丙泊酚调控巨噬细胞极化对支气管哮喘小鼠气道炎症反应和Toll样受体4-NOD样受体蛋白3通路的影响

Effect of propofol regulating macrophage polarization on airway inflammatory response and Toll-like receptor 4-NOD-like receptor protein 3 pathway in mice with bronchial asthma

  • 摘要:
    目的 探讨丙泊酚(Pro)调控巨噬细胞极化对支气管哮喘(BA)小鼠气道炎症反应和Toll样受体4-NOD样受体蛋白3(TLR4-NLRP3)通路的影响。
    方法 将40只BA造模小鼠随机分为BA组、Pro低剂量(Pro-L)组、Pro高剂量(Pro-H)组、Pro-H+脂多糖(LPS)组,每组10只。另取10只正常小鼠作为Control组。检测各组小鼠肺功能最大呼气流量(PEF)、每分钟通气量(VE), 肺泡灌洗液中嗜酸性粒细胞(EOS)、淋巴细胞(LYM)和中性粒细胞(NEU)数量,白细胞介素(IL)-4、IL-10、IL-5和IL-13水平; 采用流式细胞术检测外周血巨噬细胞M1型和M2型水平,辅助性T细胞(Th)1和Th2细胞比例; 采用酶联免疫吸附测定(ELISA)检测血清γ干扰素(IFN-γ)、免疫球蛋白E(IgE)水平; 采用苏木精-伊红(HE)染色观察肺组织病理形态; 采用Western blot检测肺组织中cleaved caspase-3、TLR4、NLRP3和Caspase-1蛋白表达。
    结果 与Control组比较, BA组小鼠肺组织有明显损伤, PEF、VE、IL-10、巨噬细胞M1型、Th1细胞比例、IFN-γ水平降低, EOS、LYM、NEU数量以及IL-4、IL-5、IL-13、巨噬细胞M2型水平、Th2细胞比例、IgE、cleaved caspase-3、TLR4、NLRP3和Caspase-1蛋白表达水平升高,差异有统计学意义(P < 0.05); 与BA组相比, Pro-L组、Pro-H组小鼠肺组织有明显损伤,PEF、VE、IL-10、巨噬细胞M1型水平、Th1细胞比例、IFN-γ水平升高, EOS、LYM、NEU数量以及IL-4、IL-5、IL-13水平、巨噬细胞M2型水平、Th2细胞比例、IgE、cleaved caspase-3、TLR4、NLRP3和Caspase-1蛋白表达水平降低,差异有统计学意义(P < 0.05); LPS可显著减弱Pro对BA小鼠的改善作用(P < 0.05)。
    结论 Pro可能通过抑制TLR4-NLRP3信号通路来调节BA小鼠巨噬细胞极化和免疫反应,降低炎症反应程度,改善肺组织形态和肺功能。

     

    Abstract:
    Objective To investigate the effect of propofol (Pro) regulating macrophage polarization on airway inflammatory response and Toll-like receptor 4-NOD-like receptor protein 3 (TLR4-NLRP3) pathway in mice with bronchial asthma (BA).
    Methods Forty BA model mice were randomly divided into BA group, low-dose Pro (Pro-L) group, high-dose Pro (Pro-H) group, and Pro-H+lipopolysaccharide (LPS) group, with 10 mice in each group. Additionally, 10 normal mice were included as control group. Lung function indicatorspeak expiratory flow (PEF) and ventilation volume (VE), eosinophil (EOS), lymphocyte (LYM) and neutrophil (NEU) counts in bronchoalveolar lavage fluid, and interleukin (IL)-4, IL-10, IL-5 and IL-13 levels were measured in each group; flow cytometry was used to detect M1 and M2 macrophage levels and the proportions of T helper (Th) 1 and Th2 cells in peripheral blood; the enzyme-linked immunosorbent assay (ELISA) was used to detect serum interferon-γ (IFN-γ) and immunoglobulin E (IgE) levels; the hematoxylin-eosin (HE) staining was used to observe the pathological morphology of lung tissues; the Western blot was used to detect the protein expression of cleaved caspase-3, TLR4, NLRP3 and Caspase-1 in lung tissues.
    Results Compared with the control group, mice in the BA group showed significant lung tissue damage, decreased PEF, VE, IL-10 and M1 macrophage levels, Th1 cell proportion, and IFN-γ level, and significant increased EOS, LYM, NEU counts, IL-4, IL-5, IL-13 and M2 macrophage levels, Th2 cell proportion, IgE, cleaved caspase-3, TLR4, NLRP3, and Caspase-1 protein expression levels (P < 0.05). Compared with the BA group, mice in the Pro-L and Pro-H groups showed significant lung tissue damage, increased PEF, VE, IL-10 and M1 macrophage levels, Th1 cell proportion, and IFN-γ level, and significant decreased EOS, LYM, NEU counts, IL-4, IL-5, IL-13 and M2 macrophage levels, Th2 cell proportion, IgE, cleaved caspase-3, TLR4, NLRP3, and Caspase-1 protein expression levels (P < 0.05). LPS significantly attenuated the improvement effect of Pro in BA mice (P < 0.05).
    Conclusion Pro may regulate macrophage polarization and immune response in BA mice by inhibiting the TLR4-NLRP3 signaling pathway, reducing the degree of inflammatory response, and improving lung tissue morphology and lung function.

     

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