摘要
目的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(P<0.05),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(P<0.05)。Th1相关基因和蛋白表达水平在正常小鼠和模型小鼠之间差异无统计学意义,但是给予重组人乳铁蛋白治疗后明显上调(P<0.05)。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(P<0.05),但是给予重组人乳铁蛋白治疗后明显上调(P<0.05)。Spearman相关分析显示乳铁蛋白表达水平和嗜酸性粒细胞数量呈负相关(r=-0.920,P<0.05)。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Th1细胞活性和抑制Th2以及Th17细胞反应有关。
Objective To study the effect of intranasal administration recombinant human （rh） LF and its underlying mechanisms on AR in BALB/c mice. Methods BALB/c mice were used to establish allergic rhinitis which was induced by ovalbumin. The infiltration of eosinophils, goblet cells, and mast ceils in nasal mucosa was examined by hematoxylin＆eosin, periodic acid - Schiff, mast cell stai- ning, respectively. The expression levels of T - subsets related transcription factors and cytokines and lactoferrin mRNA and protein in the nasal mucosa were detected using real time RT - PCR and enzyme - linked immunosorbent assay among controls and allergic rhinitis, respectively. The method of Spearman was used to analyzed the relationship between lactoferrin expression and the number of eosinophils. Results We found that the number of eosinophils, goblet ceils, and mast ceils, as well as mRNA and protein expression of type 2 helper T （Th2）, Th17, and regulatory T （Treg） cells in the nasal cavity, were significantly up -regulated in AR mice compared with the controis （P 〈 0. 05 ）. Conversely, administration of rhLF prior to or after intranasal ovalbumin challenge markedly down - regulated these same parameters （P 〈 0.05 ）. Th1 - specific mRNA and protein expression in the nasal cavity of the controls was not different from that in AR mice, but expression significantly increased with rhLF treatment （ P 〈 0.05 ）. The mRNA and protein expression of endogenous LF in the nasal cavity was significantly down - regulated in AR mice compared with the controls （ P 〈 0.05 ）. However, after rhLF treatment, endogenous LF mRNA and protein expression was significantly upregulated （ P 〈 0. 05 ）. There was a negative correlation between the number of inflammatory ceils and lactoferrin expression levels （ r = - 0. 920, P 〈 0.05 ）. There was a negative correlation between the number of eosinophils and lactoferrin expression levels（ r = - 0. 920, P 〈 0.05）. Conclusion The expression levels of lacto
出处
《医学研究杂志》
2017年第3期157-162,共6页
Journal of Medical Research
