本文選題：哮喘 + 動物模型��； 參考：《遵義醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的：評價以雞卵清白蛋白(OVA)致敏和激發(fā)小鼠制備哮喘模型。 方法：清潔級BALB/c小鼠20只隨機分為哮喘組(AS組)和生理鹽水對照組(NS組),每組10只。AS組小鼠于第1、13d以O(shè)VA致敏,從第1次致敏后第19d開始以5%OVA溶液霧化激發(fā)并連續(xù)5d,末次激發(fā)24h后處死小鼠；NS組以生理鹽水代替OVA作對照,余處理同AS組。檢測兩組小鼠支氣管肺泡灌洗液(BALF)中細胞總數(shù)和細胞分類計數(shù),采用酶聯(lián)免疫吸附試驗(ELISA)檢測BALF白細胞介素5(IL-5)和IL-32水平的變化,蘇木素-伊紅(HE)染色觀察肺組織病理學(xué)改變,阿辛藍-過碘酸雪夫(AB-PAS)染色觀察氣道上皮杯狀細胞及黏液物質(zhì)改變,Van Gieson(VG)染色觀察氣道周圍膠原纖維增生情況,免疫組織化學(xué)(IHC)檢測肺組織中黏蛋白5ac(Muc5ac)和IL-32蛋白的表達水平,實時熒光定量逆轉(zhuǎn)錄PCR和免疫印跡法(Western blot)分別檢測肺組織中Muc5ac mRNA和蛋白的表達水平。 結(jié)果：AS組小鼠肺組織中出現(xiàn)哮喘特征性病理改變,BALF中細胞總數(shù)、嗜酸性粒細胞、淋巴細胞、中性粒細胞百分比、IL-5和IL-32水平、氣道炎癥病理評分、氣道上皮杯狀細胞及黏液物質(zhì)陽性相對著色面積、氣道周圍膠原纖維陽性相對著色面積、肺組織IL-32蛋白、Muc5ac蛋白和Muc5ac mRNA表達均高于NS組,差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論：以O(shè)VA致敏和激發(fā)小鼠可以成功復(fù)制哮喘小鼠動物模型；哮喘小鼠出現(xiàn)明顯的氣道炎癥、氣道上皮杯狀細胞增生／化生及氣道黏液高分泌；哮喘小鼠氣道炎癥可能導(dǎo)致氣道黏液高分泌的發(fā)生 目的：通過應(yīng)用羅格列酮和地塞米松干預(yù)哮喘小鼠,探討胸腺基質(zhì)淋巴細胞生成素(TSLP)在哮喘小鼠氣道黏液高分泌中的作用。 方法：清潔級BALB/c小鼠40只隨機分成生理鹽水對照組(NS組)、哮喘組(AS組)、羅格列酮干預(yù)組(AS+ROS組)、地塞米松干預(yù)組(AS+DEX組),各10只。AS組和NS組處理同第一部分；AS+ROS組從第1次致敏后第17d開始予羅格列酮5mg*kg-1*d-1灌胃,第19d開始,于每日霧化激發(fā)前30min給藥,連續(xù)7d,余處理同AS組；AS+DEX組于每日霧化激發(fā)前30min予地塞米松2mg/kg腹腔注射,連續(xù)5d,余處理同AS組。測定各組小鼠BALF中細胞總數(shù)和細胞分類計數(shù),采用ELISA檢測BALF中IL-5水平的變化,HE染色觀察肺組織病理學(xué)改變,AB-PAS染色觀察氣道上皮杯狀細胞及黏液物質(zhì)改變,VG染色觀察氣道周圍膠原纖維增生情況,IHC檢測肺組織中Muc5ac、TSLP、CD11。和脾組織中CD11。蛋白的表達水平,實時熒光定量逆轉(zhuǎn)錄PCR和Western blot分別檢測肺組織中Muc5ac、TSLP mRNA和蛋白的表達水平。 結(jié)果：AS組BALF細胞總數(shù)、嗜酸性粒細胞、淋巴細胞、中性粒細胞百分比、IL-5水平、氣道炎癥病理評分、氣道上皮杯狀細胞及黏液物質(zhì)陽性相對著色面積、氣道周圍膠原纖維陽性相對著色面積、肺組織和脾組織中CD11C蛋白、肺組織中Muc5ac、 TSLP蛋白及Muc5ac、TSLP mRNA表達均較NS組、AS+ROS組、AS+DEX組增高,差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論：哮喘小鼠氣道上皮細胞中TSLP表達增高,TSLP可能通過激活CD11c+DCs誘導(dǎo)CD4+T細胞前體向Th2細胞分化,啟動Th2細胞優(yōu)勢免疫應(yīng)答,加重哮喘氣道炎癥,導(dǎo)致氣道Muc5ac mRNA和蛋白的高表達,促進氣道黏液高分泌；羅格列酮和地塞米松可能通過抑制肺組織中TSLP勺表達,減少Th2細胞因子的分泌,從而減輕哮喘小鼠的氣道炎癥和氣道黏液高分泌。
[Abstract]:Objective : To evaluate the effect of OVA on the preparation of asthma model .

Methods : 20 BALB / c mice were randomly divided into two groups : asthma group ( AS group ) and normal saline control group ( NS group ) . Each group of 10 . AS group mice were sensitized with OVA on Days 1 and 13d . After the first sensitization , the mice were killed at 5 % OVA solution for 5 days , and the mice were sacrificed at the last 24 hours ;
The expression levels of Muc5ac ( Muc5ac ) and IL - 32 in lung tissues were observed by enzyme - linked immunosorbent assay ( ELISA ) , and the expression level of Muc5ac ( Muc5ac ) and IL - 32 protein in lung tissues were detected by immunohistochemistry ( IHC ) , and the level of Muc5ac mRNA and protein expression in lung tissues was detected by real - time fluorescence quantitative reverse transcription - PCR and Western blot .

Results : The expression of IL - 32 , Muc5ac and Muc5ac mRNA in BALF were higher than that in NS group ( P0.05 ) .

Conclusion : OVA - sensitized and challenged mice can successfully replicate the mouse model of asthma mice .
In asthmatic mice , airway inflammation , airway epithelial cup - like cell proliferation / chemical and airway mucus hypersecretion were observed .
Airway inflammation in asthmatic mice may cause airway mucus hypersecretion

Objective : To study the role of thymic stromal lymphopoiesis ( TSLP ) in the airway mucus hypersecretion of asthmatic mice by the intervention of roglione and dexamethasone in the treatment of asthma mice .

鏂規(guī)硶錛氭竻媧佺駭BALB/c灝忛紶40鍙殢鏈哄垎鎴愮敓鐞嗙洂姘村鐓х粍(NS緇，

本文編號：1972805