探讨芥菜籽对小鼠变态反应性皮肤病的预防作用

发布时间：2018-04-04 15:50

本文选题：芥菜籽　切入点：Ⅰ型变态反应　出处：《中国人民解放军医学院》2012年硕士论文

【摘要】：研究背景与目的近年来，随着全球工业化程度的不断提高，变态反应性疾病的发病率逐年升高，已经成为影响人类健康的主要疾病之一。其中，变态反应性皮肤病的发病率最高[1]。因此，，对于变态反应性皮肤病的研究和探讨有效的防治方法，目前已经成为医学研究的热点之一。目前治疗变态反应性皮肤病的方法主要是避免接触过敏原、药物治疗（抗组胺药、皮质类固醇激素）和特异性免疫治疗等，但不管采取哪种手段，变态反应性皮肤病的总体治愈率较低，患者长期的生活质量仍不理想。因而，变态反应性皮肤病的化学预防逐渐成为近年来研究的焦点。化学预防是应用天然产物和（或）人工合成的化学物质来逆转、抑制或预防疾病的发生、发展。目前通过膳食中存在的天然抗过敏成分来预防过敏是最有希望和潜力的方法。天然化学预防药物与人工合成的化学物质相比，更具安全性和实用性，也更易为人体所接受。流行病学研究发现，经常食用胡萝卜、金针菇、大枣、蜂蜜等食物能降低人类过敏性疾病的发病风险。芥菜籽（mustard seed，MS），又称芥子，是十字花科云苔属植物芥子的种子，其加工的产品芥末油和芥末粉是我们生活中常用的调味品。近年来研究发现芥菜籽及其活性提取物具有抗炎的作用[2]。变态反应实际上就是免疫特异性炎症反应。皮肤科临床最常见的是I和Ⅳ型变态反应，芥菜籽能否作为一种化学预防剂用于变态反应性皮肤病的预防，目前尚未见这方面的研究。基于以上背景，本实验建立小鼠皮肤局部毛细血管通透性增高模型、全身I型变态反应模型和Ⅳ型变态反应模型，初步探讨芥菜籽对小鼠变态反应性皮肤病的预防作用以及可能的作用机制，以期为芥菜籽在临床上的推广应用提供理论及实验依据。研究方法 1芥菜籽对皮肤I型变态反应的预防作用：①分别用组胺溶液和Compound48/80溶液建立小鼠皮肤毛细血管通透性增高的模型，采用分光光度计比色法测定芥菜籽对毛细血管通透性的影响。②用卵白蛋白（ovalbumin,OVA）建立小鼠全身I型变态反应模型，采用ELISA法测定芥菜籽对血清IFN-γ、IL-4及IgE含量的影响。 2芥菜籽对皮肤Ⅳ型变态反应的预防作用：①造模：用2.4-二硝基氟苯(DNFB)诱导小鼠变应性接触性皮炎（ACD）模型。②观察指标：耳部激发24h后，测量小鼠左右耳肿胀度和重量差，HE染色观察局部组织病理改变和炎症细胞浸润情况，免疫组化测量朗格汉斯细胞（LC）数量，RT-PCR测量局部促炎细胞因子TNF-α、IL-1β、IL-6mRNA表达，ELISA测血清TNF-ɑ因子的水平。结果 1芥菜籽对小鼠局部I型变态反应的影响：芥菜籽对组胺和Compound48/80诱发的小鼠毛细血管通透性增高均有抑制作用，芥菜籽组与对照组比较有显著差异（P0.05）。 2芥菜籽对小鼠全身I型变态反应的影响：血清中IgE、IL-4和IFN-γ的含量在模型组和空白组间有统计学差异（P0.05），模型组和芥菜籽组间无统计学差异（P0.05）。 3芥菜籽对小鼠变应性接触性皮炎的影响：芥菜籽能明显抑制ACD小鼠的耳肿胀度、降低左右耳重量差、减少局部炎症细胞浸润，降低组织局部IL-1β、IL-6和TNF-α的表达（P0.05），降低血清TNF-ɑ的含量（P0.05）。结论 1芥菜籽对组胺和Compound48/80溶液诱发的小鼠皮肤局部I型变态反应有明显抑制作用。 2芥菜籽对OVA诱发的小鼠全身I型变态反应血清中IgE、IL-4和IFN-γ的含量未见明显影响 3芥菜籽对DNFB诱发的小鼠变应性接触性皮炎的耳肿胀具有显著的抑制作用。 4芥菜籽对DNFB诱发的小鼠变应性接触性皮炎的抑制作用，可能与抑制LC细胞的抗原提呈、下调组织局部IL-1β、TNF-α、IL-6及血清中TNF-α等炎症细胞因子有关。 5研究开发芥菜籽具有十分广阔的应用前景。
[Abstract]:Research background and purpose
In recent years, with the increasing degree of global industrialization, allergic disease incidence increased year by year, has become one of the major diseases affecting human health. Among them, allergic skin disease has the highest incidence of [1]. therefore, for research of skin allergy and explore effective prevention methods, has become one of the focus of medical research.
The treatment of allergic skin disease is mainly to avoid contact with allergens, drug therapy (antihistamines, corticosteroids) and specific immunotherapy, but no matter what kind of means, the overall allergic skin disease cure rate is low, patients with long-term quality of life is still not ideal. Therefore, chemical the prevention of allergic skin disease has gradually become the focus of research in recent years. Chemical prevention is the application of natural products and (or) to reverse synthetic chemicals, occurrence, inhibition or prevention of disease development. The current through the presence of natural dietary anti allergic ingredients to prevent allergy is the most promising method and potential. Natural chemopreventive drug compared with synthetic chemicals, more safety and practicality, it is easily accepted by the human body.
Epidemiological studies have found, often eat carrots, mushroom, jujube, honey and other foods can reduce the risk of human allergic diseases. The mustard seed (mustard seed, MS), also known as mustard, cruciferous Brassica goes mustard seed and its processed products of mustard oil and mustard powder is a commonly used condiment we in life. Recent studies have found that [2]. allergic reaction of mustard seed and its extract has anti-inflammatory activity is actually immune specificinflammatory reaction. The most common clinical department of dermatology is I and type IV allergic reaction, mustard seeds can be used as a chemopreventive agent for the prevention of allergic skin diseases, there is no research.
