【作者】 晏斌林；

【导师】 吴肖叶；

【作者基本信息】 江西医学院 ， 内科学， 2005， 硕士

【摘要】 [目的]:研究哮喘豚鼠气道上皮细胞阴离子交换蛋白2(anion exchanger 2, AE2)的表达及其意义。[方法]:(1)过敏性支气管哮喘模型的制作:取20 只豚鼠随机分为2 组,卵清白蛋白(OVA)后腿肌肉注射与雾化吸入诱发豚鼠哮喘。在连续激发3 天后,测定激发前后呼吸频率;(2)测定哮喘豚鼠支气管粘膜周围肺组织嗜酸性粒细胞(EOS)浸润情况;支气管肺泡灌洗液(BALF)中EOS 数量;血IgE 浓度测定和EOS 计数;(3)取豚鼠气道上皮,以免疫印迹(western blotting)和逆转录-酶联扩增反应(RT-PCR)法作AE2蛋白及其mRNA 表达水平的检测。[结果]:①呼吸频率测定:实验第14 天对照(Control)组和哮喘(Asthma)组豚鼠呼吸均较平稳,经激发后两组则有明显区别。激发前Control 组(105.4±7.63次/分)与Asthma组(108.8±6.92次/分)比较无显著性差异(p>0.05);激发后Control 组(106.8±5.67 次/分)与Asthma 组(130.5±7.86 次/分)比较有显著性差异(p<0.01);同组内比较Control 组无显著性差异(p>0.05),Asthma 组比较有显著性差异(p<0.01)。②血清IgE 浓度和血EOS 计数测定:Control 组IgE 浓度(72.7±27.79)与Asthma 组的IgE(390.6±154.16)比较,有显著性差异(p<0.01)。Control 组EOS 计数(0.051±0.017)与Asthma 组(0.265±0.024)比较,有显著性差异(p<0.01)。③豚鼠肺组织病理形态改变:光学显微镜观察发现:Control 组,豚鼠肺组织切片支气管粘膜上皮细胞排列整齐.管腔内未见上皮细胞脱落及炎性渗出物,管腔及肺泡间隔可见少许淋巴细胞浸润,未见嗜酸性粒细胞.Asthma 组,豚鼠肺组织切片支气更多还原

【Abstract】 [Objective]: To study expression and its implication of AE2 protein and mRNA in epithelium of asthmatic guinea pigs. [Method]: To use 20 guinea pigs and divide them into two groups at random without distinction of sex. One group names Control group( n=10) and the other Asthma one(n=10). The antigen-sensitized and -challenged guinea pig model of allergic asthma is made after consecutive 3d challenge. To detect pre-and post-challenge respiratory freqency until 3d postchallenge; To observe eosinophil(EOS) infiltration around bronchial mucous membrane and pulmonary tissue in asthmatic model; The number of eosinophils in bronchoalveolar lavage fluid is also observed; To detect the concentratin of serum IgE and count eosinophil of blood. Using the epithelium in airway of guinea pig, to examine the levels of AE2 protein and mRNA by means of western blotting and RT-PCR technique. [Results]: ①the detection of respiratory frequency: The ovalbumin (OVA)-sensitized guinea pigs and the Control group both have a steady breath on the 14th day in experiment , there are obvious differences after exciting. The Control group(105.4±7.63bpm ) do not relatively have a significant difference (p> 0.05 ) with the Asthma group (108.8±6.92bpm) before exciting; The Control group (106.8±5.67bpm ) relatively has a significant difference (p<0.01 ) with the Asthma group (130.5 ±7.86bpm ) after challenge; Not having significant difference (p> 0.05 ) to compare in the same Control group, the Asthma group relatively has a significant difference (p<0.01 ).②the detection of serum IgE and count eosinophil of blood: Compared with the concentration of IgE of the Control group (72.7±27.79 ) that (390.6±154.16 ) of the Asthma group has significant difference (p<0.01); There is significant difference (p <0.01) between the Control group( 0.051±0.017)and the Asthma group (0.265±0.024 )in EOS count.③pulmonary tissue pathologically morphological change: We find, under the optics microscope,that the Control group, guinea pig lung tissue slices bronchus membranes’epithelial cells arrange neatly. There are no epithelial cells to come off and no inflammation cells out of the ducts, a little lymphocytes are seen in ducts ,not finding eosinophil infiltration .But the asthma group has a distinct picture: the epithelial cells of bronchus membranes arrange disorderlily . Some epithelium come off into the airway ducts .In alveolus interval we can see more lymphocytes and eosinphilic granulocytes soakage .The basolateral membranes become thicker. Bronchus duct is hard to get at all even to dwindle, the mucus blocks the airway ducts partly ④the levels of AE2 protein and mRNA:western blotting reveals that AE2 protein has a basic expression in the Control group. AE2 protein expression of the Asthma group(168287.0 ±28663.68) decreases obviously compared with the Control group(244350.4±29358.02)(P<0.01). This is true of AE2 mRNA expression. Compared with the Control group(0.852±0.0107), the level of AE2 mRNA of the Asthma group decreases obviously(0.790±0.0104)(P<0.01). [Conclusions]: 1、Guinea pig’s allergic bronchus asthma models appear obvious dyspnea, wheeze; EOS and serum IgE obviously rise in the blood and BALF;Obvious pathologically morphological change appears. 2、AE2 protein and mRNA have their basic expression in normal guinea pig.In guinea pig asthma models,expressions of their epithelium AE2 protein and mRNA downregulate.更多还原