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变应性鼻炎发病中脂多糖兴奋Toll样受体产生免疫偏移的实验研究 被引量:3

Role of immune deviation by toll-liked receptor's doping LPS in pathogenesis of allergic rhinitis
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摘要 目的 探讨Toll样受体(toll-like receptors,TLR)及核因子κB (nuclear factor kappa B,NF-κB)信号传导通路在变应性鼻炎(allergic rhinitis,AR)发病中的作用,进一步研究Toll样受体介导调节固有免疫与获得性免疫的作用机制.方法 100只大鼠完全随机分成A组(对照组)、B组(AR组)、C组(AR+ LPS20组)、D组(AR+ LPS10组)、E组(AR+ LPS5组),每组20只大鼠.B组大鼠应用卵清蛋白腹腔注射及滴鼻激发建造AR模型,A组等量生理盐水代替卵清蛋白,C、D、E组在卵清蛋白滴鼻激发同时分别加以不同浓度的脂多糖(每100μl分别含LPS 20、10、5μg)喷鼻干预.HE染色观察鼻黏膜形态改变并计数炎性反应细胞浸润数,免疫组化法检测y干扰素(interferon-γ,IFN-γ)、白细胞介素(interleukin,IL)4、IgE的含量,实时聚合酶链反应及Western-Blot检测TLR-4及NF-κB的表达情况.以SPSS 13.0统计软件进行统计学分析.结果 B组变应性损伤明显,鼻黏膜以嗜酸粒细胞浸润为主;IFN-γ、IL-4及IgE的表达较A组明显增高(P<0.05).C、D、E组鼻黏膜以中性粒细胞浸润为主;IFN-γ、TLR-4及NF-κB的表达较B组增高,IL-4及IgE的表达较B组降低,差异均有统计学意义(P值均<0.05).其中C组TLR-4及NF-κB蛋白相对表达量分别为0.888 9±0.032 9、0.913 3 ±0.031 1,较A组的0.419 2±0.038 0、0.447 8±0.033 0明显增高,较B组的0.616 1±0.025 1、0.748 1±0.034 3明显增高,差异有统计学意义(P值均<0.05).结论 TLR在AR发病中发挥了介导调节固有免疫与获得性免疫的作用,使用高浓度TLR兴奋剂LPS可产生免疫偏移作用,其机制可能与NF-κB过高表达有关. Objectives To investigate the role of TLR-NF-κB signaling pathway in pathogenesis of allergic rhinitis (AR) and the mechanism of TLR to modulate innate immunity and adaptive immunity.Methods One hundred rats were divided into 5 groups by simple randomization,normal group (group A),modle group (group B),AR + LPS20 group (group C),AR + LPS10 group (group D),AR +LPS5 group(group E).Model of AR in B group was established by intraperitoneal injection and nasal topic delivery of ovalbumin (OVA).A group was deliveried of same volume physiological saline insteated of OVA,C,D,E group were interfered by nasal delivery of LPS in different concentration (including LPS 20 μg、10 μg、5 μg per 100 μl).Changes of nasal mucosa tissues and inflammatory cell infiltration were observed by HE staining,while neutrophil and eosinophil counted under high power microscope.Expression of IL-4,IFNγ,and IgE in nasal mucosa tissues were measured with immunohistochemical method.Realtime-PCR and Western-blot were used to evaluate the expression level of TLR-4 and NF-κB in nasal mucosa tissues.SPSS 13.0 software was used to analyze the data.Results Group B was observed to have developed AR injury of nasal mucosa.Eosinophil count and the expression of IL-4,IFN-γ,and IgE were significantly higher in B group than those in A group (all P < 0.05),neutrophil count was significantly higher in C,D,E groups than that in B group (all P < 0.05).Results of immunohistochemical staining showed that,expression level of IFN-γ,TLR-4 and NF-κB were significantly higher than group B (all P < 0.05),while IL-4 and IgE were significantly decreased than group B (all P < 0.05).The protein expression of TLR-4 and NF-κB was 0.888 9 ±0.032 9 and 0.913 3 ±0.031 1 in group C,and 0.419 2 ±0.038 0 and 0.447 8 ±0.033 0 in group A,0.616 1 ± 0.025 1 and 0.748 1 ± 0.034 3 in group B,the difference was significant(all P <0.05).Conclusions TLR plays an important role of modulation between innate immunity and adaptive immunity in the pathogenesis of AR.The higher concentration of TLR doping may activate the higher expression of NF-κB then intervene the development of AR with immune deviation.
出处 《中华耳鼻咽喉头颈外科杂志》 CAS CSCD 北大核心 2014年第4期288-293,共6页 Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金 莆田市科技局课题(2010S06)
关键词 鼻炎 变应性 常年性 TOLL样受体4 NF-κB 脂多糖类 免疫调节 Rhinitis, allergic, perennial Toll-like receptor 4 NF-kappa B Lipopolysaccharides Immunomodulation
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