瞬时受体电位通道和神经肽表达上调在甲醛诱导型支气管哮喘小鼠模型中的作用

发布时间：2018-05-24 10:54

本文选题：甲醛 + 过敏性哮喘　；参考：《华中师范大学》2013年博士论文

【摘要】：甲醛是一种常见的装修型化学性室内空气污染物,也是一种全球性的环境污染物。它具有污染来源广、时间长、水平高、毒性种类多等特点。甲醛对健康的影响主要包括：呼吸道和眼部急性刺激作用、神经行为改变、生殖毒性、免疫毒性、氧化损伤、遗传毒性等诸多方面。近年来,世界各地哮喘发病率都在快速增长,但致敏原的种类和数量并未发生显著变化。非过敏原性空气污染物的大量增加被认为是原因之一,但其分子机制尚不清楚。不断有研究表明,甲醛暴露可以引起哮喘或哮喘样症状。为了探讨甲醛是否可以诱发哮喘和其可能的发病机理,本论文进行了如下体内研究： 1.甲醛导致中枢神经系统神经源性炎症及氧化性损伤发生：对Balb/c小鼠连续甲醛气态暴露7天,每天8小时,设(a)对照组；(b)0.5mg/m3甲醛组；(c)3.0mg/m3甲醛组,在3.0mg/m3水平甲醛作用下,与对照组相比较：(1)脑组织谷胱甘肽(GSH)含量呈极显著性降低(p0.01),脊髓组织GSH含量呈显著性降低(p0.05)；(2)脑组织丙二醛(MDA)含量呈极显著性增高(p0.01),脊髓组织MDA含量呈显著性增高(p0.05)；(3)脑组织促炎症细胞因子白介素-IL-1β含量呈显著性增高(p0.05),脊髓组织IL-1β含量呈极显著性增高(p0.01)；(4)脑组织速激肽物质P含量(substance P)呈极显著增高(p0.01),脊髓组织substance P含量呈极显著性增高(p0.01)。本实验说明甲醛可以导致中枢神经系统神经性炎症及氧化损伤发生。 2. TRPA1介导甲醛所致气道炎症发生：对Balb/c小鼠连续甲醛气态暴露10天,每天8小时。设(a)对照组；(b)0.5mg/m3甲醛组；(c)1.0mg/m3甲醛组；(d)3.0mg,/m3甲醛组；(e)3.0mg/m3+HC-030031[瞬时受体电位通道亚型A1(TRPA1)拮抗剂]组,发现在3.0mg/m3甲醛水平作用下,与对照组相比较：(1)血清中免疫球蛋白E(1gE)水平显著上升(p0.01)；(2)小鼠肺泡灌洗液中白介素-4(IL-4)含量显著上调(p0.01)；(3)血清中超氧化物歧化酶(SOD)活力显著下降(p0.01),MDA含量显著上升(p0.01)；对3.0mg/m3甲醛给予HC-030031(50mg/kg)处理,发现IgE水平下调(p0.01),IL-4水平下调(p0.01), SOD活力上升(p0.05),MDA含量下降(p0.05)；本实验说明TRPA1可以介导甲醛所致气道炎症发生和氧化损伤作用。 3. TRPA1和TRPV1通道蛋白介导气态甲醛诱导哮喘作用：对Balb/c雄性小鼠甲醛动态式气态暴露4周,每天6小时,卵清蛋白(OVA)致敏并激发1周建立甲醛诱导哮喘模型。设(a)对照组；(b) OVA组；(c)3.0mg/m3甲醛组；(d)3.Omg/m3甲醛+OVA组；(e)3.0mg/m3甲醛+OVA+HC-030031组；(f)3.0mg/m3甲醛+OVA+capsazepine(瞬时受体电位通道亚型V1拮抗剂)组,分别测定气道高反应性、气道重塑、气道炎症相关指标。与对照组相比,甲醛组气道反应性和肺部病理学变化并不明显,但嗜酸性粒细胞计数显著增高(p0.05),血清总IgE含量明显上调(p0.01),IL-4含量显著升高(p0.01), substance P表达上调(p0.01),降钙素基因相关肽(CGRP)表达上调(p0.05),IL-1p水平升高(p0.01)；与OVA组相比,甲醛+OVA组呈现显著气道高反应性和气道重塑现象,嗜酸性粒细胞计数显著增高(p0.01),血清中总IgE水平显著上调(p0.01),肺组织IL-4含量显著升高(p0.01)；substance P表达上调(p0.01)、CGRP表达上调(p0.01), IL-1β水平升高(p0.01)。对FA+OVA组给予TRPA1通道拮抗剂HC-030031和TRPV1通道拮抗剂CPZ,结果显示：拮抗组与模型组相比,(1)气道高反应性显著降低：(2)肺组织病理变化明显好转；(3)小鼠肺泡灌洗液中嗜酸性粒细胞计数降低(p0.01),肺组织IL-4含量显著降低(p0.01),血清中IgE水平降低(p0.01)；(4)substance P表达下调(p0.01)、CGRP表达下调(p0.05), IL-1β水平无显著变化(p0.05)。结果说明甲醛可以提高哮喘的发病风险,它可以直接导致神经源性气道炎症,并以免疫佐剂身份加重OVA过敏原导致气道炎症；Substance P和CGRP参与甲醛诱导哮喘的病理生理过程,TRPA1和TRPV1通道蛋白介导气态甲醛诱导哮喘作用并调控神经肽的释放。
[Abstract]:Formaldehyde is a common decoration chemical indoor air pollutant and a global environmental pollutant. It has a wide range of pollution sources, long time, high level, and many toxic types. The effect of formaldehyde on health mainly includes: respiratory and ocular acute stimulation, neurobehavioral changes, reproductive toxicity, immunotoxicity, oxygen, and oxygen. In recent years, the incidence of asthma in all parts of the world has been growing rapidly, but the variety and number of allergens have not changed significantly. A large increase in non allergen air pollutants is considered to be one of the reasons, but its molecular mechanism is not clear. Asthma or asthma like symptoms. In order to investigate whether formaldehyde can induce asthma and its possible pathogenesis, the following studies were carried out in vivo.
1. formaldehyde caused neurogenic inflammation and oxidative damage in the central nervous system: 7 days of continuous formaldehyde exposure to Balb/c mice, 8 hours a day, (a) control group; (b) 0.5mg/m3 formaldehyde group; (c) 3.0mg/m3 formaldehyde group and compared with the control group under the action of 3.0mg/m3 level formaldehyde: (1) the content of glutathione (GSH) in the brain tissue was extremely significant The content of GSH in spinal cord tissue decreased significantly (P0.05), and (2) the content of malondialdehyde (MDA) in the brain tissue was significantly increased (P0.01), and the content of MDA in the spinal cord tissue was significantly increased (P0.05) (P0.05); (3) the content of interleukin -IL-1 beta in the brain tissue was significantly increased (P0.05), and the content of IL-1 beta in the spinal tissue was very significant. Increase (P0.01); (4) the content of P (substance P) of the brain tissue tachykinin (substance P) was significantly increased (P0.01), and the content of substance P in the spinal cord was significantly increased (P0.01). This experiment showed that formaldehyde could lead to neurogenic inflammation and oxidative damage in the central nervous system.
