胸腺基质淋巴细胞生成素在特应性皮炎小鼠模型中的实验研究

发布时间：2018-06-08 04:35

本文选题：特应性皮炎 + TSLP　；参考：《泸州医学院》2012年硕士论文

【摘要】：目的：通过建立小鼠特应性皮炎模型,检测胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin, TSLP)在小鼠皮损及血清中的表达以及白介素-4(interleukin-4, IL-4)、γ干扰素(interferon-γ,IFN-γ)和免疫球蛋白E (immunoglobulin-E, IgE)在小鼠血清中的表达水平,分析TSLP与IL-4、IFN-γ和IgE的血清表达水平的相关性。探讨TSLP在特应性皮炎发病机制中的作用。方法：采用随机对照实验方法将18只BALB/c小鼠随机分入实验组和对照组。实验组用2.4-二硝基氟苯(2,4-Dinitrofluorobenzene, DNFB)丙酮/橄榄油(3：1)溶液致敏与激发建立小鼠特应性皮炎模型；对照组仅用丙酮/橄榄油(3：1)溶液。用苏木素-伊红(HE)染色方法观察两组小鼠皮损病理变化,并采用免疫组化技术检测TSLP在两组小鼠皮损组织中的表达,用酶联免疫吸附试验(Enzyme-linked immunosorbent assay, ELISA)方法检测两组小鼠血清中TSLP、IL-4、IFN-γ及IgE的表达水平。用SPSS17.0统计软件包对资料进行统计分析,计量资料比较用t检验,指标间的相关性用spearman直线相关分析。结果：1.小鼠皮损表现：实验组小鼠皮肤可见红斑、丘疹、结痂、苔藓样变等皮损改变；对照组小鼠皮肤表现正常。2.皮损组织学变化：2.1皮损组织病理表现：实验组见表皮轻度角化过度伴角化不全,棘层肥厚,棘细胞间海绵水肿,真皮浅层以淋巴细胞为主的慢性炎细胞浸润；对照组组织结构基本正常,未见明显炎性细胞浸润。2.2皮损组织免疫组化表现：TSLP阳性表达物质主要位于胞核,胞浆及胞膜无特异性着色,实验组平均光密度值(74.306±17.898)明显高于对照组(46.093±3.347),差异有统计学意义(p0.05)。3.TSLP、IL-4、IFN-γ和IgE血清学水平比较：3.1 TSLP：实验组血清TSLP水平为213.621 ± 34.666pg/ml,对照组血清TSLP水平为151.997±22.328pg/m1,实验组血清TSLP水平显著高于对照组,差异有统计学意义(p0.05)。3.2IL-4：实验组血清IL-4水平为97.831±15.882pg/ml,对照组血清IL-4水平为60.56l±9.286pg/ml,实验组血清工L-4水平显著高于对照组,差异有统计学意义(p0.05)。3.3 IFN-γ：实验组血清IFN-γ水平为70.999±9.064pg/ml,对照组血清IFN-γ水平为51.224±4.534 pg/ml,实验组血清IFN-γ水平显著高于对照组,差异有统计学意义(p0.05)。3.4 IgE：实验组血清IgE水平为142.488±16.352ng/m1,对照组血清IgE水平为14.851±3.971ng/ml,实验组工gE水平显著高于对照组,差异有统计学意义(p0.05)。4.实验组血清TSLP与IL-4、IFN-γ及IgE相关性分析：实验组TSLP血清水平与IL-4及IgE血清水平呈正相关(p0.05),而与IFN-γ血清水平呈负相关(p0.05)。结论：1.采用DNFB致敏和激发小鼠皮肤表面能够成功建立小鼠特应性皮炎模型。2.TSLP在特应性皮炎小鼠皮损组织及血清中大量表达,TSLP可促进IL-4及IgE的表达,而抑制IFN-γ的表达,其在特应性皮炎的发病机制中可能起着重要作用。3.特应性皮炎小鼠血清中IL-4、IFN-γ及IgE水平升高,这些细胞因子在特应性皮炎的发病过程中可能扮演着重要角色。
[Abstract]:Objective: to determine the expression of thymic stromal lymphopoietin (TSLP) in the skin lesions and serum of mice, and the expression of interleukins -4 (interleukin-4, IL-4), interferon (interferon- y, IFN- gamma) and immunoglobulin E (immunoglobulin-E, IgE) in the mice serum by establishing a mouse atopic dermatitis model. The correlation between TSLP and the serum levels of IL-4, IFN- gamma and IgE was analyzed. The role of TSLP in the pathogenesis of atopic dermatitis was investigated. Methods: 18 BALB/c mice were randomly divided into experimental and control groups by random control experiment. The experimental group was treated with 2.4- two nitrofluoro (2,4-Dinitrofluorobenzene, DNFB) acetone / olive oil. (3:1) a mouse atopic dermatitis model was established by solution sensitization and stimulation; the control group only used acetone / olive oil (3:1) solution. The pathological changes of skin lesions in two groups of mice were observed with hematoxylin eosin (HE) staining, and the expression of TSLP in the skin lesions of two groups of mice was detected by immunohistochemical technique, and the enzyme linked immunosorbent assay (Enzyme-linked Immunosorbent assay, ELISA) method was used to detect the expression level of TSLP, IL-4, IFN- gamma and IgE in the serum of two groups of mice. The data were statistically analyzed with SPSS17.0 statistical package, the measurement data were compared with t test, and the correlation between the indexes was analyzed by Spearman linear correlation. Results: 1. mice skin lesions showed red spots