Department of Otorhinolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Department of Otorhinolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Int Immunopharmacol. 2023 May;118:110007. doi: 10.1016/j.intimp.2023.110007. Epub 2023 Mar 15.
MiR-223-3p is a multifunctional microRNA regulated by multiple transcription factors and plays a critical role in inflammation. This paper was designed to investigate the regulatory role and mechanism of miR-223-3p in eosinophils degranulation and allergic rhinitis (AR) inflammation.
OVA sensitized AR mouse model and EOL-1 cells model were established. RT-qPCR and FISH were performed to detect the miR-223-3p expression. ELISA and WB were utilized to evaluate mRNA and protein expression. HE staining and transmission electron microscopy were applied to observe the morphological changes in nasal mucosa. Flow cytometry and immunofluorescence staining were performed to measure the proportion of eosinophils and eosinophilic major basic protein expression. The targeting relationship between miR-223-3p and FBXW7 was verified by bioinformatic analysis and dual-luciferase reporter gene assay. The expression of FBXW7 was detected by immunohistochemistry.
The level of miR-223-3p in nasal mucosa was significantly up-regulated in AR group. The expression of miR-223-3p, ECP, MBP, and EPO were increased in EOL-1 cells, further increasing the miR-223-3p level could promote the ECP and EPO mRNA expression. Upregulation of miR-223-3p increased eosinophils granule protein expression, aggravated mucosal destruction and enhanced AR inflammation. Luciferase assay verified miR-223-3p directly target the 3'-UTR of FBXW7. In vitro, overexpression of FBXW7 could reverse the increase in MBP expression caused by the up-regulation of miR-223-3p. In vivo, knockdown of FBXW7 could reverse the down-regulation in granule protein level caused by the down-regulation of miR-223-3p, thereby aggravating AR inflammation.
Collected evidence elucidated that miR-223-3p could regulate the eosinophil degranulation and enhances the inflammation in AR by targeting FBXW7. The miR-223-3p/FBXW7 axis may provide a novel approach for AR treatment.
miR-223-3p 是一种受多种转录因子调控的多功能 microRNA，在炎症中发挥关键作用。本研究旨在探讨 miR-223-3p 在嗜酸性粒细胞脱颗粒和过敏性鼻炎（AR）炎症中的调节作用和机制。
建立 OVA 致敏 AR 小鼠模型和 EOL-1 细胞模型。采用 RT-qPCR 和 FISH 检测 miR-223-3p 的表达。ELISA 和 WB 用于评估 mRNA 和蛋白表达。HE 染色和透射电镜观察鼻黏膜形态变化。流式细胞术和免疫荧光染色检测嗜酸性粒细胞比例和嗜酸性粒细胞主要碱性蛋白表达。通过生物信息学分析和双荧光素酶报告基因检测验证 miR-223-3p 与 FBXW7 的靶向关系。免疫组化检测 FBXW7 的表达。
AR 组鼻黏膜 miR-223-3p 水平明显上调。EOL-1 细胞中 miR-223-3p、ECP、MBP 和 EPO 的表达增加，进一步提高 miR-223-3p 水平可促进 ECP 和 EPO mRNA 表达。上调 miR-223-3p 可增加嗜酸性粒细胞颗粒蛋白表达，加重黏膜破坏，增强 AR 炎症。荧光素酶检测证实 miR-223-3p 可直接靶向 FBXW7 的 3'-UTR。体外，过表达 FBXW7 可逆转 miR-223-3p 上调引起的 MBP 表达增加。体内，下调 miR-223-3p 可逆转 FBXW7 下调引起的颗粒蛋白水平降低，从而加重 AR 炎症。
研究结果表明，miR-223-3p 通过靶向 FBXW7 调节嗜酸性粒细胞脱颗粒并增强 AR 炎症。miR-223-3p/FBXW7 轴可能为 AR 治疗提供新的方法。
