细菌溶解产物对支气管哮喘小鼠TGF-β1及Foxp3表达的影响
세균용해산물대지기관효천소서TGF-β1급Foxp3표체적영향
Impact of bacterial lysates on the expression of TGF-β1 and Foxp3 in mouse
  • 万方数据
    国际呼吸杂志 국제호흡잡지
    INTERNATIONAL JOURNAL OF RESPIRATION
    2014年 19期 1441-1446 ,共6页

    刘静%周小建%洪建国

    류정%주소건%홍건국

    支气管哮喘%细菌溶解产物%转化生长因子-β1%叉头样转录因子3 支氣管哮喘%細菌溶解產物%轉化生長因子-β1%扠頭樣轉錄因子3
    지기관효천%세균용해산물%전화생장인자-β1%차두양전록인자3
    Bronchial asthma%Broncho-Vaxom%Transforming growth factor-β1%Forkhead box P3
    目的 建立支气管哮喘(简称哮喘)小鼠模型,给予细菌溶解产物(OM-85BV)干预,通过动物实验探讨免疫调节剂-OM-85BV干预对哮喘小鼠外周血及外周免疫器官-脾脏Treg功能相关细胞因子TGF-β1及特异性转录因子Foxp3的表达的影响.方法 清洁级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:正常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天、第8天和第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天至第26天给f组小鼠OM-85BV-生理盐水(NS)溶液灌胃,其余5组以NS模拟灌胃.第27天至第31天分别给c、d、e和f组OVA-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1h给予OM-85BV干预,d组给予Dex干预,其余组用NS对照.末次激发后24 h麻醉并解剖小鼠,心脏采血测定血清TGF-β1水平,取部分右肺行HE染色作病理学检查,取胸腺组织行免疫组织化学的方法测定其中Foxp3的表达,取脾脏行Real time-PCR测调节性T细胞(Treg)相关细胞因子TGF-β1 mRNA及Foxp3 mRNA的表达.结果 与a组对照相比较,c组小鼠支气管痉挛收缩,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;c组小鼠血清中TGF-β1水平较a组显著降低,d、f组较c组情况明显增加,e组次之,差异有统计学意义(P＜0.01).胸腺组织免疫组织化学结果显示各组之间Foxp3的表达差异无统计学意义(P＞0.05),但是实验过程中可以观察到,d组胸腺体积较其他各组明显缩小.c组脾脏TGF-β1 mRNA的相对表达显著低于正常组,f组较c组表达明显增加,d组次之,差异有统计学意义(P＜0.01).c组脾脏Foxp3 mRNA的相对表达显著低于正常组,f组较c组表达明显增加,e组、d组次之,差异有统计学意义(P＜0.01).结论 口服OM-85BV干预有利于改善哮喘小鼠模型气道炎症,显著提高外周血及外周免疫器官-脾脏的Treg功能相关细胞因子TGF-β1及特异性转录因子Foxp3的表达,二者可能是预防哮喘气道炎症的发生和发展的治疗靶点.
    목적 건립지기관효천(간칭효천)소서모형,급여세균용해산물(OM-85BV)간예,통과동물실험탐토면역조절제-OM-85BV간예대효천소서외주혈급외주면역기관-비장Treg공능상관세포인자TGF-β1급특이성전록인자Foxp3적표체적영향.방법 청길급4～6주령BALB/c소서48지,수궤분위6조.a조:공백대조조;b조:정상OM-85BV대조조;c조:효천모형조;d조:지새미송(Dex)간예조;e조:OM-85BV간예A조;f조:OM-85BV간예B조(간예시간교e조연장10 d).재실험제1천、제8천화제15천분별급c、d、e、f4조소서복강주사란청백단백(OVA)-Al(OH)3현액사기치민,기여조이PBS완충액대조.제17천지제26천급f조소서OM-85BV-생리염수(NS)용액관위,기여5조이NS모의관위.제27천지제31천분별급c、d、e화f조OVA-NS용액적비격발,기여량조이등량NS대조.기중b、e화f조재격발전1h급여OM-85BV간예,d조급여Dex간예,기여조용NS대조.말차격발후24 h마취병해부소서,심장채혈측정혈청TGF-β1수평,취부분우폐행HE염색작병이학검사,취흉선조직행면역조직화학적방법측정기중Foxp3적표체,취비장행Real time-PCR측조절성T세포(Treg)상관세포인자TGF-β1 mRNA급Foxp3 mRNA적표체.결과 여a조대조상비교,c조소서지기관경련수축,지기관상피증생문란,지기관주위가견이림파세포화기산성세포위주적염증세포침윤,관강내유소량점액화탈락적상피세포,이차폐포간격증관;c조소서혈청중TGF-β1수평교a조현저강저,d、f조교c조정황명현증가,e조차지,차이유통계학의의(P＜0.01).흉선조직면역조직화학결과현시각조지간Foxp3적표체차이무통계학의의(P＞0.05),단시실험과정중가이관찰도,d조흉선체적교기타각조명현축소.c조비장TGF-β1 mRNA적상대표체현저저우정상조,f조교c조표체명현증가,d조차지,차이유통계학의의(P＜0.01).c조비장Foxp3 mRNA적상대표체현저저우정상조,f조교c조표체명현증가,e조、d조차지,차이유통계학의의(P＜0.01).결론 구복OM-85BV간예유리우개선효천소서모형기도염증,현저제고외주혈급외주면역기관-비장적Treg공능상관세포인자TGF-β1급특이성전록인자Foxp3적표체,이자가능시예방효천기도염증적발생화발전적치료파점.
    Objective To establish mouse allergic bronchial asthma (asthma) model and observe the effect of bacterial lysates (OM-85BV) on airway inflammation,as well as the expression of TGF-β1 and Foxp3 in serum and spleen.Methods Forty-eight 4 to 6 weeks healthy male BALB/c mice were used as research subjects and randomly divided into six groups,a:contral 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 by ovalbumin (OVA).Mice in groups c,d,e and f were intraperitoneally administered by 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 the 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.The TGF-β1 levels of blood serum were detected.The removed parts of lung tissue were for histological to observe airway inflammation and the thymuses were prepared for immunohistochemistry examination to detect the expression of Foxp3.The expression of TGF-β1 mRNA and Foxp3 mRNA in spleen were detected by Real time-PCR.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 evidently.TGF-β1 levels from the asthma group in blood serum were significantly lower than groups a and b while the levels of groups d and f were significantly improved than group c.The immunohistochemistry examination of Foxp3 in thymus didn' t show significant difference,but thymuses of group d were obviously smaller than other groups.Compared with the contral group (1.001±0.000),the levels of TGF-β1 mRNA from the asthma group (0.851±0.071) were significantly lower.The corresponding levels of groups d (1.085 ± 0.042) and f (1.292 ± 0.083) were significantly improved than group c (P ＜0.01).Compared with the contral group (1.001 ±0.000),the levels of Foxp3 mRNA from the asthma group (0.585 ± 0.192) were significantly lower.The corresponding levels of group f (2.074±0.037) were significantly improved than group c (P ＜0.01).Conclusions Administered by OM-85BV helps reduce airway inflammation in asthmatic mice and significantly improve the expression of TGF-β1 and Foxp3 of the peripheral blood and peripheral immune organs.The two may be the therapeutic targets to prevent the occurrence and development of airway inflammation in asthma.
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