miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡中的调控作用

目的探讨miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡和炎症反应中的调控作用。方法运用卵清蛋白(OVA)致敏的小鼠变应性鼻炎模型,使用实时定量PCR(qRT-PCR)、蛋白质印迹试验(Western Blot)、酶联免疫吸附试验(ELISA)、免疫组织化学检测鼻黏膜上皮细胞内miR-375、JAK2、细胞凋亡相关蛋白(JAK2蛋白,裂解的蛋白酶3(Cleaved caspase 3),聚[ADP-核糖]聚合酶裂解酶(Cleaved PARP),蛋白酶3(Caspase 3),聚[ADP-核糖]聚合酶(PARP),p-STAT3蛋白,STAT3蛋白和β肌动蛋白(β-actin))和血浆IL-6、TNF-α、IL-10的表达水平。结果miR-375在变应性鼻炎小鼠的鼻黏膜上皮细胞中表达降低,而JAK2表达增高;JAK2蛋白、p-STAT3蛋白和裂解的蛋白酶3均在OVA组表达增高;给OVA致敏的变应性鼻炎小鼠注射miR-375模拟物可以导致血清IL-6、TNF-α的分泌下降,而IL-10分泌增加,该作用可以被带有过表达JAK2的腺病毒感染后而减弱。结论miR-375/JAK2调控通路存在于变应性鼻炎鼻黏膜上皮细胞中,并通过JAK2/STAT3信号通路调控细胞的凋亡和炎症反应,miR-375在变应性鼻炎的病程中有保护性机制。

Regulation of miR-375 on apoptosis and inflammatory response in nasal mucosa of allergic rhinitis mice

Objective This study aims to investigate the role of miR-375 in the regulation of apoptosis and inflammatory response in nasal mucosal epithelial cells of allergic rhinitis mice.Method Allergic rhinitis mice model sensitized with ovalbumin(OVA),and using real-time quantitative PCR(qRT-PCR),Western Blot,enzyme-linked immunosorbent assay(ELISA),immunohistochemistry to detect miR-375,JAK2,apoptosis-related proteins(JAK2 protein、Cleaved caspase 3、Cleaved PARP,、Caspase 3、PARP、p-STAT3、STAT3 andβ-actin)and IL-6,TNF-α,IL-10 expression levels in plasma.Results MiR-375 expression was found to be decreased in nasal mucosal epithelial cells of allergic rhinitis mice,while JAK2、JAK2 protein、p-STAT3 protein and cleaved caspase 3 expression exhibited an opposite effect.Injection of miR-375 agomir following ovalbumin(OVA)treatment declined OVA-induced allergic rhinitis,characterized as the down-regulation of IL-6 and TNF-αsecretion,and up-regulation of IL-10 secretion,these effects were declined by infection with JAK2 adenovirus through nasal cavity inhalation.Conclusion miR-375/JAK2 regulatory pathway is identified in nasal mucosal epithelial cells of allergic rhinitis,and regulates apoptosis and inflammatory response through JAK2/STAT3 signaling pathway.MiR-375 has a protective mechanism in allergic rhinitis.