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| 抑制树突状细胞的自噬改善小鼠哮喘病情 |
| 邓常文1,夏福灿2,姜岩岩3,屈玉兰1,邓捷文3,张星星1,郭振红3,白冲1* |
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(1. 第二军医大学长海医院呼吸与危重症医学科, 上海 200433;
2. 浙江大学医学院免疫学研究所, 杭州 310058;
3. 第二军医大学医学免疫学国家重点实验室, 上海 200433
*通信作者) |
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| 摘要: |
| 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、MHC Ⅱ的表达以及DCs诱导的T细胞增殖能力, 改善哮喘病情。 |
| 关键词: 树突状细胞 自噬 哮喘 细胞增殖 |
| DOI：10.16781/j.0258-879x.2016.03.0265 |
| 投稿时间：2015-11-09修订日期：2016-01-05 |
| 基金项目:国家自然科学基金(81270073, 81570019), 第二军医大学优秀硕士研究生苗子培育基金. |
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| Inhibiting autophagy of dendritic cells meliorates asthma in mice |
| DENG Chang-wen1,XIA Fu-can2,JIANG Yan-yan3,QU Yu-lan1,DENG Jie-wen3,ZHANG Xing-xing1,GUO Zhen-hong3,BAI Chong1* |
(1. Department of Respiratory and Critical Care Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou 310058, Zhejiang, China;
3. National Key Laboratory of Medical Immunology, Second Military Medical University, Shanghai 200433, China
*Corresponding author) |
| Abstract: |
| Objective To study the impact of autophagy inhibition on functions of dendritic cells (DCs) and mouse model of allergic asthma. Methods (1)BALB/c mice were divided into three groups using the random number table: asthma model group, asthma treated with autophagy inhibitor (chloroquine) group (asthma+CQ group) and control group. Mouse models in asthma group and asthma+CQ group were induced with ovalbumin (OVA); meanwhile, mice of asthma+CQ group were also treated with autophagy inhibitor CQ. Hematoxylin-Eosin (H-E) staining was used to observe the pathological changes in lung tissues of mice. ELISA, Western blotting analysis and flow cytometery were used to detect the serum OVA-specific IgE, autophagy level, and expression of surface co-stimulatory molecules and major histocompatibility complex class Ⅱ(MHC Ⅱ) on lung DCs, respectively. (2)Bone marrow-derived DCs were treated with autophagy inhibitor 3-methyladenine (3MA) in vitro and the surface expression of co-stimulatory molecules and MHC Ⅱ was detected. (3) We sorted CD4+ T cells from spleens of OT2 mice, then co-cultured with lung DCs from mice of different groups (T cells:DCs=1:10), and detected the activation and proliferation of T cells with flow cytometery. Results (1) The level of OVA-specific IgE (P<0.05), extent of inflammatory cell infiltration in lung tissues, autophagy level in lung DCs (P<0.05), and expression of CD86 and MHCⅡ (P<0.05)on lung DCs in asthma+CQ group were significantly lower than those in the asthma group. (2) 3-MA treatment decreased the surface expression of CD86 and MHCⅡ on bone marrow-derived DCs (P<0.05). And (3) lung DCs from asthma+CQ group had lower ability for activating T cells and promoting T cell proliferation than those from the asthma group (P<0.05). Conclusion Autophagy inhibitors can improve the pathologic condition of allergic asthma through inhibiting autophagy in DCs, down-regulating surface expression of co-stimulatory molecules and MHCⅡon DCs, and further inhibiting the DCs-induced proliferation of T cells. |
| Key words: dendritic cells autophagy asthma cell proliferation |
