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目的研究抑制树突状细胞(DCs)自噬对DCs功能及小鼠哮喘的影响。方法 (1)将BALB/c小鼠按随机数字表法分为3组:急性哮喘对照组(哮喘组)、哮喘自噬抑制剂氯喹(CQ)治疗组(CQ组)和空白对照组(对照组)。哮喘组和CQ组利用卵清蛋白(OVA)诱导建立小鼠过敏性哮喘模型,CQ组加用CQ。用H-E染色观察各组小鼠肺组织的病理改变,酶联免疫吸附实验、蛋白质印迹实验、流式细胞术分别检测各组小鼠血清OVA特异性IgE以及肺DCs自噬水平、表面共刺激分子和主要组织相容性复合体Ⅱ类分子(MHCⅡ)的表达水平。(2)体外用自噬抑制剂3-甲基腺嘌呤(3MA)抑制C57/B6小鼠骨髓源DCs自噬,检测DCs表面共刺激分子、MHCⅡ的表达水平。(3)获取OT2小鼠脾脏CD4+T淋巴细胞,与各组肺DCs以1∶10的比例混匀,流式细胞术检测T细胞的增殖情况与活化反应。结果 (1)CQ组IgE的表达水平(P<0.05)、肺炎症细胞浸润程度以及肺DCs自噬水平(P<0.05)和CD86、MHCⅡ表达水平(P<0.05)均低于哮喘组。(2)3MA体外抑制骨髓源DCs自噬后,DCs表面的CD86及MHCⅡ表达均低于哮喘组(P<0.05)。(3)CQ组肺DCs体外诱导的T细胞增殖能力与活化反应均弱于哮喘组(P<0.05)。结论自噬抑制剂可抑制过敏性哮喘肺DCs的自噬,下调DCs表面共刺激分子、MHCⅡ的表达以及DCs诱导的T细胞增殖能力,改善哮喘病情。
Objective To study the effect of dendritic cells (DCs) autophagy on DCs function and asthma in mice. Methods (1) BALB / c mice were divided into 3 groups according to random number table: acute asthma control group (asthma group), CQ group (CQ group) and blank control group group). Allergic asthma model was induced by ovalbumin (OVA) in asthma group and CQ group, and CQ was used in CQ group. The pathological changes of the lung tissues of each group were observed by HE staining. The levels of serum OVA-specific IgE and the level of autophagy of lung DCs were detected by enzyme-linked immunosorbent assay, Western blotting and flow cytometry. The surface costimulatory molecules And major histocompatibility complex class II molecules (MHC II) expression levels. (2) In vitro autophagy inhibition of 3-methyladenine (3MA) on bone marrow-derived DCs in C57 / B6 mice was used to detect the expression of co-stimulatory molecules and MHCⅡ on DCs. (3) Obtaining spleen CD4 + T lymphocytes of OT2 mice, mixing them with lung DCs of each group at a ratio of 1:10, measuring the proliferation and activation of T cells by flow cytometry. Results (1) The expression of IgE in CQ group was significantly lower than that in asthma group (P <0.05). The infiltration of inflammatory cells in lung tissue and autophagy of DCs (P <0.05) and CD86 and MHCⅡ expression in lung were lower than those in asthma group. (2) After 3MA inhibited the autophagy of bone marrow-derived DCs in vitro, the expressions of CD86 and MHCⅡ on DCs were lower than those in asthma group (P <0.05). (3) The proliferation and activation of T cells induced by DC in vitro in CQ group were weaker than those in asthma group (P <0.05). Conclusion The autophagy inhibitor can inhibit the autophagy of allergic asthmatic lung DCs, downregulate the expression of co-stimulatory molecules on surface of DCs, the expression of MHC Ⅱ and the proliferation of T cells induced by DCs, and improve the condition of asthma.