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

摘要

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.%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。