槐定碱对小鼠巨噬细胞TLR4及JNK/c-jun mRNA表达的影响  

Effects of Sophoridine on the Expression of TLR4 and JNK/c-jun mRNA in Mouse Macrophage

作　　者：王艳荣[1] 刘静 摆茹[1] 梁锦屏[1] 赵建宁[1] 王宁萍[1] 周娅[1] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与免疫学系,银川750004 [2]湖北省咸宁市中心医院检验科,咸宁437100

出　　处：《宁夏医科大学学报》2011年第4期315-317,355,共4页Journal of Ningxia Medical University

基　　金：国家自然科学基金(30660227);宁夏回族自治区自然科学基金(NZ0783);宁夏回族自治区卫生厅科研项目(2008022)

摘　　要：目的观察槐定碱预处理对LPS刺激RAW 264.7巨噬细胞表达TLR4、JNK和c-jun mRNA及分泌TNF-α的影响,探讨槐定碱抗内毒素机制。方法培养RAW 264.7巨噬细胞,实验分为4组:空白对照组、槐定碱对照组、LPS组、槐定碱预处理组,用RT-PCR技术检测RAW 264.7细胞TLR4、JNK和c-jun的mR-NA表达量,放免法检测细胞培养液TNF-α分泌量。结果槐定碱对RAW 264.7巨噬细胞TLR4、JNK、c-jun的mRNA及细胞培养液中TNF-α基础表达无影响,但对经LPS激活的巨噬细胞的TLR4、JNK mRNA表达有显著抑制作用(均P<0.01),c-jun mRNA表达亦降低,但与LPS模型组比较差异尚无统计学意义。槐定碱预处理组细胞液中TNF-α含量显著低于LPS模型组(P<0.01)。结论槐定碱抗内毒素机制与其抑制LPS引起的TLR4、JNK的mRNA高表达,并减少TNF-α分泌有关。Objective To explore the anti-endotoxin mechanisms by observing the effects of Sophoridine pretreating on the expression of TLR4 and its downstream signaling molecule JNK/c-jun mRNA and secretory of TNF-α in macrophage induced by LPS.Methods RAW264.7 cells were cultured and divided into 4 groups： macrophage control group,sophoridine control group,LPS model group,Sophoridine pretreating group.The expression of TLR4 and JNK/c-jun mRNA in RAW264.7 cells were detected by reverse transcription polymerase chain reaction（RT-PCR）,and the secretory volume of TNF-α in cell culture fluid were determined by radioimmunoassay.Results Sophoridine had no effect on the basic expression of TLR4 and JNK/c-jun mRNA in RAW264.7 cells.Sophoridine had inhibition on the expression of TLR4 and JNK mRNA induced by LPS（all P0.01） but no significant difference was found on the inhibited expression of c-jun mRNA between sophoridine pretreating group and LPS group.The results showed that the secretory volume of TNF-α in cell culture fluid of sophoridine pretreating group was lower than that in LPS group（P0.01）.Conclusion The anti-endotoxin mechanisms of Sophoridine is related with inhibiting the high expression of TLR and JNK mRNA and reducing secretory volume of TNF-α induced by LPS.

关 键 词：槐定碱 脂多糖 TLR4 JNK C-JUN 

分 类 号：R285.5[医药卫生—中药学]