MicroRNA-466a-3p在小鼠变应性鼻炎中的作用及机制研究
发布时间:2018-09-13 15:18
【摘要】:背景:变应性鼻炎是耳鼻咽喉头颈外科中的常见疾病。它是由IgE介导的免疫反应,以Th2相关的细胞因子分泌增加,鼻黏膜嗜酸性粒细胞和鼻腔分泌物增多为特征的变态反应性疾病。 方法:本研究通过microRNA芯片技术筛选出变应性鼻炎小鼠鼻黏膜中表达发生显著性改变的microRNA.利用网站对相关microRNA靶点进行预测,结合当前文献研究的进展着眼于在变应性小鼠鼻黏膜中表达下降的mir-466a-3p,探讨其在小鼠变应性鼻炎发病过程中对Thl/Th2细胞失衡的影响。利用双荧光素酶的方法验证网站预测的GATA3是否确实为mir-466a-3p的靶基因。利用mir-466a-3p模拟物在小鼠脾单个核细胞过表达mir-466a-3p,检测GATA3蛋白的表达变化。另外,通过构建mir-466a-3p的模拟物,尾静脉注射变应性鼻炎小鼠,检测小鼠变应性鼻炎相关指标的改变及GATA3蛋白的表达变化。 结果:MicroRNA芯片结果发现,在变应性鼻炎小鼠鼻黏膜中部分microRNAs表达与正常小鼠差异显著。双荧光素酶报告系统证实GATA3为mir-466a-3p的靶基因,与网站预测的结果相符。同时,体外实验发现mir-466a-3p的表达增加可以抑制GATA3蛋白的表达;体内实验也发现给予变应性鼻炎小鼠mir-466a-3p模拟物后使该microRNA表达趋于正常,可以使GATA3蛋白的表达水平较变应性鼻炎组下调,从而起到调节Thl/Th2细胞的平衡,缓解变应性鼻炎症状,达到治疗变应性鼻炎的作用。 结论:本研究探究了microRNAs在变应性鼻炎发病中的作用,为其发病机制的研究提供全新的视角,为以mir-466a-3p作为分子靶点的变应性鼻炎的治疗奠定理论依据和实验参考,具有一定的理论与实践意义。
[Abstract]:Background: allergic rhinitis is a common disease in otolaryngology. It is an allergic disease mediated by IgE, characterized by increased secretion of cytokines associated with Th2, increased eosinophil and nasal secretion in nasal mucosa. Methods: in this study, microRNA microarray was used to screen microRNA. in nasal mucosa of allergic rhinitis mice. The site was used to predict the relevant microRNA targets, and the current research progress was focused on exploring the effect of mir-466a-3p, on the imbalance of Thl/Th2 cells in allergic rhinitis during the pathogenesis of allergic rhinitis in mice with decreased expression of mir-466a-3p, in the nasal mucosa of allergic mice. Double luciferase method was used to verify whether the GATA3 predicted by the website was indeed the target gene of mir-466a-3p. Mir-466a-3p mimics were used to detect the expression of GATA3 protein in murine splenic mononuclear cells by overexpression of mir-466a-3p,. In addition, by constructing the mimic of mir-466a-3p and injecting mice with allergic rhinitis via caudal vein, the changes of related indexes and the expression of GATA3 protein in allergic rhinitis mice were detected. Results the expression of microRNAs in nasal mucosa of allergic rhinitis mice was significantly different from that of normal mice. Double luciferase reporting system confirmed that GATA3 was the target gene of mir-466a-3p, which was consistent with the results predicted by the website. At the same time, it was found in vitro that the increased expression of mir-466a-3p could inhibit the expression of GATA3 protein, and the in vivo experiment also found that the expression of microRNA tended to normal after giving mir-466a-3p mimics of allergic rhinitis mice. The expression level of GATA3 protein was lower than that of allergic rhinitis group, which could regulate the balance of Thl/Th2 cells, alleviate the symptoms of allergic rhinitis, and achieve the effect of treating allergic rhinitis. Conclusion: this study explored the role of microRNAs in the pathogenesis of allergic rhinitis, provided a new perspective for the study of its pathogenesis, and laid a theoretical and experimental reference for the treatment of allergic rhinitis with mir-466a-3p as a molecular target. It has certain theoretical and practical significance.
【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
[Abstract]:Background: allergic rhinitis is a common disease in otolaryngology. It is an allergic disease mediated by IgE, characterized by increased secretion of cytokines associated with Th2, increased eosinophil and nasal secretion in nasal mucosa. Methods: in this study, microRNA microarray was used to screen microRNA. in nasal mucosa of allergic rhinitis mice. The site was used to predict the relevant microRNA targets, and the current research progress was focused on exploring the effect of mir-466a-3p, on the imbalance of Thl/Th2 cells in allergic rhinitis during the pathogenesis of allergic rhinitis in mice with decreased expression of mir-466a-3p, in the nasal mucosa of allergic mice. Double luciferase method was used to verify whether the GATA3 predicted by the website was indeed the target gene of mir-466a-3p. Mir-466a-3p mimics were used to detect the expression of GATA3 protein in murine splenic mononuclear cells by overexpression of mir-466a-3p,. In addition, by constructing the mimic of mir-466a-3p and injecting mice with allergic rhinitis via caudal vein, the changes of related indexes and the expression of GATA3 protein in allergic rhinitis mice were detected. Results the expression of microRNAs in nasal mucosa of allergic rhinitis mice was significantly different from that of normal mice. Double luciferase reporting system confirmed that GATA3 was the target gene of mir-466a-3p, which was consistent with the results predicted by the website. At the same time, it was found in vitro that the increased expression of mir-466a-3p could inhibit the expression of GATA3 protein, and the in vivo experiment also found that the expression of microRNA tended to normal after giving mir-466a-3p mimics of allergic rhinitis mice. The expression level of GATA3 protein was lower than that of allergic rhinitis group, which could regulate the balance of Thl/Th2 cells, alleviate the symptoms of allergic rhinitis, and achieve the effect of treating allergic rhinitis. Conclusion: this study explored the role of microRNAs in the pathogenesis of allergic rhinitis, provided a new perspective for the study of its pathogenesis, and laid a theoretical and experimental reference for the treatment of allergic rhinitis with mir-466a-3p as a molecular target. It has certain theoretical and practical significance.
【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
【共引文献】
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