样受体4(TLR4)、c-Jun表达的影响。方法:购买并培养小鼠RAW264.7巨噬细胞株,分成四组,即空白对照组、苦参碱组、LPS组、苦参碱干预组,通过浓度为100μg/L的LPS DMEM孵育1 h,之后将LPS弃去,然后加入不含血清的DMEM或125.25 mg/L苦参碱DMEM继续培养。分别收集上述四组细胞处理完毕后5、30、60、120min的细胞及培养液。通过PCR法检测小鼠RAW264.7巨噬细胞TLR4及c-Jun mRNA的表达,通过免疫细胞化学法检测小鼠RAW264.7巨噬细胞c-Jun蛋白的表达;通过ELISA法测定各组培养液中TNF-α、IL-1β的分泌。结果:苦参碱诱导组与空白对照组相比,TLR4 mRNA、c-Jun mRNA及c-Jun蛋白的表达量和TNF-α及IL-1β分泌量各项指标差异无统计学意义(P>0.05);LPS组各个时间点TLR4 mRNA、c-Jun mRNA及c-Jun蛋白的表达量和TNF-α及IL-1β分泌量明显高于空白对照组,且高水平能够维持2 h以上(P<0.05);苦参碱干预组能够有效的抑制LPS诱导的巨噬细胞TLR4 mRNA、c-Jun mRNA及c-Jun蛋白的表达量,降低炎症因子TNF-α、IL-1β的分泌量。结论:苦参碱可能通过抑制LPS诱导的巨噬细胞丝裂原活化的蛋白激酶信号通路上TLR4、c-Jun的过表达从而有效的降低终末炎症因子TNF-α及IL-1β的释放,进而有效的抑制炎症反应,减轻内毒素炎症反应的程度。更多还原

【Abstract】 Objective: To investigate the effect of Matrine on inhibiting lipopolysaccharide( lipopolysaccharide,LPS)-induced macrophages to secrete inflammatory cytokines,including tumor necrosis factor-α( TNF-α),leukocyte mediated element 1β( IL-1β) on Toll like receptor 4( TLR4),c-jun expression. Methods: Cultured macrophage RAW264. 7 of mouse and divided into four groups,including blank control group,matrine group,LPS group,matrine intervention group. Incubated by the concentration was 100 μg / L LPS DMEM for 1 h,then the LPS was discard. Added free serum DMEM or 125. 25 mg / L matrine DMEM to culture. The cells and culture solution of 5,30,60,120 min after the treatment of the above four groups were collected respectively. Detected of mouse RAW264. 7 macrophage TLR4 and c-jun mRNA expression by PCR. Detected of mouse macrophage RAW264. 7 cells c-jun protein expression by immunocytochemical. Measured secretion in cultured solution of TNF-α and IL-1β by ELISA. Results: Expression TLR4 mRNA,c-jun mRNA and c-jun protein and TNF-α and IL-1β secretion quantity indexes of matrine induced and blank group had no statistical significant differences( P>0. 05). TLR4 mRNA,c-jun mRNA and c-jun protein expression secretion of and TNF-α and IL-1β LPS group at each time point were significantly higher than that of blank group,and the high level to maintain more than 2 h( P<0. 05); matrine intervention group could effectively inhibit LPS induced macrophage TLR4 mRNA,c-jun mRNA and c-jun protein expression and reduce the secretion of inflammatory cytokines TNF-α and IL-1β. Conclusion: Matrine may inhibit LPS induced macrophage mitogen activated protein kinase signal pathway of TLR4 and c-jun expression so that it can effectively reduce the end inflammatory cytokines TNF-α and IL-1β release,effectively inhibit the inflammatory reaction and reduce the degree of endotoxin inflammatory response.更多还原