中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2014年
4期
288-293
,共6页
鼻炎,变应性,常年性%Toll样受体4%NF-κB%脂多糖类%免疫调节
鼻炎,變應性,常年性%Toll樣受體4%NF-κB%脂多糖類%免疫調節
비염,변응성,상년성%Toll양수체4%NF-κB%지다당류%면역조절
Rhinitis,allergic,perennial%Toll-like receptor 4%NF-kappa B%Lipopolysaccharides%Immunomodulation
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.
目的 探討Toll樣受體(toll-like receptors,TLR)及覈因子κB (nuclear factor kappa B,NF-κB)信號傳導通路在變應性鼻炎(allergic rhinitis,AR)髮病中的作用,進一步研究Toll樣受體介導調節固有免疫與穫得性免疫的作用機製.方法 100隻大鼠完全隨機分成A組(對照組)、B組(AR組)、C組(AR+ LPS20組)、D組(AR+ LPS10組)、E組(AR+ LPS5組),每組20隻大鼠.B組大鼠應用卵清蛋白腹腔註射及滴鼻激髮建造AR模型,A組等量生理鹽水代替卵清蛋白,C、D、E組在卵清蛋白滴鼻激髮同時分彆加以不同濃度的脂多糖(每100μl分彆含LPS 20、10、5μg)噴鼻榦預.HE染色觀察鼻黏膜形態改變併計數炎性反應細胞浸潤數,免疫組化法檢測y榦擾素(interferon-γ,IFN-γ)、白細胞介素(interleukin,IL)4、IgE的含量,實時聚閤酶鏈反應及Western-Blot檢測TLR-4及NF-κB的錶達情況.以SPSS 13.0統計軟件進行統計學分析.結果 B組變應性損傷明顯,鼻黏膜以嗜痠粒細胞浸潤為主;IFN-γ、IL-4及IgE的錶達較A組明顯增高(P＜0.05).C、D、E組鼻黏膜以中性粒細胞浸潤為主;IFN-γ、TLR-4及NF-κB的錶達較B組增高,IL-4及IgE的錶達較B組降低,差異均有統計學意義(P值均＜0.05).其中C組TLR-4及NF-κB蛋白相對錶達量分彆為0.888 9±0.032 9、0.913 3 ±0.031 1,較A組的0.419 2±0.038 0、0.447 8±0.033 0明顯增高,較B組的0.616 1±0.025 1、0.748 1±0.034 3明顯增高,差異有統計學意義(P值均＜0.05).結論 TLR在AR髮病中髮揮瞭介導調節固有免疫與穫得性免疫的作用,使用高濃度TLR興奮劑LPS可產生免疫偏移作用,其機製可能與NF-κB過高錶達有關.
목적 탐토Toll양수체(toll-like receptors,TLR)급핵인자κB (nuclear factor kappa B,NF-κB)신호전도통로재변응성비염(allergic rhinitis,AR)발병중적작용,진일보연구Toll양수체개도조절고유면역여획득성면역적작용궤제.방법 100지대서완전수궤분성A조(대조조)、B조(AR조)、C조(AR+ LPS20조)、D조(AR+ LPS10조)、E조(AR+ LPS5조),매조20지대서.B조대서응용란청단백복강주사급적비격발건조AR모형,A조등량생리염수대체란청단백,C、D、E조재란청단백적비격발동시분별가이불동농도적지다당(매100μl분별함LPS 20、10、5μg)분비간예.HE염색관찰비점막형태개변병계수염성반응세포침윤수,면역조화법검측y간우소(interferon-γ,IFN-γ)、백세포개소(interleukin,IL)4、IgE적함량,실시취합매련반응급Western-Blot검측TLR-4급NF-κB적표체정황.이SPSS 13.0통계연건진행통계학분석.결과 B조변응성손상명현,비점막이기산립세포침윤위주;IFN-γ、IL-4급IgE적표체교A조명현증고(P＜0.05).C、D、E조비점막이중성립세포침윤위주;IFN-γ、TLR-4급NF-κB적표체교B조증고,IL-4급IgE적표체교B조강저,차이균유통계학의의(P치균＜0.05).기중C조TLR-4급NF-κB단백상대표체량분별위0.888 9±0.032 9、0.913 3 ±0.031 1,교A조적0.419 2±0.038 0、0.447 8±0.033 0명현증고,교B조적0.616 1±0.025 1、0.748 1±0.034 3명현증고,차이유통계학의의(P치균＜0.05).결론 TLR재AR발병중발휘료개도조절고유면역여획득성면역적작용,사용고농도TLR흥강제LPS가산생면역편이작용,기궤제가능여NF-κB과고표체유관.
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.
