临床儿科杂志 ›› 2014, Vol. 32 ›› Issue (8): 767-.doi: 10.3969 j.issn.1000-3606.2014.08.018

• 实验研究 • 上一篇    下一篇

细菌溶解产物对哮喘小鼠气道炎症的预防作用

刘静,周小建,洪建国   

  1. 上海交通大学附属第一人民医院儿内科( 上海 201620)
  • 收稿日期:2014-08-15 出版日期:2014-08-15 发布日期:2014-08-15
  • 通讯作者: 洪建国 E-mail:hongjianguo@hotmail.com
  • 基金资助:
    上海市科学技术委员会重大项目(No.10D21951100); 2010 年上海市卫生局面上项目(No.2010267)

The impact of bacterial lysate on asthma prevention in mouse

LIU Jing, ZHOU Xiaojian, HONG Jianguo   

  1. Department of Pediatrics, Shanghai First People’s Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200080, China
  • Received:2014-08-15 Online:2014-08-15 Published:2014-08-15

摘要: 目的 探讨细菌溶解产物(OM-85BV)对哮喘气道炎症的预防作用及可能机制。方法 4~6周龄BALB/c小鼠48只,随机分为6组。a组,空白对照组;b组,OM-85BV对照组;c组,哮喘模型组;d组,地塞米松(Dex)干预组;e组,OM-85BV干预A组;f组,OM-85BV干预B组(干预时间较e组延长10 d)。在实验第1、8和15天分别给c、d、e、f4组小鼠腹腔注射卵清白蛋白(OVA)-Al(OH)3悬液使其致敏,其余组以PBS缓冲液对照。第17~26天,OM-85BV溶于生理盐水(NS)并给f组灌胃,其余5组以生理盐水(NS)灌胃。第27~31天分别给c、d、e和f组OVA的NS溶液滴鼻激发,其余两组以等量NS对照。其中b、e和f组在激发前给予OM-85BV灌胃,d组给予Dex灌胃,其余组用NS对照。末次激发后24 h麻醉处死小鼠,心脏采血测定血清总IgE水平,左肺行肺泡灌洗留取肺泡灌洗液(BALF)进行细胞计数并计算嗜酸性粒细胞(Eos)比例,测定BALF上清液中IFN-γ和IL-4水平,取部分右肺行苏木精-伊红(HE)染色作病理学检查。结果 与a、b组比较,c组小鼠支气管痉挛收缩明显,支气管上皮增生紊乱,支气管周围可见以淋巴细胞和嗜酸性细胞为主的炎症细胞浸润,管腔内有少量黏液和脱落的上皮细胞,而且肺泡间隔增宽;BALF中炎性细胞总数、Eos比例增高,BALF上清中IL-4水平增高,而IFN-γ水平降低,血清中总IgE水平增高,差异均有统计学意义(P<0.01);经干预的d、f组较c组情况明显改善,e组次之,以上各项指标的差异均有统计学意义(P<0.01)。结论 口服OM-85BV干预有利于调节Th1/Th2之间的平衡,减轻小鼠哮喘模型气道炎症,可有效预防支气管哮喘气道炎症的发生和发展。

Abstract: Objective To establish mouse allergic asthma model and observe the effect of bacterial lysates (OM-85BV) on airway inflammation. Methods Forty-eight 4 to 6 weeks healthy male BALB/c mice were used as research subjects and randomly divided into six groups, a: control group; b: OM-85BV contral group; c: allergic asthma model; d: dexamethasone group (Dex group); e: OM-85BV A group; f: OM-85BV B group (the intervention time was prolonged 10 days than group e). BALB/c mice were sensitized and challenged with ovalbumin (OVA). Mice in groups c, d, e and f were intraperitoneally administered with antigen (OVA)-Al(OH)3 on days 1, 8 and 15, others were administered by PBS. From the 17th day to the 26th day, Mice in group f were treated with OM-85BV and others were treated with normal saline. In the next days, mice in groups c, d, e and f were intranasal given OVA for 5 consecutive days. Additionally, mice in groups b, e and f were treated with OM-85BV before challenge, while mice in the group d were administered by Dex, others were treated with normal saline at the same dose. Twenty four hours after the last intranasal administration, mice were anesthetized and dissected. Lungs were lavaged with PBS and bronchoalveolar lavage fluid (BALF) was obtained. The total inflammatory cells and eosinophils in BALF were counted. The total IgE levels of blood serum and IFN-γ/IL-4 levels of lavage were detected. The removed parts of lung tissue were collected for histological examination. Results Compared with groups a and b, lung tissue biopsies by HE staining from the asthma group showed obvious airway inflammation. The situation of groups d and f was significantly improved than group c, while the differences between groups e and c were not evident. Total cells and the number of eosinophils in BALF of group c (90.3±13.94×104/ml) were significantly higher than that in groups a and b. Compared with the control group, levels of IL-4 in BALF (119.03±19.92 pg/ml) and IgE in serum (15.86±1.97 ng/ml) increased and levels of IFN-γ in BALF (90.50±13.51 pg/ml) reduced significantly. The corresponding levels of groups d, e and f were significantly improved than group c (P<0.01). Conclusions Administered by OM-85BV helps regulate the balance between Th1/Th2 in asthmatic mice, reduce airway inflammation, and prevent the occurrence and development of airway inflammation