LncRNA PVT1 通过调节miR-214/STAT6 轴减轻哮喘模型小鼠气道炎性反应

doi:10.16352/j.issn.1001-6325.2022.09.1374

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (9): 1374-1380.doi: 10.16352/j.issn.1001-6325.2022.09.1374

LncRNA PVT1 通过调节miR-214/STAT6 轴减轻哮喘模型小鼠气道炎性反应

黄华¹, 周龙¹, 姚迪¹, 许诣^2*

1. 湖北省恩施土家族苗族自治州中心医院呼吸与危重症医学科, 湖北恩施 445000;
2. 湖北文理学院附属医院襄阳市中心医院儿科, 湖北襄阳 441021

收稿日期:2021-10-11 修回日期:2022-01-28 出版日期:2022-09-05 发布日期:2022-09-02
通讯作者: 15971039756@163.com
基金资助:
湖北省自然科学基金(WZ2017Q039)

LncRNA PVT1 alleviates inflammation of airway via regulating miR-214/STAT6 axis in mouse models with asthma

HUANG Hua¹, ZHOU Long¹, YAO Di¹, XU Yi^2*

1. Department of Respiratory and Critical Care Medicine, the Central Hospital of Hubei Enshi Tujia and Miao Autonomous Prefecture, Enshi 445000;
2. Department of Pediatrics, the Affiliated Hospital of Hubei University of Arts and Sciences, Xiangyang Central Hospital, Xiangyang 441021,China

Received:2021-10-11 Revised:2022-01-28 Online:2022-09-05 Published:2022-09-02

摘要/Abstract

摘要： 目的探究抑制lncRNA PVT1表达对支气管哮喘小鼠气道炎性反应的作用,以及对miR-214/STAT6轴的影响。方法将小鼠随机分成对照组、模型组[用卵清蛋白(OVA)诱导哮喘)]、lncRNA PVT1抑制组和lncRNA PVT1 NC组(尾静脉注射lncRNA PVT1抑制物或其阴性对照),每组12只。血细胞计数仪计数支气管肺泡灌洗液(BALF)中炎性细胞总数、巨噬细胞数、中性粒细胞数和淋巴细胞数;ELISA试剂盒检测BALF上清液中白介素-4(IL-4)、白介素-5(IL-5)、白介素-13(IL-13)浓度;试剂盒检测血清中IgE水平;HE染色观察肺组织炎性细胞浸润情况并进行评分;RT-qPCR检测肺组织中miR-214和STAT6 mRNA水平;Western blot检测肺组织中STAT6和p-STAT6蛋白表达;双荧光素酶报告基因检测分析lncRNA PVT1与miR-214靶向关系。结果与对照组相比,模型组小鼠BALF中炎性细胞总数、巨噬细胞数、中性粒细胞数和淋巴细胞数、IL-4、IL-5、IL-13水平、血清中IgE水平及肺组织中炎性细胞浸润程度评分、STAT6 mRNA和蛋白磷酸化水平均显著升高(P＜0.05),而肺组织中miR-214水平显著降低(P＜0.05)。与模型组和lncRNA PVT1 NC组相比,lncRNA PVT1抑制组BALF中炎性细胞总数、巨噬细胞数、中性粒细胞数和淋巴细胞数、IL-4、IL-5、IL-13水平、血清中IgE水平及肺组织中炎性细胞浸润程度评分、STAT6 mRNA和蛋白磷酸化水平均显著降低(P＜0.05),而肺组织中miR-214水平显著升高(P＜0.05)。miR-214与lncRNA PVT1具有明显的靶向关系(P＜0.05)。结论抑制lncRNA PVT1可能通过靶向提高miR-214表达水平,抑制STAT6磷酸化途径,减轻哮喘小鼠的气道炎性反应。

关键词: lncRNA PVT1, 哮喘, 气道炎性反应, miR-214/STAT6轴

Abstract: Objective To explore the effect of inhibiting the expression of lncRNA PVT1 on airway inflammation response and the effect on the miR-214/STAT6 axis in mice with bronchial asthma. Methods Mice were randomly divided into control group, model group [asthma induced by ovalbumin(OVA)], lncRNA PVT1 inhibitor group and lncRNA PVT1 NC group with 12 mice in each.Hemocytometer was used to count the total number of inflammatory cells, numbers of macrophages, neutrophils and lymphocytes in broncho-alveolar lavage fluid (BALF). ELISA kit was used to detect the concentration of interleukin-4(IL-4), interleukin-5 (IL-5) and interleukin-13 (IL-13) in the BALF supernatant. The kit was used to detect the level of IgE in serum. HE staining was used to observe and score the infiltration of inflammatory cells in lung tissue. RT-qPCR was used to detect the levels of miR-214 and STAT6 mRNA in lung tissue. Western blot was used to detect the expression of STAT6 and p-STAT6 proteins in lung tissue. The dual luciferase reporter gene detection was used to analyze the targeting relationship between lncRNA PVT1 and miR-214. Results Compared with the control group, the cell counting of inflammatory cells, macrophages, neutrophils and lymphocytes, level of IL-4, IL-5, IL-13 in BALF, serum IgE,inflammatory cell infiltration, STAT6 mRNA and protein phosphorylation in lung tissues of mice in the model group were all significantly increased (P＜0.05).While the miR-214 level in lung tissue was significantly reduced (P＜0.05). Compared with the model group and the lncRNA PVT1 NC group, the total number of inflammatory cells, macrophages, neutrophils and lymphocytes, IL-4, IL-5, IL-13 levels in BALF, and serum IgE ,inflammatory cell infiltration, STAT6 mRNA and protein phosphorylation in lung tissues in the lncRNA PVT1 inhibition group were all significantly reduced (P＜0.05). But the miR-214 exprssion in lung tissue was significantly increased(P＜0.05). miR-214 showed a significant targeting relationship with lncRNA PVT1 (P＜0.05). Conclusions Inhibiting lncRNA PVT1 may increase expression of miR-214 ,inhibit the STAT6 phosphorylation pathway and reduce airway inflammation response of mouse with asthma.

Key words: lncRNA PVT1, asthma, airway inflammation response, miR-214/STAT6 axis

中图分类号:

R726.1

黄华, 周龙, 姚迪, 许诣. LncRNA PVT1 通过调节miR-214/STAT6 轴减轻哮喘模型小鼠气道炎性反应[J]. 基础医学与临床, 2022, 42(9): 1374-1380.

HUANG Hua, ZHOU Long, YAO Di, XU Yi. LncRNA PVT1 alleviates inflammation of airway via regulating miR-214/STAT6 axis in mouse models with asthma[J]. Basic & Clinical Medicine, 2022, 42(9): 1374-1380.

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LncRNA PVT1 通过调节miR-214/STAT6 轴减轻哮喘模型小鼠气道炎性反应

LncRNA PVT1 alleviates inflammation of airway via regulating miR-214/STAT6 axis in mouse models with asthma

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