羧甲司坦对哮喘小鼠转化生长因子-β1介的气道重构的影响

发布时间：2018-03-09 19:17

本文选题：哮喘　切入点：小鼠　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过鸡卵清白蛋白(OVA)致敏的哮喘小鼠模型,观察羧甲司坦干预对哮喘小鼠转化生长因子-β1介导的气道重构的影响。方法:将32只BALB/c雌性小鼠在正常环境下饲养1周后,采用随机法将小鼠分为正常对照组(NS)、哮喘模型组(AS)、羧甲司坦组(AS+SCMC)、地塞米松组(AS+DEX)4组,每组8只。哮喘组、羧甲司坦干预组、地塞米松干预组于第1、13天分别以OVA腹腔联合皮下注射致敏,第19d开始予10%OVA溶液连续雾化激发18d,末次激发24h后取材,每次激发前30min AS+SCMC组予羧甲司坦(10mg/kg)灌胃处理,AS+DEX组予以地塞米松(2mg/kg)腹腔注射,AS组以生理盐水代替。NS组雾化及处理全部以生理盐水代替。测定各组小鼠BALF细胞总数和细胞分类计数,HE染色观察肺组织病理学改变,AB-PAS染色观察气道上皮杯状细胞和黏液物质改变,MASSON染色观察气道周围胶原纤维沉积,ELISA检测BALF细胞因子TGF-β1的水平,免疫组化染色和实时荧光定量PCR分别检测肺组织TGF-β1蛋白和m RNA表达水平。结果:AS+SCMC组、AS+DEX组BALF细胞总数、嗜酸性粒细胞百分比、气道周围炎症细胞浸润评分、气道壁厚度、气道黏液物质阳性相对着色面积、BALF细胞因子TGF-β1水平、肺组织TGF-β1蛋白及TGF-β1 m RNA表达水平均较NS组增高(P0.01),同AS组相比均明显降低(P0.01);AS+SCMC组气道周围胶原纤维沉积面积较AS组有明显降低(P0.01),AS+DEX组与AS组相比较无明显差异(P0.05);AS+SCMC组与AS+DEX组相比气道黏液物质阳性相对着色面积、BALF细胞因子TGF-β1表达水平无明显差异(P0.05),气道周围炎症细胞评分差异有统计学意义(P0.05),其余指标均有显著统计学差异(P0.01)。相关性分析显示在羧甲司坦组中气道周围胶原纤维沉积面积与TGF-β1 m RNA表达呈正相关(r=0.82,P0.01)。结论:1.以OVA腹腔联合皮下注射致敏和雾化吸入激发可成功复制小鼠哮喘模型。2.羧甲司坦可减轻哮喘小鼠气道重塑,其机制可能是通过抑制TGF-β1的表达实现的。
[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.

【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R562.25

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