CAJ | 학술논문

目的：观察粉尘螨粗抗原建立的 Bal b/c 哮喘模型小鼠肥大细胞的活化及细胞因子的变化。方法将20只Bal b/c小鼠按随机数字表法分为2组：阴性对照组（A组，n＝10），使用100μL PBS处理；哮喘模型组（B组，n＝10），50μg粉尘螨粗抗原＋50μL明矾佐剂分别在第0、7和14 d腹腔注射致敏1次，第28天再用50μg粉尘螨抗原滴鼻激发，每天1次，连续7次。末次激发24 h测定气道高反应；48 h处死小鼠进行眼球取血，进行肺泡灌洗并收集右肺的肺泡灌洗液（BALF），无菌取肺 HE染色病理观察、甲苯胺蓝染色观察肥大细胞脱颗粒，制取脾细胞及粉尘螨粗抗原刺激培养。采用ELISA法测定BALF和脾细胞培养上清IL-4、IL-10和IFN-γ以及血清IgE、组胺抗体含量。结果与 A组相比：B组气道高反应性和肺部病理显著加重（P＜0．01）；BALF细胞总数和嗜酸粒细胞数显著增多（P＜0．01）；肥大细胞脱颗粒现象显著加重；血清抗原特异性 IgE水平显著升高（P＜0．01）；BALF和脾细胞培养上清 IL-4和 IL-10水平显著升高（P＜0．05或P＜0．01），IFN-γ水平显著降低（P＜0．01）；BALF和血清组胺水平显著升高（P＜0．05或P＜0．01）。结论过敏性哮喘模型中肥大细胞被活化。
목적：관찰분진만조항원건립적 Bal b/c 효천모형소서비대세포적활화급세포인자적변화。방법장20지Bal b/c소서안수궤수자표법분위2조：음성대조조（A조，n＝10），사용100μL PBS처리；효천모형조（B조，n＝10），50μg분진만조항원＋50μL명반좌제분별재제0、7화14 d복강주사치민1차，제28천재용50μg분진만항원적비격발，매천1차，련속7차。말차격발24 h측정기도고반응；48 h처사소서진행안구취혈，진행폐포관세병수집우폐적폐포관세액（BALF），무균취폐 HE염색병리관찰、갑분알람염색관찰비대세포탈과립，제취비세포급분진만조항원자격배양。채용ELISA법측정BALF화비세포배양상청IL-4、IL-10화IFN-γ이급혈청IgE、조알항체함량。결과여 A조상비：B조기도고반응성화폐부병리현저가중（P＜0．01）；BALF세포총수화기산립세포수현저증다（P＜0．01）；비대세포탈과립현상현저가중；혈청항원특이성 IgE수평현저승고（P＜0．01）；BALF화비세포배양상청 IL-4화 IL-10수평현저승고（P＜0．05혹P＜0．01），IFN-γ수평현저강저（P＜0．01）；BALF화혈청조알수평현저승고（P＜0．05혹P＜0．01）。결론과민성효천모형중비대세포피활화。
Objective To observe the activation of mast cells and changes in cytokines in a Bal b/c mouse asthma model established by Dermatophagoides farinae(Der f)crude antigen.Methods Twenty Bal b/c mice were randomly divided into two groups.Mice in group A were treated with 100μL PBS.However,mice in group B were intraperitoneally injected with 50μg Der f crude an-tigen and 50μL alum adjuvant on day 0,7 and 14 for sensitization,and were intranasally chal-lenged with 50μg Der f antigen for 7 days once per day from day 28.Airway hyperresponsiveness (AHR)was determined 24 hours after the last challenge.Mice were sacrificed and blood samples were taken 48 hours after the last challenge.The right lung bronchoalveolar lavage was performed and bronchoalveolar lavage fluid(BALF)was collected.Pathological changes in the lung were ob-served by HE staining and degranulation of mast cells was analyzed by toluidine blue staining.In addition,spleen cells were separated,cultured and stimulated by Der f crude antigen.Levels of IL-4,IL-10 and IFN-γin BALF and spleen cell culture supernatant and levels of IgE and histamine antibody were determined by ELISA.Results Compared with group A,the AHR,lung inflamma-tion,total cell number in BALF,number of eosinophilic granulocytes in BALF,degranulation of mast cells,antigen-specific serum IgE amount,IL-4 and IL-10 levels in BALF and spleen cell cul-ture supernatant and histamine levels in BALF and serum significantly increased,but IFN-γlevels in BALF and spleen cell culture supernatant obviously decreased in group B(P<0.05 or P<0.01).Conclusion Mast cells were activated in the mouse allergic asthma model.