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[目的]观察大承气汤(DD)含药血清对人支气管上皮细胞(HBE)表达Toll样受体4(TLR-4)的影响,探讨DD对脂多糖(LPS)刺激的肺损伤的保护作用及机制。[方法]制备DD及DD含药血清。将体外培养的HBE分为7组:正常血清对照组,单纯LPS干预组,低剂量DD含药血清组,中剂量DD含药血清组,高剂量DD含药血清组,西药对照组,空白血清对照组。采用免疫细胞化学法、Real-time PCR及Western blot检测不同剂量的DD含药血清对LPS刺激的HBE细胞表达TLR-4mRNA及蛋白的影响。[结果]单纯LPS干预组TLR-4蛋白表达水平较正常血清对照组显著增加(P<0.01),高、中、低剂量DD含药血清组TLR-4蛋白表达水平较单纯LPS干预组下降。[结论]DD含药血清能抑制LPS刺激的HBE表达TLR-4,这可能是DD对肠源性肺损伤具有保护作用的机制之一。
[Objective] To observe the effect of Dachengqi Decoction (DD) containing serum on Toll-like receptor 4 (TLR-4) expression in human bronchial epithelial cells (HBE), and to explore the protective effect of DD on lipopolysaccharide (LPS) -induced lung injury Role and mechanism. [Method] DD and DD drug-containing serum were prepared. HBE cultured in vitro were divided into 7 groups: normal serum control group, simple LPS intervention group, low dose DD serum-containing group, medium dose DD serum-containing group, high dose DD serum-containing group, western medicine control group, blank serum Control group. Immunocytochemistry, Real-time PCR and Western blot were used to detect the effects of different doses of DD-containing serum on TLR-4 mRNA and protein expression in LPS-stimulated HBE cells. [Results] The expression of TLR-4 protein in LPS intervention group was significantly higher than that in normal serum control group (P <0.01). The expression of TLR-4 protein in serum of high, medium and low dose DD group was lower than that of LPS intervention group. [Conclusion] DD-containing serum can inhibit LPS-stimulated expression of TLR-4 in HBE, which may be one of the mechanisms by which DD can protect gut-derived lung injury.