大黄牡丹汤含药血清对LPS刺激的小鼠肺巨噬细胞TLR4和MyD88表达的影响  

Effect of Dahuang Mudan decoction-contained serum on the expression of TLR4 and MyD88 in lipopolysaccharide-stimulated pulmonary macrophages of rats

作　　者：孙燕妮[1] 承解静[1] 杨晓燕[1] 杨洁[1] 王丽敏[1] 刘军[1] 

机构地区：[1]上海中医药大学附属普陀医院,上海200062

出　　处：《山东医药》2014年第10期10-13,共4页Shandong Medical Journal

基　　金：上海市中西结合学会基金资助项目(ZXYQ-1231)

摘　　要：目的探讨大黄牡丹汤含药血清(以下简称含药血清)对脓毒症的治疗机制。方法体外培养小鼠肺巨噬细胞株RAW264.7并随机分为空白组,模型组,含药血清高、中、低浓度组,每组3个复孔。除空白组外,其余四组均予细菌脂多糖(LPS)5μg/mL刺激24 h,含药血清高、中、低浓度组分别同时给予高、中、低浓度的含药血清进行干预。干预后于37℃、5%CO2培养箱中培养24 h。分别采用半定量RT-PCR和Western blot法测定Toll样受体4(TLR4)及髓样分化因子88(MyD88)mRNA和蛋白表达。结果模型组TLR4和MyD88 mRNA和蛋白表达均显著高于空白组(P<0.01);含药血清中浓度组与高浓度组TLR4和MyD88表达无显著差异,但均显著低于模型组(P<0.01)。结论含药血清治疗脓毒症的机制可能为抑制TLR4和下游MyD88表达。Objective To investigate the therapeutic mechanism of Dahuang Mudan decoction-contained serum （ here-inafter referred to as the drug contained serum ） in the treatment of sepsis .Methods Mouse pulmonary macrophage cell line RAW264 .7 was cultured in vitro , and then the cells were randomly divided into the blank group , model group , high-, moderate-and low-concentration drug-contained serum groups , each group of 3 holes.Except the blank group , other four groups were stimulated with bacterial lipopolysaccharide （ LPS） 5 μg/mL, then were treated with different concentrations of drug-contained serum for 24 h.After the intervention , the cells were cultured in 5%CO2 at 37℃for 24 h.Then the mR-NA expression of Toll-like receptor 4（ TLR4） and myeloid differentiation factor 88（ MyD88） was measured by semi-quanti-tative reverse transcription-polymerase chain reaction （ RT-PCR ） , and their protein expression was analyzed by Western blotting .Results The protein and mRNA expression of TLR 4 and MyD88 in the model group was significantly higher than that of the blank group （P〈0.01）, but the protein and mRNA expression of TLR4 and MyD88 in the high-and moderate-concentration drug-contained serum groups was significantly lower than that of the model group （P〈0.01）, and no signifi-cant differences was found between the high-and moderate-concentration drug-contained serum groups .Conclusion The therapeutic mechanism of Dahuang Mudan decoction-contained serum in the treatment of sepsis may be related to the inhibi-tion of TLR4 and down-regulation of MyD88 expression.

关 键 词：大黄牡丹汤 含药血清 肺巨噬细胞 TOLL样受体4 髓样分化因子88 

分 类 号：R563.9[医药卫生—呼吸系统]