microRNA-22-3p低表达上调幼年哮喘气道重塑大鼠的NK1R表达  

Low expression of microrna-22-3p up regulates NK1R expression in juvenile asthmatic airway remodeling rats

作　　者：宋建刚[1] 高永伟 刘庆 贾鲲鹏[1] 庞随军[1] 李元霞[1] SONG Jiangang;GAO Yongwei;LIU Qing;JIA Kunpeng;PANG Suijun;LI Yuanxia(Department of Pediatrics, The Affiliated Hospital of Yan'an University, Yan'an 716000, Shanxi, China)

机构地区：[1]延安大学附属医院儿科,陕西延安716000

出　　处：《西部医学》2021年第1期4-9,共6页Medical Journal of West China

基　　金：中华国际科学交流基金(Z2018SLXB009)。

摘　　要：目的探讨幼年哮喘气道重塑大鼠的microRNA-22-3p(miR-22-3p)表达水平和神经激肽受体1(NK1R)表达水平的相关性。方法将48只Wistar大鼠分为对照组、模型组、模型+mimic组、模型+mimic-NC组、模型+inhibitor组、模型+inhibitor-NC组。对照组注射生理盐水并吸入空气;模型组应用卵白蛋白(OVA)雾化吸入法建立哮喘大鼠模型;模型+mimic组、模型+mimic-NC组、模型+inhibitor组、模型+inhibitor-NC组,在大鼠每次激发前1 h内分别尾静脉注射1 mL的mimic、mimic-NC、inhibitor、inhibitor-NC。H&E染色法观察小鼠气道重塑变化,RT-qPCR检测miR-22-3p的水平变化。荧光素酶基因报告检测方法在大鼠气道平滑肌细胞RASMCs中验证miR-22-3p靶向NK1R。使用Western blot和免疫组织化学法检测NK1R的表达水平。结果与对照组比较,模型组发生明显的气道重塑现象,且气道组织中miR-22-3p水平降低(P<0.05)。在RASMCs细胞中,NK1R的3′UTR和miR-22-3p的直接结合,导致荧光素酶活性强度降低(P<0.05)。与对照组相比,模型组的NK1R的mRNA和蛋白的表达水平明显上调(P<0.01)。与模型+mimic-NC组比较,模型+mimic组miR-22-3p的水平上调,而NK1R的表达下调(均P<0.05)。与模型+inhibitor-NC组比较,模型+inhibitor组miR-22-3p的水平下调,而NK1R水平上调(均P<0.05)。结论哮喘诱发大鼠气道NK1R的表达增加,miR-22-3p直接靶向调节抑制哮喘大鼠气道重塑中NK1R的表达水平,下调miR-22-3p解除对NK1R的抑制从而上调NK1R水平。Objective To investigate whether the expression level of microRNA-22-3p(miR-22-3p)and the neurokinin receptor 1(NK1R)in juvenile asthmatic airway remodeling rats are affected by the expression level of miR-22-3p.Methods Forty eight Wistar rats were divided into control group,model group,model+mimic group,model+mimic NC group,model+inhibitor group and model+inhibitor NC group.The control group was injected with normal saline and inhaled air;the model group was established with ovalbumin(OVA)atomization inhalation method;the model+mimic group,model+mimic NC group,model+inhibitor group and model+inhibitor NC group were injected with 1 mL of mimic,mimic NC,inhibitor and inhibitor NC by tail vein within 1 hour before each challenge.The changes of airway remodeling were observed by H&E staining and Mir-22-3p was detected by RT-qPCR.Luciferase gene reporter assay was used to verify that Mir-22-3p targeted NK1R in rat airway smooth muscle cells(RASMCs).The expression of NK1R was detected by Western blot and immunohistochemistry.Results Compared with the control group,significant airway remodeling was observed in the model group,and the level of Mir-22-3p in the airway tissue was decreased(P<0.05).In RASMCs,the direct binding of 3′-UTR and Mir-22-3p of NK1R resulted in the decrease of luciferase activity(P<0.05).Compared with the control group,the expression of NK1R mRNA and protein in the model group was significantly up-regulated(P<0.01).Compared with model+mimic NC group,the expression of Mir-22-3p in model+mimic group was up-regulated,while the expression of NK1R was down regulated(P<0.05).Compared with model+inhibitor NC group,the level of Mir-22-3p was down regulated in model+inhibitor NC group,while NK1R level was up-regulated(P<0.05).Conclusion The expression of NK1R in airway of asthmatic rats was increased.Mir-22-3p could directly target and inhibit the expression of NK1R in airway remodeling of asthmatic rats,and down regulate Mir-22-3p to relieve the inhibition of NK1R,thus increasing the level of NK1R.

关 键 词：microRNA-22-3p 幼年大鼠 哮喘 神经激肽受体1 气道平滑肌重塑 

分 类 号：R562.2+5[医药卫生—呼吸系统]