HIF-1α在变应性鼻炎鼻黏膜炎症及组织重塑中的作用研究

发布时间：2018-04-02 18:15

本文选题：变应性鼻炎　切入点：鼻黏膜　出处：《南京医科大学》2013年博士论文

【摘要】：第一部分变应性鼻炎小鼠急性和慢性模型的建立及评价目的建立符合本课题要求的变应性鼻炎(AR)小鼠急性和慢性模型。方法采用卵清蛋白(OVA)全身致敏BALB/c、鼠后,8天OVA激发剂连续滴鼻激发出AR急性模型,随之持续14周的间断激发诱导出AR慢性模型,并从鼻部症状学、鼻黏膜组织病理学(HE染色、AB-PAS染色和Masson染色)及血清特异性IgE水平(ELISA法)这三个方面来评价AR动物模型建立的成功与否。结果AR小鼠急性和慢性模型都显示出了OVA致敏导致的炎症反应表现,包括鼻部症状、炎性细胞鼻黏膜浸润、黏膜下嗜酸性粒细胞(EOS)募集及血清中OVA特异性IgE水平上升等。慢性AR组动物鼻黏膜的杯状细胞数目及胶原纤维沉积面积均明显高于急性AR组及对照组。结论本实验制备出的AR急慢性动物模型均符合AR的病理生理特征,而且AR慢性模型具备了鼻黏膜组织重塑的主要特征。第二部分调控HIF-1α对变应性鼻炎小鼠急性模型鼻黏膜炎症的影响目的探讨低氧诱导因子1α (HIF-1α)在AR小鼠急性模型鼻黏膜炎症中的作用。方法通过OVA全身致敏和局部激发BALB/c小鼠,制备AR小鼠急性模型,并对其应用HIF-1α抑制剂2-甲氧基雌二醇(2ME2)和缺氧模拟剂氯化钻(CoCl2)进行预处理,检测小鼠鼻黏膜中HIF-1α和血管内皮生长因子(VEGF)的含量,同时评估多项反映鼻部变应性炎症反应的指标。结果研究发现在AR小鼠模型的鼻黏膜中HIF-1α和VEGF水平显著升高,AR小鼠急性模型都显示出了OVA致敏导致的炎症反应表现,包括鼻部症状、炎性细胞鼻黏膜浸润及EOS募集、鼻腔灌洗液中的IL-4和IL-5以及血清中OVA特异性IgE水平上升等。2ME2可以抑制鼻黏膜中HIF-1α和VEGF的表达和上述OVA致敏导致的炎症反应,而缺氧模拟剂CoCl2可以增强这些反应。HIF-1α在鼻部的表达水平与AR小鼠鼻部炎症的严重程度相关。结论HIF-1α的活化参与了AR的发病机制,抑制HIF-1α可能成为AR治疗的一个新方向。第三部分调控HIF-1α对变应性鼻炎小鼠慢性模型鼻黏膜炎症及组织重塑的影响目的探讨HIF-1α及其调控的生长因子(VEGF、FGF-2、TGF-β1)在AR小鼠慢性模型的鼻黏膜炎症和组织重塑中的作用。方法通过OVA全身致敏并长期局部激发BALB/c小鼠,制备AR小鼠慢性模型,并对其应用2ME2和CoCl2进行预处理,检测小鼠鼻黏膜中HIF-1α, VEGF、FGF-2和TGF-β1的含量,同时评估多项反映鼻部变应性炎症反应的指标以及鼻黏膜重塑的组织病理学表现。结果AR小鼠慢性模型的鼻黏膜中HIF-1α蛋白水平显著升高,除了OVA激发导致的鼻部变应性炎症反应表现以外,病理切片显示出鼻黏膜组织重塑的重要特征,包括杯状细胞大量化生、胶原纤维沉积明显增强和上皮细胞大量脱落等。2ME2可以抑制鼻黏膜中HEF-1α的表达,减轻鼻黏膜炎症反应和组织重塑程度；而长期应用CoCl2对AR小鼠慢性模型的干预并不能获得预期的明显效果。AR小鼠急慢性模型鼻黏膜中的HIF-1α蛋白水平与VEGF、FGF-2和TGF-β1的蛋白水平明显相关。结论AR小鼠慢性模型中HIF-1α水平与鼻黏膜炎症和组织重塑程度密切相关。长期2ME2干预可通过对HIF-1α的抑制而有效改善持续OVA刺激所致的鼻黏膜组织重塑,其机制可能与下调VEGF、FGF-2和TGF-β1的水平有关。
[Abstract]:The first part: establishment and evaluation of acute and chronic allergic rhinitis mice model objective: to establish the requirements of the subject of allergic rhinitis (AR) mouse model of acute and chronic. Methods using ovalbumin (OVA) sensitized BALB/c rats body, after 8 days of continuous nasal OVA activator excited by acute AR model, will last for 14 weeks the intermittent excitation induced AR in chronic model, and from the nasal symptoms, nasal mucosa histopathology (HE staining, AB-PAS staining and Masson staining) and serum specific IgE level (ELISA) of these three aspects to evaluate the animal model of AR is built and the success of AR mice. The results of acute and chronic model show the inflammatory reaction in OVA sensitized to the performance, including nasal symptoms, inflammatory cell infiltration in nasal mucosa, submucosal eosinophils (EOS) and raised serum OVA specific IgE levels increased. AR group of nasal mucosa of chronic animal goblet The number of cells and collagen deposition area were significantly higher than those in acute AR group and control group. Conclusion the experiment for the preparation of acute and chronic AR animal model were consistent with the pathophysiological features of AR, and the AR model has the main features of chronic nasal mucosa remodeling. The second part is the regulation of HIF-1 alpha in acute mouse model of allergic rhinitis objective to investigate the nasal mucosal inflammation of hypoxia inducible factor 1 alpha (HIF-1 alpha) in acute AR mice nasal mucosa inflammation. Methods by OVA systemic sensitization and local stimulation of BALB/c mice, preparation of acute AR mice model, and the use of HIF-1 alpha inhibitor 2- methoxyestradiol (2ME2) and hypoxia mimetic cobalt chloride (CoCl2) pretreatment, detection of HIF-1 in the nasal mucosa of the mice alpha and vascular endothelial growth factor (VEGF) were assessed at the same time, a number reflecting the nasal allergic inflammation index. The research results found In the nasal mucosa of AR mice model of HIF-1 alpha and VEGF levels were significantly elevated in acute AR mice showed inflammation caused by OVA sensitization, including nasal symptoms, nasal mucosa inflammatory cell infiltration and EOS to raise the inflammation in the nasal lavage fluid of IL-4 and IL-5 and serum OVA specific IgE the level of rising.2ME2 can inhibit the expression of HIF-1 and VEGF in nasal mucosa and the OVA sensitized to the severity of hypoxia simulating agent CoCl2 can enhance the reaction of.HIF-1 alpha in the nose and expression of AR in mice nasal inflammation. Conclusion the HIF-1 alpha activation involved in the pathogenesis of AR, inhibition of HIF-1 alpha may become a new target for the treatment of AR. Objective to investigate the growth factor HIF-1 alpha and control effect of third part of regulation of HIF-1 alpha on the model of chronic nasal mucosa inflammation and tissue remodeling in mice with allergic rhinitis (VEGF, FGF-2, T GF- beta 1) in nasal mucosa of chronic inflammation and tissue remodeling in AR mice model in vivo. Methods OVA systemic sensitization and long-term local stimulation in BALB/c mice, AR mice were prepared for chronic model, and the application of 2ME2 and CoCl2 pretreatment, detection of HIF-1 in the nasal mucosa of the mice alpha, VEGF, and FGF-2 content TGF- beta 1, while evaluating a number reflecting the performance of nasal allergic inflammation of the nasal mucosa remodeling index and pathology. Results the level of HIF-1 protein in nasal mucosa of chronic AR mouse model increased significantly, in addition to the nasal allergic inflammatory reaction caused by the excitation of OVA, pathological section showed important features of nasal mucosa tissue remodeling, including a large number of goblet cells, the expression of collagen deposition was significantly enhanced and a large number of epithelial cells shedding.2ME2 can inhibit HEF-1 alpha in nasal mucosa, reduce nasal inflammation and tissue The degree of plastic; while the long-term application of CoCl2 on AR mice model of chronic intervention can obtain the HIF-1 protein levels and VEGF expected significant effect of.AR mice model of acute and chronic nasal mucosa, significantly related protein levels of FGF-2 and TGF- beta 1. Conclusion AR is closely related to the model of mice with chronic HIF-1 alpha level and nasal mucosal inflammation and the degree of tissue remodeling. The long-term intervention of 2ME2 by inhibiting HIF-1 alpha and effectively improve the sustained stimulation of OVA nasal mucosa remodeling caused by, and to investigate the possible mechanism of VEGF, the levels of FGF-2 and TGF- beta 1.

【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R765.21

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