CAJ | 학술논문

目的：制备粉尘螨粗抗原建立BALB/c小鼠哮喘模型，检测细胞因子并观察肥大细胞形态及脱颗粒的情况。方法：制备粉尘螨粗抗原，30只BALB/c小鼠随机均分为3组，阴性对照组（A组）、哮喘模型组（B组）和治疗组（C组）。A组始终使用PBS处理，B组和C组在第0、7和14天用50μg粉尘螨粗抗原＋50μl明矾佐剂腹腔注射致敏。在第28天，A组和B组用PBS、C组用粉尘螨粗抗原（350μg/次）皮下注射免疫1周，隔天免疫1次，末次免疫1周后开始用50μg粉尘螨粗抗原滴鼻激发，每天1次，连续7次。末次激发24h后进行气道高反应检测。末次激发48h后处死小鼠取血，对小鼠进行肺泡灌洗收集肺泡灌洗液（BALF），无菌摘取肺组织和脾脏，分别进行肺组织病理切片与脾细胞培养。检测BALF和脾细胞上清中IL-4、IL-10和IFN-γ细胞因子水平，血清中IgE、组胺抗体水平，进行HE染色和甲苯胺蓝染色观察肺部炎症细胞浸润程度和肥大细胞脱颗粒状况。结果：与B组相比，C组气道高反应性和肺部病理症状减轻（P＜0．01）。C组BALF中细胞计数结果显示与B组相比细胞总数和嗜酸粒细胞数明显减少（P＜0．01）。与B组相比，C组的肥大细胞脱颗粒情况明显减轻，细胞形态稳定。与B组（IgE：1．905）相比，C组血清中抗原特异性IgE抗体水平（IgE：1．278）明显降低（P＜0．01）。BALF细胞因子IL-4、IL-10和IFN-γ水平检测结果显示，与B组相比，C组BALF中IL-4水平明显降低接近A组值（P＜0．01），而IL-10与A、B两组相比有明显升高（P＜0．01），C组的IFN-γ也高于A、B两组（P＜0．01）。C组脾细胞上清中IL-4明显低于B组（P＜0．01），而IL-10明显高于B组（P＜0．01），IFN-γ也明显高于B组（P＜0．01）。BALF和血清中组胺水平结果显示，在BALF中，C组与B组相比略低（P＜0．05），而在血清中检测出的组胺有明显降低（P＜0．05）。结论：过敏性哮喘模型脱敏治疗后小鼠肥大细胞脱颗粒情况被抑制。目的：製備粉塵螨粗抗原建立BALB/c小鼠哮喘模型，檢測細胞因子併觀察肥大細胞形態及脫顆粒的情況。方法：製備粉塵螨粗抗原，30隻BALB/c小鼠隨機均分為3組，陰性對照組（A組）、哮喘模型組（B組）和治療組（C組）。A組始終使用PBS處理，B組和C組在第0、7和14天用50μg粉塵螨粗抗原＋50μl明礬佐劑腹腔註射緻敏。在第28天，A組和B組用PBS、C組用粉塵螨粗抗原（350μg/次）皮下註射免疫1週，隔天免疫1次，末次免疫1週後開始用50μg粉塵螨粗抗原滴鼻激髮，每天1次，連續7次。末次激髮24h後進行氣道高反應檢測。末次激髮48h後處死小鼠取血，對小鼠進行肺泡灌洗收集肺泡灌洗液（BALF），無菌摘取肺組織和脾髒，分彆進行肺組織病理切片與脾細胞培養。檢測BALF和脾細胞上清中IL-4、IL-10和IFN-γ細胞因子水平，血清中IgE、組胺抗體水平，進行HE染色和甲苯胺藍染色觀察肺部炎癥細胞浸潤程度和肥大細胞脫顆粒狀況。結果：與B組相比，C組氣道高反應性和肺部病理癥狀減輕（P＜0．01）。C組BALF中細胞計數結果顯示與B組相比細胞總數和嗜痠粒細胞數明顯減少（P＜0．01）。與B組相比，C組的肥大細胞脫顆粒情況明顯減輕，細胞形態穩定。與B組（IgE：1．905）相比，C組血清中抗原特異性IgE抗體水平（IgE：1．278）明顯降低（P＜0．01）。BALF細胞因子IL-4、IL-10和IFN-γ水平檢測結果顯示，與B組相比，C組BALF中IL-4水平明顯降低接近A組值（P＜0．01），而IL-10與A、B兩組相比有明顯升高（P＜0．01），C組的IFN-γ也高于A、B兩組（P＜0．01）。C組脾細胞上清中IL-4明顯低于B組（P＜0．01），而IL-10明顯高于B組（P＜0．01），IFN-γ也明顯高于B組（P＜0．01）。BALF和血清中組胺水平結果顯示，在BALF中，C組與B組相比略低（P＜0．05），而在血清中檢測齣的組胺有明顯降低（P＜0．05）。結論：過敏性哮喘模型脫敏治療後小鼠肥大細胞脫顆粒情況被抑製。
목적：제비분진만조항원건립BALB/c소서효천모형，검측세포인자병관찰비대세포형태급탈과립적정황。방법：제비분진만조항원，30지BALB/c소서수궤균분위3조，음성대조조（A조）、효천모형조（B조）화치료조（C조）。A조시종사용PBS처리，B조화C조재제0、7화14천용50μg분진만조항원＋50μl명반좌제복강주사치민。재제28천，A조화B조용PBS、C조용분진만조항원（350μg/차）피하주사면역1주，격천면역1차，말차면역1주후개시용50μg분진만조항원적비격발，매천1차，련속7차。말차격발24h후진행기도고반응검측。말차격발48h후처사소서취혈，대소서진행폐포관세수집폐포관세액（BALF），무균적취폐조직화비장，분별진행폐조직병리절편여비세포배양。검측BALF화비세포상청중IL-4、IL-10화IFN-γ세포인자수평，혈청중IgE、조알항체수평，진행HE염색화갑분알람염색관찰폐부염증세포침윤정도화비대세포탈과립상황。결과：여B조상비，C조기도고반응성화폐부병리증상감경（P＜0．01）。C조BALF중세포계수결과현시여B조상비세포총수화기산립세포수명현감소（P＜0．01）。여B조상비，C조적비대세포탈과립정황명현감경，세포형태은정。여B조（IgE：1．905）상비，C조혈청중항원특이성IgE항체수평（IgE：1．278）명현강저（P＜0．01）。BALF세포인자IL-4、IL-10화IFN-γ수평검측결과현시，여B조상비，C조BALF중IL-4수평명현강저접근A조치（P＜0．01），이IL-10여A、B량조상비유명현승고（P＜0．01），C조적IFN-γ야고우A、B량조（P＜0．01）。C조비세포상청중IL-4명현저우B조（P＜0．01），이IL-10명현고우B조（P＜0．01），IFN-γ야명현고우B조（P＜0．01）。BALF화혈청중조알수평결과현시，재BALF중，C조여B조상비략저（P＜0．05），이재혈청중검측출적조알유명현강저（P＜0．05）。결론：과민성효천모형탈민치료후소서비대세포탈과립정황피억제。
Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .