鼻腔滴入乳铁蛋白防治小鼠变应性鼻炎及其机制的研究

发布时间：2018-10-31 11:16

【摘要】：目的：探讨乳铁蛋白在变应性鼻炎小鼠鼻腔中蛋白和基因表达水平及其和变应性鼻炎的关系,研究小鼠鼻腔滴入重组人乳铁蛋白对变应性鼻炎的防治作用,并探讨其可能的分子机制。方法：采用卵清蛋白建立BALB/c小鼠变应性鼻炎动物模型,用苏木素-伊红,高碘夫稀,肥大细胞染色方法分析鼻黏膜嗜酸性粒细胞,杯状细胞和肥大细胞浸润的情况。用荧光定量RT-PCR和酶联免疫方法检测小鼠鼻腔中T淋巴细胞亚群转录因子和细胞因子以及乳铁蛋白基因和蛋白的表达水平。用Spearman分析鼻腔中的乳铁蛋白表达水平和嗜酸性粒细胞的相关性。结果：与正常小鼠相比,嗜酸性粒细胞,杯状细胞和肥大细胞的数量,Th2、Th17、 Treg基因和蛋白表达水平在变应性鼻炎小鼠鼻腔显著增加(P0.01),但是在卵清蛋白激发前和激发后鼻腔给予重组人乳铁蛋白后均显著减少(P0.01)。Thl相关基因和蛋白表达水平在正常小鼠和模型小鼠之间无明显差异,但是给予重组人乳铁蛋白治疗后明显上调(P0.01)。与正常小鼠相比,内源性乳铁蛋白的基因和蛋白表达水平在变应性鼻炎小鼠鼻腔显著下调(P0.01),但是给予重组人乳铁蛋白治疗后明显上调(P0.01)。Spearman相关分析显示乳铁蛋白表达水平和嗜酸性粒细胞数量成负向关系(r=-0.920,P0.01)。结论：内源性乳铁蛋白在变应性鼻炎小鼠鼻腔中表达下调,该蛋白表达不足可能与变应性鼻炎的发生和发展相关。外源性乳铁蛋白抑制了小鼠变应性鼻炎的炎症,其机制可能与增强了内源性乳铁蛋白的表达,促进了Thl细胞活性和抑制Th2以及Th17细胞反应有关。该研究结果提示乳铁蛋白在未来可能作为一种新的预防和治疗变应性鼻炎的制剂。
[Abstract]:Objective: to investigate the expression of lactoferrin in nasal cavity of allergic rhinitis mice and its relationship with allergic rhinitis, and to study the preventive and therapeutic effects of recombinant human lactoferrin on allergic rhinitis. The possible molecular mechanism was also discussed. Methods: the animal model of allergic rhinitis in BALB/c mice was established by ovalbumin. The infiltration of eosinophils goblet cells and mast cells in nasal mucosa was analyzed by hematoxylin-eosin hyperiodophore and mast cell staining. The expression levels of transcription factors, cytokines, lactoferrin genes and proteins in T lymphocyte subsets in nasal cavity of mice were detected by fluorescence quantitative RT-PCR and enzyme-linked immunosorbent assay (Elisa). The correlation between the expression of lactoferrin and eosinophil in nasal cavity was analyzed by Spearman. Results: the number of eosinophilic granulocytes, goblet cells and mast cells, and the expression of Th2,Th17, Treg gene and protein were significantly increased in allergic rhinitis mice compared with normal mice (P0.01). However, the expression of recombinant human lactoferrin in nasal cavity before and after ovalbumin stimulation was significantly decreased (P0.01). Thl related gene and protein expression levels were not significantly different between normal mice and model mice. But after treatment with recombinant human lactoferrin, it was significantly up-regulated (P0.01). Compared with normal mice, the expression of endogenous lactoferrin gene and protein was significantly decreased in allergic rhinitis mice (P0.01). However, the expression of lactoferrin was significantly up-regulated after treatment with recombinant human lactoferrin (P0.01). Spearman correlation analysis showed that the expression of lactoferrin was negatively correlated with the number of eosinophils (r-0.920, P0.01). Conclusion: the expression of endogenous lactoferrin is down-regulated in the nasal cavity of allergic rhinitis mice, which may be related to the occurrence and development of allergic rhinitis. Exogenous lactoferrin inhibited the inflammation of allergic rhinitis in mice. The mechanism may be related to the enhancement of endogenous lactoferrin expression, the promotion of Thl cell activity and the inhibition of Th2 and Th17 cell response. The results suggest that lactoferrin may be a new preparation for the prevention and treatment of allergic rhinitis in the future.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R765.21

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