2. TRPA1 mediated formaldehyde induced airway inflammation: 10 days of continuous formaldehyde exposure to Balb/c mice, 8 hours a day. (a) control group; (b) 0.5mg/m3 formaldehyde group; (c) 1.0mg/m3 formaldehyde group; (d) 3.0mg, /m3 formaldehyde group; (E) 3.0mg/m3+HC-030031[instantaneous receptor channel subtype antagonist] group, found in formaldehyde level Under the action, compared with the control group, (1) the serum level of immunoglobulin E (1gE) increased significantly (P0.01); (2) the content of interleukin -4 (IL-4) in the mouse alveolar lavage was significantly up (P0.01); (3) the activity of superoxide dismutase (SOD) in the serum decreased significantly (P0.01) and MDA increased significantly (P0.01), and HC-030031 (P0.01) was given to 3.0mg/m3 formaldehyde (P0.01). /kg) treatment, IgE level down (P0.01), IL-4 level down (P0.01), SOD activity (P0.05) and MDA content decreased (P0.05). This experiment shows that TRPA1 can mediate the effect of formaldehyde induced airway inflammation and oxidative damage.
3. TRPA1 and TRPV1 channel proteins mediate the effect of gaseous formaldehyde induced asthma: for 4 weeks of formaldehyde dynamic exposure in Balb/c male mice, 6 hours a day, ovalbumin (OVA) sensitization and excitation of formaldehyde induced asthma model. Set (a) control group; (b) OVA group; (c) 3.0mg/m3 formaldehyde group; (d) 3.Omg/m3 formaldehyde +OVA group; (d) 3.Omg/m3 formaldehyde +OVA group; (d) 3.Omg/m3 formaldehyde +OVA group; (d) 3.Omg/m3 formaldehyde +OVA group; Group +OVA+HC-030031; (f) 3.0mg/m3 formaldehyde +OVA+capsazepine (transient receptor potential channel subtype V1 antagonist), the airway hyperresponsiveness, airway remodeling, airway inflammation related indexes respectively. Compared with the control group, the airway reactivity and pulmonary pathological changes were not obvious in the control group, but the eosinophil count increased significantly (P0.05). The content of IgE was obviously up (P0.01), the content of IL-4 increased significantly (P0.01), the expression of substance P was up (P0.01), the expression of calcitonin gene related peptide (CGRP) up up (P0.05), IL-1p level increased (P0.01). Compared with the OVA group, the formaldehyde group showed significant airway hyperresponsiveness and airway remodeling, and the eosinophil count increased significantly. The total IgE level in the serum was significantly up-regulated (P0.01), the IL-4 content in the lung tissue was significantly increased (P0.01), the expression of substance P was up (P0.01), the expression of CGRP was up (P0.01), and the IL-1 beta level increased (P0.01). The results showed that the antagonist and the channel antagonist were compared with the model group, (1) the hyperreaction of the airway. (2) the pathological changes of lung tissue were obviously improved; (3) the eosinophil count in the alveolar lavage fluid in mice decreased (P0.01), the IL-4 content in the lung tissue decreased significantly (P0.01), the level of IgE in the serum decreased (P0.01); (4) the downregulation of substance P (P0.01), CGRP expression down (P0.05), IL-1 beta level had no significant changes (P0.05). The results said It can improve the risk of asthma, which can directly cause neurogenic airway inflammation and aggravate the OVA allergen in airway inflammation with an immune adjuvant. Substance P and CGRP are involved in the pathophysiological process of formaldehyde induced asthma. TRPA1 and TRPV1 channel proteins mediate the effect of gaseous formaldehyde to induce asthma and regulate neuropeptides. Release.
【学位授予单位】：华中师范大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R114;R562.25

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