Based on the above background, the experimental mice local skin capillary permeability model, body type I allergic reaction model and type IV allergic reaction model, preliminary study on preventive effect of mustard seed on mice allergic skin disease and the possible mechanism, to provide theoretical and experimental basis for the application of mustard seed in clinic..
research method
1 the preventive effect of mustard seed on the skin type I allergic reaction: histamine and Compound48/80 solution were used to establish mice skin capillary permeability increase model, using the spectrophotometer colorimetric determination of mustard seeds. The capillary permeability of ovalbumin (ovalbumin, OVA) to establish a mouse model of systemic allergy type I, determination of mustard seed on serum IFN- gamma using ELISA method. The effects of IL-4 and IgE content.
2 the preventive effect of mustard seed on skin type IV allergic reaction: model: 2.4- two 4-dinitrofluorobenzene (DNFB) induced allergic contact dermatitis (ACD) model. The observation indexes: ear excitation of 24h, measuring the left and right mouse ear swelling degree and the weight difference, observe the local pathological changes and inflammatory cells the infiltration of HE staining, immunohistochemical measurement (LC) Lange Hans cell number, RT-PCR measurement of local proinflammatory cytokines of TNF- alpha, IL-1 beta, IL-6mRNA expression, ELISA TNF- in serum was measured by alpha factor levels.
Result
1 mustard seed had a significant effect on local I type allergy in mice: Brassica juncea inhibited the increase of histamine and Compound48/80 induced capillary permeability in mice, and there was a significant difference between the mustard seed group and the control group (P0.05).
2 the effect of mustard seed on systemic I allergy in mice: there was a significant difference in serum IgE, IL-4 and IFN- gamma levels between the model group and the blank group (P0.05), but there was no significant difference between the model group and the mustard seed group (P0.05).
3 Effects of mustard seed on allergic contact dermatitis in mice: mustard seeds can inhibit ACD mice ear swelling, reduce left ear weight, reduce local infiltration of inflammatory cells, reduce the tissue expression of IL-6 and IL-1 beta, TNF- alpha (P0.05), and decrease the content of serum TNF- of the alpha (P0.05).
conclusion
1 mustard seeds have obvious inhibitory effect on the local I allergy of mice induced by histamine and Compound48/80 solution.
2 mustard seed has no significant effect on the content of IgE, IL-4 and IFN- gamma in the serum of OVA induced I type allergic reaction in mice.
3 mustard rapeseed has a significant inhibitory effect on the ear swelling of DNFB induced allergic contact dermatitis in mice.
The inhibitory effect of 4 mustard seed on DNFB induced allergic contact dermatitis in mice may be related to inhibiting the antigen presentation of LC cells and down regulating the inflammatory cytokines such as IL-1 TNF-, TNF- alpha, IL-6 and TNF- in the serum.
5 the research and development of mustard seed has a very broad application prospect.

【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R758.2

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