长链非编码RNA NEAT1通过调节miR-221-3p对哮喘炎症和细胞屏障损伤的影响  

Effect of long non-coding RNA NEAT1 in asthmatic inflammation and cell barrier damage by regulating miR-221-3p

作　　者：王东 王璐璐 WANG Dong;WANG Lu-lu(Department of Pulmonary Disease,Henan Province Hospital of Traditional Chinese Medicine,Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan Province,China)

机构地区：[1]河南省中医院、河南中医药大学第二附属医院肺病科,河南郑州450000

出　　处：《中国临床药理学杂志》2022年第6期495-498,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究长链非编码RNA NEAT1在哮喘中的作用及其相关机制。方法将人气道平滑肌细胞(ASMCs)分为对照组(常规培养,不做处理),模型组[用血小板活化因子(PAF)处理ASMCs 12 h],转染组1(转染空载质粒24 h后,给予PAF处理12 h),转染组2(转染NEAT1过表达质粒24 h后,给予PAF处理12 h)。通过实时定量逆转录聚合酶链反应(RT-qPCR)法检测NEAT1和微小RNA-221-3p(miR-221-3p)的表达。用酶联免疫吸附(ELISA)法检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的浓度;比较各组ASMCs之间的单层通透性;用生物信息学和双荧光素酶报告基因实验验证NEAT1和miR-221-3p之间的相关性。结果对照组、模型组、转染组1和转染组2的ASMCs中NEAT1 mRNA的表达水平分别为1.00±0.00,1.54±0.07,1.50±0.13和2.12±0.13;miR-221-3p的表达水平分别为1.00±0.00,0.55±0.02,0.62±0.04和0.34±0.03;IL-1β分别为(1.57±0.09),(6.48±0.37),(6.16±0.55)和(15.21±0.98)pg·mL^(-1);IL-6分别为(153.51±8.37),(753.41±49.86),(712.79±83.83)和(1179.52±85.86)pg·mL^(-1);TNF-α分别为(9.94±0.56),(28.33±1.13),(26.52±0.90)和(54.86±3.74)pg·mL^(-1);ASMCs的单层通过率分别为(4.44±0.21)%,(8.43±0.39)%,(8.27±0.47)%和(16.54±0.78)%。对照组与模型组,转染组1和转染组2相比,差异均有统计学意义(均P<0.05)。生物信息学和双荧光素酶报告基因检测实验结果表明NEAT1可以靶向调控miR-221-3p(P<0.05)。结论NEAT1通过靶向miR-221-3p加重了哮喘气道炎症和细胞屏障损伤。Objective This study aimed to explore the role of long non-coding RNA NEAT1 in asthma and the related mechanism.MethodsAirway smooth muscle cells (ASMCs) were divided into control group(routinely cultured;without treatment),model group[ASMCs treated with platelet activating factor (PAF) for 12 h],transfection-1 group(treated with PAF for 12 h,at 24 h after the transfection of empty plasmids),and transfection-2 group (24 h after the transfection of NEAT1 overexpression plasmids,PAF treatment was given for 12 h).Real-time quantitative polymerase chain reaction (RT-q PCR) was used to detect the expression of NEAT1 mRNA and microRNA-221-3p(miR-221-3p).The levels of interleukin-1β(IL-1β),interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA).The monolayer permeability was compared between the ASMCs of each group.Bioinformatics and dual-luciferase reporter gene assay were used to verify the correlation between NEAT1 and miR-221-3p.Results The expression levels of NEAT1 mRNA in the ASMCs of the control group,model group,transfection-1 group and transfection-2 group were 1.00±0.00,1.54±0.07,1.50±0.13 and 2.12±0.13,respectively;the expression levels of miR-221-3p were 1.00±0.00,0.55±0.02,0.62±0.04 and 0.34±0.03;IL-1βwere (1.57±0.09),(6.48±0.37),(6.16±0.55) and(15.21±0.98) pg·m L^(-1);IL-6 were (153.51±8.37),(753.41±49.86),(712.79±83.83) and(1 179.52±85.86) pg·m L^(-1);TNF-αwere (9.94±0.56),(28.33±1.13),(26.52±0.90) and(54.86±3.74) pg·m L^(-1);the momolayer permeability rates of ASMCs were (4.44±0.21)%,(8.43±0.39)%,(8.27±0.47)%and (16.54±0.78)%,respectively.The differences between the control group and the model group,as well as between the transfection-1 group and the transfection-2 group were all statistically significant (all P<0.05).The results of bioinformatics and dual-luciferase reporter gene detection assay showed that NEAT1 could target and regulate miR-221-3p (P<0.05).Conclusion NEAT1 aggravates airway inflammation and ce

关 键 词：核旁斑装配转录本1(NEAT1) 微小RNA221-3p(miR-221-3p) 哮喘 气道炎症 细胞屏障损伤 

分 类 号：R97[医药卫生—药品]