微小RNA-411-3p在分泌性中耳炎表达及其对炎症的调控机制  

Expression of microRNA-411-3p in secretory otitis media and its regulatory mechanism

作　　者：邢金燕 刘鑫国 谭建成 赵慧贤 杨丹 Xing Jinyan;Liu Xinguo;Tan Jiancheng;Zhao Huixian;Yang Dan(Department of Otolaryngology,Zhumadian Central Hospital Affiliated to Huanghuai University,Zhumadian 463000,China)

机构地区：[1]黄淮学院附属驻马店市中心医院耳鼻喉头颈外科,463000

出　　处：《中华实验外科杂志》2022年第12期2417-2420,共4页Chinese Journal of Experimental Surgery

摘　　要：目的:观察微小RNA(miR)-411-3p在分泌性中耳炎表达并探讨其炎症的调控机制。方法:选取2019年7月到2021年7月黄淮学院附属驻马店市中心医院收治的74例中耳炎患者和74例健康体检者的血液样本作为研究对象,采用荧光定量聚合酶链反应(PCR)分析两组血清中miR-411表达水平,采用酶联免疫吸附实验(ELISA)分析两组血清炎性因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达水平。生物信息学和双荧光素酶报告基因分析miR-411-3p靶基因。蛋白质免疫印迹分析靶基因的表达水平。培养人中耳上皮细胞系(HMEEC),采用100 nmol/L脂多糖处理24 h,采用荧光定量PCR分析miR-411-3p和炎性因子水平。组间数据比较采用t检验。结果:对照组血清miR-411-3p表达水平(1.03±0.14)明显高于中耳炎组患者血清miR-411-3p表达水平(0.64±0.12),差异有统计学意义(t=17.250,P<0.05)。对照组血清TNF-α、IL-1β和IL-6水平[(27.19±6.25)、(29.84±7.07)、(23.67±5.62)pg/ml]明显高于中耳炎组患者血清TNF-α、IL-1β和IL-6水平[(101.69±11.03)、(95.68±5.86)、(67.80±11.91)pg/ml],差异有统计学意义(t=18.570、22.660、10.600,P<0.05)。中耳炎患者血清miR-411-3p表达水平与血清TNF-α、IL-1β和IL-6水平呈负相关(r=-0.677、-0.812、-0.612,P<0.05)。低氧诱导因子-1α(HIF-1α)是miR-411-3p的靶基因。miR-NC组细胞HIF-1α表达水平(1.21±0.21)明显高于miR-411-3p组细胞HIF-1α表达水平(0.48±0.11),差异有统计学意义(t=6.931,P<0.05)。脂多糖(LPS)处理的人中耳上皮细胞miR-411-3p表达水平(0.76±0.12)明显低于未处理细胞(1.07±0.09),差异有统计学意义(t=5.027,P<0.05)。LPS处理的人中耳上皮细胞HIF-1α表达水平(1.67±0.18)明显高于未处理细胞(0.97±0.15),差异有统计学意义(t=7.262,P<0.05)。结论:miR-411-3p在分泌性中耳炎患者中表达显著下调,通过介导HIF-1α促进炎性因子释放,进而促进中耳炎的发生Objective To investigate the expression level of microRNA(miR)-411-3p in secretory otitis media and its regulation mechanism.Methods The blood samples of 74 patients with otitis media and 74 healthy people admitted to our hospital from July 2019 to July 2021 were selected as the research objects.The expression level of miR-411 in the serum of the two groups was analyzed by fluorescent quantitative polymerase chain reaction(PCR),and the serum inflammatory factor interleukin-1β(IL-1β)、tumor necrosis factorα(TNF-α)And interleukin-6(IL-6)in the two groups was analyzed by enzyme-linked immunosorbent assay(ELISA).Bioinformatics and double luciferase reporter gene analysis of miR-411-3p target gene.The expression level of target gene was analyzed by Western blotting.Human middle ear epithelial cells(HMEEC)were cultured and treated with 100 nmol/L lipopolysaccharide for 24 hours.The levels of miR-411-3p and hypoxia inducible factor-1α(HIF-1α)were analyzed.T-test was used to compare the data between groups.Results The expression level of serum miR-411-3p in the control group(1.03±0.14)was significantly higher than that in the otitis media group(0.64±0.12)(t=17.250,P<0.05).serum TNF-α,IL-1βAnd IL-6 levels[(27.19±6.25),(29.84±7.07),(23.67±5.62)pg/ml]in control group were significantly higher than serum TNF-α,IL-1βAnd IL-6 levels[(101.69±11.03),(95.68±5.86),(67.80±11.91)pg/ml]in patients with otitis media(t=18.570,22.660,10.600,P<0.05).Expression of serum miR-411-3p and serum TNF-α,IL-1βand IL-6 in patients with otitis media was negatively correlated(r=-0.677,-0.812,-0.612,P<0.05).HIF-1αis the target gene of miR-411-3p.The expression level of HIF-1αin miR-NC group(1.21±0.21)was significantly higher than that in miR-411-3p group(0.48±0.11)(t=6.931,P<0.05).The expression level of miR-411-3p in LPS treated HMEEC(0.76±0.12)was significantly lower than that in untreated cells(1.07±0.09)(t=5.027,P<0.05).The expression level of HIF-1αin LPS treated HMEEC(1.67±0.18)was significantly higher than that in

关 键 词：微小RNA-411-3p 分泌性中耳炎 炎症 低氧诱导因子-1Α 

分 类 号：R764.21[医药卫生—耳鼻咽喉科]