TGF-β对肥胖哮喘小鼠气道重塑的影响及吡非尼酮的干预作用

发布时间：2018-05-15 09:02

本文选题：哮喘 + 肥胖　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:探讨TGF-β对肥胖哮喘小鼠气道重塑的影响及吡非尼酮干预的作用。方法:将75只健康雌性C57BL/6J小鼠随机均分为:对照组(A组)、肥胖组(B组)、肥胖哮喘组(C组)、布地奈德治疗组(D组)、吡非尼酮治疗组(E组)。肥胖组小鼠均采用高脂饮食诱导肥胖,其中C、D、E组小鼠采用卵白蛋白腹腔注射致敏联合雾化吸入激发的方法构建哮喘模型,对照组小鼠均采用普通饲料饲养,同期给予生理盐水致敏激发。布地奈德治疗组小鼠给予布地奈德悬液雾化吸入(0.5mg/ml,30min/次,qd),吡非尼酮治疗组给以吡非尼酮(300mg/kg)饮水干预,其余组给予正常饮水。治疗4周后,收集小鼠支气管肺泡灌洗液(BALF)进行细胞计数及分类。采用HE、PAS、Masson染色观察肺组织病理切片的气道炎症、气道重塑及气道粘液分泌情况。采用ELISA法测定血清中转化生长因子β(TGF-β)浓度,取右肺组织用Westernblot检测TGF-β的表达变化情况。应用计算机图像分析软件技术测定气管壁总面积(WAt)、气道平滑肌面积(WAm)和管腔基底膜周长(Pbm)。结果:试验过程中,肥胖组小鼠体质量明显高于空白对照组(P0.05)。致敏激发过程中,哮喘各组小鼠均有不同程度的哮喘急性发作表现。至小鼠干预结束时,各组小鼠均无死亡。肥胖哮喘组小鼠BALF中白细胞总数、中性粒细胞及嗜酸性细胞比例均明显高于其余四组;肥胖哮喘小鼠血清、肺组织中TGF-β的水平、气管壁厚度(WAt/Pbm)、平滑肌厚度(WAm/Pbm)均高于对照组、肥胖组、布地奈德治疗组、吡非尼酮干预组。肥胖组BALF白细胞总数、嗜酸性粒细胞比例均略高于对照组,差异无统计学意义(P0.05);吡非尼酮干预组及布地奈德治疗组小鼠BALF的细胞总数、中性粒细胞比例和嗜酸性粒细胞比例较肥胖哮喘组均明显减少(P0.05),吡非尼酮干预组BALF中嗜酸性粒细胞比例较布地奈德治疗组减少(P0.05),但两组中性粒细胞比例下降不明显(P0.05);吡非尼酮干预组及布地奈德治疗组WAt/Pbm、WAm/Pbm均较肥胖哮喘组明显降低(P0.05),吡非尼酮干预组WAt/Pbm较布地奈德治疗组降低(P0.05),WAm/Pbm两组比较差异无统计学意义(P0.05);小鼠血清中及肺组织中TGF-β水平按照肥胖哮喘组、布地奈德治疗组、吡非尼酮治疗组、肥胖组、对照组依次递减(P0.05),吡非尼酮干预组下降最为明显(P0.05)。结论:TGF-β在肥胖哮喘小鼠气道中高表达,其高表达与气道炎症、气道高分泌及气道重塑有密切关系,吡非尼酮能有效抑制气道炎症,达到改善气道炎症及气道重塑的作用。
[Abstract]:Aim: to investigate the effect of TGF- 尾 on airway remodeling in obese asthmatic mice and the effect of pifenidone. Methods: 75 healthy female C57BL/6J mice were randomly divided into three groups: control group A, obesity group B, obese asthma group C, budesonide treatment group D and pifenidone treatment group. Obesity was induced by high-fat diet in all the obese mice. The asthmatic model was established by intraperitoneal injection of ovalbumin and aerosol inhalation in mice in CfD E group, and the normal diet was used in all the mice in the control group. At the same time, saline was given to sensitize and excite. The mice in budesonide group were given budesonide suspension aerosol inhalation (0.5 mg / ml) for 30 min / time, pifenidone group was treated with pifenidone (300 mg / kg) drinking water, and the rest group was given normal drinking water. After 4 weeks of treatment, BALF was collected from mouse bronchoalveolar lavage fluid for cell count and classification. Airway inflammation, airway remodeling and airway mucus secretion were observed by PAS-Masson staining. The concentration of TGF- 尾 in serum was measured by ELISA method, and the expression of TGF- 尾 in right lung tissue was detected by Westernblot. The total area of trachea wall, airway smooth muscle area (Wam) and the circumference of basal membrane of the tube were measured by computer image analysis software. Results: the body weight of obese group was significantly higher than that of control group (P 0.05). In the course of sensitizing and arousing, the asthmatic mice in each group showed different degrees of acute asthma attack. At the end of the intervention, there was no death in all groups. The percentage of leukocytes, neutrophils and eosinophils in BALF of obese asthmatic mice was significantly higher than that of the other four groups, and the levels of TGF- 尾 in serum and lung tissue, thickness of trachea wall and thickness of smooth muscle were significantly higher in obese asthmatic mice than in control group. Obesity group, budesonide treatment group, pifenidone intervention group. The total number of BALF leukocytes and the percentage of eosinophil in obese group were slightly higher than those in control group, the difference was not statistically significant (P 0.05), the total number of BALF cells in pifenidone intervention group and budesonide treatment group was higher than that in control group. The percentage of neutrophils and eosinophil in BALF in the pifenidone intervention group was lower than that in the budesonide treatment group, but the percentage of neutrophils in both groups was not clear, but the percentage of eosinophilic granulocytes in the treatment group was significantly lower than that in the obese asthmatic group. Compared with obese asthma group, the WAt/Pbm of pifenidone intervention group and budesonide treatment group was significantly lower than that of obese asthmatic group, and the WAt/Pbm of pifenidone intervention group was lower than that of budesonide treatment group. There was no significant difference between the two groups. The levels of TGF- 尾 in the tissue were determined according to the obesity asthma group. Budesonide treatment group, pifenidone treatment group, obesity group, the control group decreased in turn (P 0.05), pifenidone intervention group decreased the most significantly (P 0.05). Conclusion the high expression of TGF- 尾 in the airway of obese asthmatic mice is closely related to airway inflammation, airway hypersecretion and airway remodeling. Pifenidone can effectively inhibit airway inflammation and improve airway remodeling.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R562.25

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