摘要
目的评价迷走神经刺激对内毒素致小鼠急性肺损伤(ALI)时α7型烟碱乙酰胆碱受体(α7nAChR)信号通路与调节性T细胞(Treg细胞)的关系。方法清洁级健康雄性C57BL/6小鼠50只,6~8周龄,体重22~25g,采用随机数字表法分为5组(n=10):对照组(C组)、急性肺损伤组(ALI组)、迷走神经刺激组(VNS组)、α-银环蛇毒素组(α-BGT组)和迷走神经刺激+α-银环蛇毒素组(VNS+α-BGT组)。小鼠ALI模型制备成功后行迷走神经刺激,刺激强度0.5mA,频率20Hz,持续30s,每次电刺激脉冲间隔5min,共刺激60min。C组仅于气管内吹入等体积的无菌生理盐水,分离迷走神经但不行电刺激;ALI组制备ALI模型,分离迷走神经但不行电刺激;VNS组于ALI模型制备结束后1h行迷走神经刺激;α-BGT组腹腔注射α-银环蛇毒素1μg/kg,1h后制备ALI模型,分离迷走神经但不行电刺激;VNS+α-BGT组腹腔注射α-银环蛇毒素1μg/kg,1h后制备ALI模型,模型制备结束后1h行迷走神经刺激。于模型制备后72h,处死小鼠,取肺组织,测定肺组织含水量,流式细胞术检测Treg细胞比例,比色法检测MPO活性,Westernblot法检测α7nAChR的表达,ELISA法检测IL-10、TGF-β和IL-1β含量。结果与C组比较,余4组肺组织含水量、MPO活性和IL-1β含量升高,ALI组、α-BGT组和VNS+α-BGT组α7nAChR表达下调,VNS组Treg细胞比例升高,ALI组和α-BGT组IL-10含量降低,VNS组和VNS+α-BGT组IL-10含量升高,ALI组、α-BGT组和VNS+α-BGT组TGF-β含量降低,VNS组TGF-β含量升高(P<0.05);与ALI组比较,VNS组肺组织含水量、MPO活性和IL-1β含量降低,α7nAChR表达上调,Treg细胞比例、IL-10和TGF-β含量升高,α-BGT组TGF-β含量、VNS+α-BGT组IL-10和TGF-β含量升高(P<0.05);与VNS组比较,α-BGT组和VNS+α-BGT组肺组织含水量、MPO活性、IL-1β含量升高,α7nAChR表达下调,Treg细胞比例、IL-10含量和TGF-β含量降低(P<0.05);与α-BGT组比较,VNS+α-BGT组肺组织IL-10和TGF-β含量升高(P<0.05)。结论迷走�
Objective To evaluate the relationship between α7 nicotinic acetylcholine receptor (α7nAChR) signaling pathway and regulatory T cells (Tregs) during vagus nerve stimulation-induced reduction of endotoxin-caused acute lung injury (ALI) in mice. Methods Clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 22-25 g, were divided into 5 groups (n=10 each) using a random number table method: control group (group C), group ALI, vagus nerve stimulation group (group VNS),α-BGT group (group α-BGT) and vagus nerve stimulation plus α-BGT group (group VNS +α-BGT). After successful establishment of the model, the vagus nerve was stimulated for 30 s with a stimulus intensity of 0.5 mA, frequency of 20 Hz, an interval of 5 min, 60 min in total.Sterile normal saline 100 μl was injected into the trachea, and the vagus nerve was only exposed but not stimulated in group C. In group ALI, the ALI model was established, and the vagus nerve was isolated but not stimulated.In group α-BGT,α-BGT 1 μg/kg was intraperitoneally injected, the ALI model was established 1 h later, and the vagus nerve was isolated but not stimulated.In group VNS+α-BGT,α-BGT 1 μg/kg was intraperitoneally injected, the ALI model was established 1 h later, and the vagus nerve was stimulated at 1 h after the end of establishment.Animals were sacrificed at 72 h after establishing the model, and lungs were removed for determination of lung water content, percentage of Tregs (using flow cytometry), myeloperoxidase (MPO) activity (by colorimetric assay), expression of α7nAChR (by Western blot) and contents of interleukin-10 (IL-10), transforming growth factor-β(TGF-β) and IL-1β(by enzyme-linked immunosorbent assay). Results Compared with group C, the lung water content, MPO activity and IL-1β content were significantly increased in the other four groups, the expression of α7nAChR was significantly down-regulated in ALI,α-BGT and VNS+α-BGT groups, the percentage of Tregs was significantly increased in group VNS, the IL-10 content was signif
作者
巩红岩
郑芳
胡晓慧
刘景景
王庆志
赵国安
Gong Hongyan;Zheng Fang;Hu Xiaohui;Liu Jingjing;Wang Qingzhi;Zhao Guoan(Department of Anesthesiology, First affiliated hospital of Xinxiang Medical University,Xinxiang 453000,China;Department of Ultrasound, First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,China;School of Basic Medical Science,Xinxiang Medical University,Xinxiang 453003,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2018年第12期1513-1516,共4页
Chinese Journal of Anesthesiology
基金
河南省医学科技攻关计划联合共建项目(2018020365).
关键词
迷走神经
呼吸窘迫综合征
成人
T淋巴细胞
调节性
受体
烟碱
Vagus nerve
Respiratory distress syndrome, adult
T-Lymphocytes, regulatory
Receptors, nicotinic
