鼻腔滴注白细胞介素-22对小鼠变应性鼻炎的缓解作用

发布时间：2018-04-17 06:47

本文选题：鼻炎 + 变应性　；参考：《苏州大学》2013年硕士论文

【摘要】：目的：探讨鼻腔滴注重组小鼠白细胞介素 22（recombination murine interleukin 22/rm IL 22）对小鼠变应性鼻炎免疫应答的调节作用。方法：将24只6~8周的雌性BALB/c小鼠随机分为3组，每组8只，分别为对照组、模型组和IL 22组。模型组和IL 22组建立小鼠变应性鼻炎模型，同时IL 22组在每次激发前鼻腔滴注rm IL 22。首先应用间接免疫荧光法检测鼻黏膜上皮细胞是否表达IL 22受体（IL 22R1），最后一次激发后记录三组小鼠10分钟的抓挠数和喷嚏数，应用酶联免疫吸附法（enzyme linked immunosorbent assay, ELISA）检测外周血清中卵白蛋白特异性IgE（OVA sIgE）的浓度，，检测鼻腔灌洗液中干扰素 γ（IFN γ）、白细胞介素 4（IL 4）、白细胞介素 5（IL 5）、白细胞介素 10（IL 10）和胸腺基质淋巴细胞生成素（thymic stromal lymphopoietin, TSLP）的浓度，HE染色观察鼻黏膜局部嗜酸性粒细胞浸润情况。最后利用SPSS17.0统计学软件对数据进行统计学分析，以P0.05为差异有统计学意义。结果：IL 22R1在小鼠鼻黏膜上皮细胞表达，IL 22组较模型组小鼠变应性鼻炎抓挠和喷嚏症状减轻，鼻腔灌洗液中IL 4、IL 5和外周血清中OVA sIgE的浓度较模型组降低，局部嗜酸性粒细胞数侵润减少，三组间差异有统计学意义，鼻腔灌洗液中IFN γ、IL 10和TSLP的浓度在IL 22组和模型组间差异无统计学意义。结论：鼻腔滴注rm IL 22可以抑制变应性鼻炎小鼠的免疫应答，有望成为治疗变应性鼻炎的一种新方法。目的：探讨重组1、2和8型腺相关病毒（rAAV1、rAAV2和rAAV8）载体经小鼠鼻腔滴注转导增强绿色荧光蛋白（EGFP）基因在鼻黏膜上皮细胞的表达情况。方法：6只6~8周的雌性BALB/c小鼠随机分为3组，每组2只。用带有EGFP基因的重组1、2和8型腺相关病毒进行小鼠鼻腔滴注，第4周处死小鼠，获得小鼠完整鼻腔，固定脱钙后冰冻切片，然后在荧光显微镜下观察EGFP在三组小鼠鼻黏膜上皮细胞的表达情况。结果：rAAV1载体携带的EGFP成功转染，第4周稳定表达，另外两型重组腺相关病毒携带的EGFP均未见在小鼠鼻黏膜表达。结论：rAAV1载体可以转染小鼠鼻腔黏膜上皮细胞，并稳定表达携带的EGFP，可以作为鼻部疾病基因治疗的载体。
[Abstract]:Aim: to investigate the regulatory effect of 22(recombination murine interleukin 22/rm IL-22 on the immune response in mice with allergic rhinitis.Methods: Twenty-four female BALB/c mice aged 6 to 8 weeks were randomly divided into 3 groups: control group, model group and IL-22 group.Model group and IL-22 group were used to establish allergic rhinitis model in mice.The expression of IL-22 receptor IL-22R1 was detected by indirect immunofluorescence assay. After the last stimulation, the number of scratches and sneezes in the three groups were recorded for 10 minutes.The concentration of ovalbumin specific IgE(OVA in peripheral serum was detected by enzyme enzyme linked immunosorbent assay (Elica) by enzyme linked immunosorbent assay (Elisa).Detection of IFN- 纬, 4(IL, 5(IL and 10(IL in nasal lavage fluid and thymic stromal lymphopoietin (TSLP) in nasal lavage fluidGranulocyte infiltration.Finally, SPSS17.0 statistical software was used for statistical analysis of the data, with P0.05 as the difference was statistically significant.Results compared with the model group, the symptoms of scratching and sneezing of mice with allergic rhinitis were alleviated, and the concentrations of IL-4 and IL-5 in nasal lavage fluid and OVA sIgE in peripheral serum were lower in the control group than in the model group.The number of eosinophils in local eosinophils decreased, and there was significant difference among the three groups. There was no significant difference in the concentrations of IFN 纬 -IL-10 and TSLP in nasal lavage fluid between IL-22 group and model group.Conclusion: nasal drip of rm ~ + IL-22 can inhibit the immune response in mice with allergic rhinitis and may be a new method for the treatment of allergic rhinitis.Aim: to investigate the expression of EGFPgene of recombinant adeno-associated virus (rAAV1) rAAV2 and rAAV8 in nasal epithelial cells via nasal drip transduction in mice.Methods six female BALB/c mice aged 6 to 8 weeks were randomly divided into 3 groups with 2 mice in each group.Mice were given nasal drip with recombinant adeno-associated virus 1t2 and type 8 containing EGFP gene. The mice were killed at the 4th week. The complete nasal cavity of mice was obtained, and the frozen sections were fixed after decalcification.Then the expression of EGFP in nasal epithelial cells of three groups was observed under fluorescence microscope.Results the EGFP carried by 1: rAAV1 vector was successfully transfected and stably expressed at the 4th week. The EGFP of the other two types of recombinant adeno-associated virus was not expressed in the nasal mucosa of mice.ConclusionTwo one rAAV1 vector can be transfected into mouse nasal mucosal epithelial cells and stably express EGFP. it can be used as a gene therapy vector for nasal diseases.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R765.21

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