本文選題：哮喘　切入點(diǎn)：小鼠　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的:通過(guò)雞卵清白蛋白(OVA)致敏的哮喘小鼠模型,觀察羧甲司坦干預(yù)對(duì)哮喘小鼠轉(zhuǎn)化生長(zhǎng)因子-β1介導(dǎo)的氣道重構(gòu)的影響。方法:將32只BALB/c雌性小鼠在正常環(huán)境下飼養(yǎng)1周后,采用隨機(jī)法將小鼠分為正常對(duì)照組(NS)、哮喘模型組(AS)、羧甲司坦組(AS+SCMC)、地塞米松組(AS+DEX)4組,每組8只。哮喘組、羧甲司坦干預(yù)組、地塞米松干預(yù)組于第1、13天分別以O(shè)VA腹腔聯(lián)合皮下注射致敏,第19d開(kāi)始予10%OVA溶液連續(xù)霧化激發(fā)18d,末次激發(fā)24h后取材,每次激發(fā)前30min AS+SCMC組予羧甲司坦(10mg/kg)灌胃處理,AS+DEX組予以地塞米松(2mg/kg)腹腔注射,AS組以生理鹽水代替。NS組霧化及處理全部以生理鹽水代替。測(cè)定各組小鼠BALF細(xì)胞總數(shù)和細(xì)胞分類(lèi)計(jì)數(shù),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(xì)胞和黏液物質(zhì)改變,MASSON染色觀察氣道周?chē)z原纖維沉積,ELISA檢測(cè)BALF細(xì)胞因子TGF-β1的水平,免疫組化染色和實(shí)時(shí)熒光定量PCR分別檢測(cè)肺組織TGF-β1蛋白和m RNA表達(dá)水平。結(jié)果:AS+SCMC組、AS+DEX組BALF細(xì)胞總數(shù)、嗜酸性粒細(xì)胞百分比、氣道周?chē)装Y細(xì)胞浸潤(rùn)評(píng)分、氣道壁厚度、氣道黏液物質(zhì)陽(yáng)性相對(duì)著色面積、BALF細(xì)胞因子TGF-β1水平、肺組織TGF-β1蛋白及TGF-β1 m RNA表達(dá)水平均較NS組增高(P0.01),同AS組相比均明顯降低(P0.01);AS+SCMC組氣道周?chē)z原纖維沉積面積較AS組有明顯降低(P0.01),AS+DEX組與AS組相比較無(wú)明顯差異(P0.05);AS+SCMC組與AS+DEX組相比氣道黏液物質(zhì)陽(yáng)性相對(duì)著色面積、BALF細(xì)胞因子TGF-β1表達(dá)水平無(wú)明顯差異(P0.05),氣道周?chē)装Y細(xì)胞評(píng)分差異有統(tǒng)計(jì)學(xué)意義(P0.05),其余指標(biāo)均有顯著統(tǒng)計(jì)學(xué)差異(P0.01)。相關(guān)性分析顯示在羧甲司坦組中氣道周?chē)z原纖維沉積面積與TGF-β1 m RNA表達(dá)呈正相關(guān)(r=0.82,P0.01)。結(jié)論:1.以O(shè)VA腹腔聯(lián)合皮下注射致敏和霧化吸入激發(fā)可成功復(fù)制小鼠哮喘模型。2.羧甲司坦可減輕哮喘小鼠氣道重塑,其機(jī)制可能是通過(guò)抑制TGF-β1的表達(dá)實(shí)現(xiàn)的。
[Abstract]:Objective: by ovalbumin (OVA) sensitized asthmatic mice model, to observe the effect of intervention on airway remodeling in asthmatic mice carbocysteine transforming growth factor beta 1 mediated. Methods: 32 BALB/c female mice were raised under normal circumstances after 1 weeks, mice were randomly divided into normal control (group NS), asthma model group (AS), carbocysteine group (AS+SCMC), dexamethasone group (AS+DEX) 4 groups, 8 rats in each group. The asthma group, carbocysteine intervention group, dexamethasone group on day 1,13 respectively by OVA intraperitoneal combined with subcutaneous injection of sensitized 19d 10%OVA solution to continuous inhalation of 18D 24h after the last challenge, were each 30min before stimulation group AS+SCMC received carbocisteine (10mg/kg) gavage treatment, AS+DEX group were given intraperitoneal injection of dexamethasone (2mg/kg), AS group with normal saline group.NS and treatment of atomization by normal saline. Mice were measured by BA The total number of LF cells and cell count, pathological changes of lung tissues were observed with HE staining, AB-PAS staining of airway epithelial goblet cells and mucus substance changes observed around the airway collagen deposition of MASSON staining, ELISA level detection of BALF cell factor TGF- beta 1, beta 1 were detected in the lung tissue of TGF- protein and M Expression of RNA immune group staining and real-time fluorescence quantitative PCR. Results: AS+SCMC group, AS+DEX group, BALF cell count, the percentage of eosinophils, airway inflammatory cell infiltration score, airway wall thickness, airway mucus material positive opposite color area, BALF cell factor TGF- beta 1, beta 1 lung tissue TGF- protein and TGF- beta 1 m the expression level of RNA were higher than those in group NS (P0.01), compared with the AS group were significantly decreased (P0.01); AS+SCMC group around the airway collagen deposition area compared with the AS group significantly decreased (P0.01), AS+ DEX group and AS group compared with no tomorrow Significant difference (P0.05); AS+SCMC group and AS+DEX group compared to the airway mucus material positive relative tinting area, BALF cell factor TGF- beta 1 expression level had no significant difference (P0.05), there was statistical significance score of airway inflammation (P0.05), the remaining differences around the indexes were significantly statistical difference (P0.01). Correlation analysis showed that in carbocysteine group around the airway collagen deposition area and TGF- beta 1 m RNA expression was positively correlated (r=0.82, P0.01). Conclusion: 1. OVA intraperitoneal combined with subcutaneous injection of sensitized and aerosol inhalation can successfully establish the mouse asthma model.2. carbocysteine can reduce the airway remodeling in asthmatic mice, its mechanism may be achieved by inhibiting the expression of TGF- beta 1.

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R562.25

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