间充质干细胞对哮喘大鼠气道重塑的治疗作用及机制研究

发布时间：2018-03-28 08:55

本文选题：支气管哮喘　切入点：气道重塑　出处：《山西医科大学》2017年硕士论文

【摘要】：目的:观察间充质干细胞(MSCs)对转化生长因子-β1(TGF-β1)致上皮细胞-间质转化作用的影响,探讨MSCs对哮喘气道重塑的治疗作用及可能的作用机制。方法:40只Wistar雄性大鼠分为四组:A.对照组、B.模型组、C.MSCs组、D.布地奈德组。除A组外,其余各组分别在第1、第8天采用10%卵白蛋白抗原混悬液1ml进行腹腔注射致敏大鼠,第15天开始超声雾化1%卵白蛋白激发,20min/d,共8周;C组于第35、第45及第55天激发前30min尾静脉注射脐带MSCs 0.2ml(1×106个/ml);D组于每次激发前2h雾化吸入布地奈德2mg;A组以0.9%氯化钠溶液代替卵白蛋白,其余干预方法同B组。在最后一次激发后的二十四小时之内,收集血清、支气管肺泡灌洗液和肺组织,HE染色观察气道重塑病理改变,支气管壁和平滑肌厚度由图像分析软件计算得出;支气管肺泡灌洗液及血清中TGF-β1含量由酶联免疫吸附试验检测;肺组织E-钙黏蛋白、α-平滑肌肌动蛋白、纤维连接蛋白的表达由SABC法检测。结果:B组大鼠支气管壁及平滑肌厚度[(42.36±1.50、21.01±0.10)μm]显著厚于A组[(25.69±0.27、14.97±0.13)μm];支气管肺泡灌洗液及血清中TGF-β1含量[(21.01±0.18、70.97±0.23)μg/L]显著高于A组[(1.14±0.19、23.05±0.16)μg/L];上皮标志物E-钙黏蛋白(0.256±0.087)表达显著低于A组(0.469±0.012),间质标志物α-平滑肌肌动蛋白、纤维连接蛋白[(0.521±0.030、0.527±0.022)]表达显著高于A组[(0.371±0.007、0.387±0.020)](均P0.01)。C、D组大鼠支气管壁及平滑肌厚度分别为32.40±2.50μm、35.86±0.55μm和15.71±0.89μm、18.72±0.89μm,支气管肺泡灌洗液及血清中TGF-β1含量分别为3.53±0.43 ug/L、3.25±0.25 ug/L和31.07±0.89 ug/L、32.08±0.18 ug/L,均显著低于B组(均P0.01);E-钙黏蛋白表达[(0.308±0.023)、(0.296±0.010)]显著高于B组,α-平滑肌肌动蛋白和纤维连接蛋白表达[(0.438±0.057)、(0.445±0.027)和(0.459±0.041)、(0.458±0.029)]显著低于B组(均P0.01)。结论:1、TGF-β1诱导的上皮细胞-间质转化是哮喘气道重塑的机制之一;2、MSCs通过抑制TGF-β1诱导的上皮细胞-间质转化,对支气管哮喘气道重塑发挥治疗作用。
[Abstract]:Aim: to observe the effect of mesenchymal stem cells (MSCs) on epithelial cell-mesenchymal transformation induced by transforming growth factor- 尾 _ 1- TGF- 尾 _ 1 (TGF- 尾 _ 1). To investigate the therapeutic effect and possible mechanism of MSCs on airway remodeling in asthmatic rats, 40 Wistar male rats were divided into four groups: control group, control group, C.MSCs group, D 路budesonide group, with the exception of group A. The other groups were sensitized by intraperitoneal injection of 10% ovalbumin antigen suspension (1ml) on the 1st and 8th day, respectively. On the 15th day, 1% ovalbumin was stimulated by ultrasound for 20 min / d. After 8 weeks, 30min caudal vein injection of MSCs 0.2ml(1 脳 106 / ml / d was performed in group C at 35th, 45th and 55th day. Group D inhaled budesonide 2 mg / g of budesonide and inhaled 2 mg of budesonide with 0.9% sodium chloride 2 h before stimulation. Instead of ovalbumin, The other intervention methods were the same as group B. within 24 hours after the last stimulation, the serum, bronchoalveolar lavage fluid and lung tissue were collected to observe the pathological changes of airway remodeling. The thickness of bronchial wall and smooth muscle was calculated by image analysis software; the content of TGF- 尾 1 in bronchoalveolar lavage fluid and serum was measured by enzyme linked immunosorbent assay (Elisa); Results the thickness of bronchial wall and smooth muscle in group B was significantly thicker than that in group A [25.69 卤0.270.97 卤0.13 渭 m], and the content of TGF- 尾 _ 1 in bronchoalveolar lavage fluid and serum [21.01 卤0.1870.97 卤0.23 渭 g / L] was significantly higher than that in group A [1.14 卤0.19 卤23.05 卤0.16 渭 g / L]. The expression of 伪 -smooth muscle actin was significantly lower in group A than in group A (0.256 卤0.087), and the expression of 伪 -smooth muscle actin was significantly lower than that in group A (0.469 卤0.012). The expression of fibronectin [0.521 卤0.030 卤0.527 卤0.022] was significantly higher than that in group A [0.371 卤0.007 卤0.387 卤0.020] (the thickness of bronchial wall and smooth muscle in group A was 32.40 卤2.50 渭 m, 35.86 卤0.55 渭 m and 15.71 卤0.89 渭 m, 18.72 卤0.89 渭 m, respectively). The levels of TGF- 尾 _ 1 in bronchoalveolar lavage fluid and serum were 3.53 卤0.43 渭 L ~ 3.25 卤0.25 ug/L and 31.07 卤0.89 卤0.89 渭 m, respectively, which were significantly lower than those in group B (P < 0.05). The expressions of 伪 -smooth muscle actin and fibronectin [0.438 卤0.0577.45 卤0.027] and 0.459 卤0.041 (0.458 卤0.029) were significantly lower in group B than those in group B (all P 0.01). Conclusion the expression of 伪 -smooth muscle actin and fibronectin is the mechanism of airway remodeling in asthma. Mesenchymal mesenchymal transformation induced by TGF- 尾 1 was inhibited by TGF- 尾 1. To play a therapeutic role in bronchial asthma airway remodeling.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R562.25

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