in the skin of the experimental mice. Changes in skin lesions such as papules, scab and moss like changes in the skin of the control group; the pathological changes of normal.2. skin lesions in the control group of mice: 2.1 lesions in the skin lesions: mild keratinization and keratinization in the experimental group, spinous cell hypertrophy, spongedema between spinous cells, and chronic lymphocytic infiltration of superficial lymphoblastic cells in the superficial layer of the dermis; the control group. The structure was normal and no obvious inflammatory cell infiltration of.2.2 skin tissue was found. The positive expression of TSLP was mainly located in the nucleus, and the cytoplasm and membrane had no specific coloring. The average optical density (74.306 + 17.898) of the experimental group was significantly higher than that of the control group (46.093 + 3.347). The difference was statistically significant (P0.05).3.TSLP, IL-4, IFN- gamma and IgE Serological comparison: 3.1 TSLP: the level of serum TSLP in the experimental group was 213.621 + 34.666pg/ml, and the level of serum TSLP in the control group was 151.997 + 22.328pg/m1. The serum TSLP level of the experimental group was significantly higher than that of the control group. The difference was statistically significant (P0.05).3.2IL-4: the serum IL-4 water level was 97.831 + 15.882pg/ml in the experimental group, and the serum IL-4 level in the control group. For 60.56l + 9.286pg/ml, the serum level of L-4 in the experimental group was significantly higher than that of the control group. The difference was statistically significant (P0.05).3.3 IFN- Gamma: the serum IFN- gamma level was 70.999 + 9.064pg/ml in the experimental group and the level of IFN- gamma in the control group was 51.224 + 4.534 pg/ml. The serum IFN- gamma level in the experimental group was significantly higher than that in the control group. The difference was statistically significant (P0.05). IgE: the level of serum IgE in the experimental group was 142.488 + 16.352ng/m1, the level of serum IgE in the control group was 14.851 + 3.971ng/ml, and the level of gE in the experimental group was significantly higher than that of the control group. The difference was statistically significant (P0.05) in the experimental group (P0.05), the serum TSLP and IL-4, IFN- gamma and IgE correlation analysis: the serum level of the experimental group was positively correlated with the serum level and the serum level. .05) and negative correlation with IFN- gamma serum level (P0.05). Conclusion: 1. the use of DNFB sensitization and stimulation of mouse skin surface can successfully establish a mouse atopic dermatitis model.2.TSLP in the skin tissue and serum of atopic dermatitis mice, TSLP can promote the expression of IL-4 and IgE, and inhibit the expression of IFN- gamma, and it is in atopic dermatitis. The pathogenesis of.3. atopic dermatitis may play an important role in the pathogenesis of atopic dermatitis, which may play an important role in the serum levels of IL-4, IFN- gamma and IgE in the serum of mice with atopic dermatitis.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R758.2;R-332

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