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。
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
作者
黄华
周龙
姚迪
许诣
HUANG Hua;ZHOU Long;YAO Di;XU Yi(Department of Respiratory and Critical Care Medicine,the Central Hospital of Hubei Enshi Tujia and Miao Autonomous Prefecture,Enshi 445000;Department of Pediatrics,the Affiliated Hospital of Hubei University of Arts and Sciences,Xiangyang Central Hospital,Xiangyang 441021,China)
出处
《基础医学与临床》
2022年第9期1374-1380,共7页
Basic and Clinical Medicine
基金
湖北省自然科学基金(WZ2017Q039)。
