毒素清含药血清对脂多糖刺激下A549细胞NF-κB/MAPK信号通路的影响  

Effect of the serum containing DSQ on NF-κB/MAPK signal transduction pathways in A549 cells induced by LPS

作　　者：梅雪[1,2,3] 李建生[1,2] 田燕歌[1,2] 杨联河[3] 牛春玲[3] 

机构地区：[1]河南中医药大学呼吸疾病诊疗与新药研发河南省协同创新中心,郑州450046 [2]河南中医药大学河南省中医药防治呼吸病重点实验室,郑州450046 [3]河南中医药大学基础医学院,郑州450046

出　　处：《中药药理与临床》2017年第5期122-125,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(81202658);国家自然科学基金项目(81403367)

摘　　要：目的:探讨复方毒素清对肺上皮细胞A549内核因子κB(NF-κB)和丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)信号通路的影响,从细胞分子水平揭示毒素清治疗细菌性肺炎的作用机制。方法:LPS刺激肺泡上皮细胞A549,将培养好的A549细胞分为空白组(10%空白药物血清)、模型组(10%空白药物血清和1μg/ml LPS)、毒素清(14g/kg)组(10%含药血清和1μg/ml LPS)、毒素清(28g/kg)组(10%含药血清和1μg/ml LPS)、毒素清(56g/kg)组(10%含药血清和1μg/ml LPS),然后ELISA检测IL-1、IL-6、IL-8;实时荧光定量PCR测定TLR4mRNA和NF-κB mRNA的表达;Western Blotting检测P38MAPK、JNK46/54、ERK42/44总蛋白和磷酸化蛋白。结果:与空白组相比,模型组细胞L-1β、IL-6、IL-8、TLR4mRNA和NF-κB mRNA的表达以及P38MAPK、JNK46/54、ERK42/44蛋白的磷酸化水平均明显升高;与模型组相比,毒素清(14g/kg、28g/kg、56g/kg)的含药血清组L-1β、IL-6、IL-8、TLR4mRNA和NF-κB mRNA的表达以及P-P38MAPK、P-ERK42/44蛋白的磷酸化水平均明显降低,P-JNK46/54无显著性差异。结论:LPS能够激活肺上皮细胞A549内NF-κB和MAPK信号通路,诱导细胞因子的分泌。毒素清可通过抑制该两条信号通路,减少细胞因子的释放,从而减轻炎症反应。Objective： To investigate the influence of the serum containing Dusuqing（DSQ）,a traditional Chinese medicine prescription on NF-κB/MAPK signal transduction pathways in A549 cells induced by LPS and to reveal the mechanism of DSQ in treatment of bacterial pneumonia on the cell and molecular levels. Method： Different concentrations of the serum containing DSQ were prepared. Pulmonary epithelial cells A549 were stimulated by LPS. A549 cells were divided into five groups： control,model,serum containing DSQ（14,28,56 g/kg）. IL-1,IL-6,IL-8 were detected by enzyme-linked immunosorbent assay（ELISA）,TLR4 mRNA and NF-κB mRNA were measured by real-time fluorescent quantitative PCR（RT-PCR）,phosphorylation and total protein of P38 MAPK,JNK46/54,ERK42/44 were determined by Western blotting. Results： Significant elevation of the level of cytokines and expressions of TLRmRNA and NF-κBmRNA,and the phosphorylation of P38 MAPK,JNK46/54,ERK42/44 proteins was observed after the A549 cells were stimulated by LPS for 24 h,while the decline of them was observed in different concentrations of DSQ groups except JNK46/54. There was no difference among three different concentration groups. Conclusion：Our results demonstrate that NF-κB/MAPK pathway was involved in the A549 Cells induced by LPS,and the mechanism of DSQ onreducing inflammation reaction might be attributed to its prevention activation of NF-κB,P38,ERK pathways in the A549 Cells.

关 键 词：毒素清 脂多糖 肺上皮细胞A549 核因子κB MAPK 

分 类 号：R285.5[医药卫生—中药学]