miR-302b-3p在变应性鼻炎小鼠模型中的作用与机制研究  

Study on the effect and mechanism of miR-302b-3p in an allergic rhinitis mouse model

作　　者：唐桥斐[1] 张爽[1] 闫智永[1] TANG Qiao-fei;ZHANG Shuang;YAN Zhi-yong(Department of Otolaryngology,the Second Affiliated Hospital of Shenyang Medical College,Shenyang 110002,China)

机构地区：[1]沈阳医学院附属第二医院耳鼻喉科,沈阳110002

出　　处：《现代免疫学》2022年第5期381-386,420,共7页Current Immunology

基　　金：辽宁省科学技术厅资助项目(2019JH8/10300055)。

摘　　要：为探讨miR-302b-3p对卵清蛋白(ovalbumin,OVA)所致变应性鼻炎(allergic rhinitis,AR)小鼠炎症反应的调控作用,将12只4周龄BALB/c小鼠随机分为4组:sham组、AR组、anti-miR-302b-3p NC组、anti-miR-302b-3p组。使用OVA制备AR模型,在建模后期,采用anti-miR-302b-3p NC或anti-miR-302b-3p抑制载体进行鼻内滴注。评估小鼠鼻部症状;采用H-E染色法观察鼻黏膜病变情况;qRT-PCR检测miR-302b-3p及B细胞淋巴瘤6(B-cell lymphoma 6,BCL6)mRNA水平;ELISA检测IL-4、IL-5及IL-13的水平;Western blotting检测BCL6、NOD样受体家族含pyrin结构域蛋白3(NOD-like receptor family pyrin domain-containing protein 3,NLRP3)、包含CARD的凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、前体Caspase-1(pro-Caspase-1)、活化的Caspase-1 p20亚基[cleaved-Caspase-1(p20)]及成熟IL-1β(mature-IL-1β)蛋白表达水平。结果显示,与sham组比较,AR组小鼠的鼻部过敏症状明显,鼻黏膜组织炎症反应较重,IL-4、IL-5及IL-13的分泌显著增加,NLRP3、ASC、cleaved-Caspase-1(p20)及mature-IL-1β的蛋白表达水平较高,miR-302b-3p表达增加,BCL6 mRNA及蛋白水平显著减少(均P<0.01);与anti-miR-302b-3p NC组比较,anti-miR-302b-3p组小鼠鼻部过敏症状减轻,鼻黏膜炎症反应减轻,上皮结构趋于完整,IL-4、IL-5及IL-13的分泌显著减少,NLRP3、ASC、cleaved-Caspase-1(p20)及mature-IL-1β的蛋白表达水平下降,miR-302b-3p表达减少,BCL6 mRNA及蛋白水平显著增加(均P<0.01)。该研究提示,miR-302b-3p可能通过靶向抑制BCL6调节NLRP3/ASC及下游炎症信号通路,进而促进AR进展。To study the regulatory effect of miR-302b-3p on the inflammatory response of allergic rhinitis(AR)mice induced by ovalbumin(OVA),this study included twelve BALB/c mice randomly divided into four groups:sham group,AR group,anti-miR-302b-3p NC group,and anti-miR-302b-3p group.The OVA-induced-AR model was established and in the later stage of model establishment,anti-miR-302b-3p NC or anti-miR-302b-3p inhibitory carrier was delivered by intranasal instillation.Toevaluate the nasal symptoms,H-E staining was used to reveal the pathological changes in the nasal mucosa.The expressions of miR-302b-3p and B-cell lymphoma 6(BCL6)mRNA were detected by qRT-PCR.The levels of IL-4,IL-5,and IL-13 were detected by ELISA.The expressions of BCL6,NOD-like receptor family pyrin domain-containing protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),pro-Caspase-1,cleaved-Caspase-1(p20),and mature-IL-1βwere detected by Western blotting.The results revealed that compared to the sham group,the AR group showed more obvious nasal allergy symptoms,more severe inflammation in the nasal mucosa,increased secretion of IL-4,IL-5,and IL-13,higher expression levels of NLRP3,ASC,cleaved-Caspase-1(p20),and mature-IL-1β,increased expression of miR-302b-3p,and increased mRNA and protein levels of BCL6(all P<0.01).Interestingly,compared to the anti-miR-302b-3p NC group,the nasal allergy symptoms of mice in the anti-miR-302b-3p group was significantly decreased.The results indicated that the nasal mucosal inflammatory response was reduced,the epithelial structure was less damaged,the secretion of IL-4,IL-5,and IL-13 decreased,and the protein levels of NLRP3,ASC,cleaved-Caspase-1(p20),and mature IL-1βdecreased,the expression of miR-302b-3p decreased,and the expression of BCL6 mRNA and protein increased(all P<0.01).The results suggest that miR-302b-3p may regulate NLRP3/ASC and the downstream inflammatory signaling pathway by targeted inhibition of BCL6,thus promoting AR progression.

关 键 词：变应性鼻炎 卵清蛋白 微小RNA-302b-3p B细胞淋巴瘤6 

分 类 号：R392.3[医药卫生—免疫学]