LncRNA PVT1调控呼吸道合胞病毒感染人胚肺成纤维细胞凋亡的机制  

Influence of LncRNA PVT1 on apoptosis of human embryonic lung fibroblasts cells of patients with respiratory syncytial virus infection

作　　者：苏文雅 吴钰迪 邱慧 何骞 周琳 李志英 SU Wen-ya;WU Yu-di;QIU Hui;HE Qian;ZHOU Lin;LI Zhi-ying(The Third Affiliated Hospital of Soochow University(The First People's Hospital of Changzhou),Changzhou,Jiangsu 213004,China)

机构地区：[1]苏州大学附属第三医院(常州市第一人民医院)呼吸与危重症医学科,江苏常州213004

出　　处：《中华医院感染学杂志》2024年第13期1921-1927,共7页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金资助项目(81900058)。

摘　　要：目的分析长链非编码RNA(lncRNA)人浆细胞瘤转化迁移基因1(PVT1)对呼吸道合胞病毒(RSV)感染人胚肺成纤维(HELF)细胞损伤的影响。方法本研究纳入于2020年1月一2023年5月在常州市第一人民医院确诊并住院治疗的RSV感染的哮喘患者38例和健康志愿者38名。实时定量聚合酶链反应(qRT-PCR)检测lncRNAPVT1和miR-625-5p表达情况。将HELF细胞分为NC组(未感染)、RSV组(RSV感染)、si-NC+RSV组(转染si-NC+RSV感染)、si-lncRNAPVT1+RSV组(转染si-lncRNAPVT1+RSV感染)、miR-NC+RSV组(转染miR-NC+RSV感染)、miR-625-5p+RSV组(转染miR-625-5p模拟物+RSV感染)、anti-miR-NC+si-lncRNAPVT1+RSV组(共转染anti-miR-NC与si-lncRNAPVT1+RSV感染)、anti-miR-625-5p+si-lncRNA PVT1+RSV组(共转染anti-miR-625-5p与si-lncRNAPVT1+RSV感染)。采用流式细胞仪检测细胞凋亡,免疫印迹法检测蛋白表达,酶联免疫吸附测定法检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6含量。双荧光素酶报告实验分析lncRNAPVT1与miR-625-5p的靶向关系。结果RSV感染的患者血清和HELF细胞中lncRNAPVT1表达量比健康人血清或对照NC增加(均P<0.05)。RSV感染HELF增加Bax、Cleaved-caspase-3蛋白表达量、TNF-α、IL-6含量、凋亡率(均P<0.05);而这些影响在lncRNAPVT1沉默或miR-625-5p过表达后得到缓解(均P<0.05)。lncRNAPVT1靶向调控miR-625-5p的表达。anti-miR-625-5p+si-lncRNAPVT1+RSV组HELF细胞的Bax、Cleaved-caspase-3蛋白表达量、TNF-α、IL-6含量和细胞凋亡率均高于anti-miR-NC+si-lncRNAPVT1+RSV组(均P<0.05)。结论lncRNAPVT1低表达通过靶向miR-625-5p,抑制RSV感染的HELF细胞凋亡和炎症反应。OBJECTIVE To observe the influence of long-chain non-coding RNA(lncRNA)plasmacytoma variant translocation 1(PVT1)on damage of human embryonic lung fibroblasts(HELF)cells of the patients with respiratory syncytial virus(RSV).METHODS A total of 38 asthma patients with RSV infection who were hospitalized in the First Peoples Hospital of Changzhou from Jan 2020 to May 2023 were enrolled in the study,and 38 healthy volunteers were also enrolled in the study.The expressions of lncRNA,PVT1 and miR-625-5p were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The HELF cells were divided into the NC group(no infection),the RSV group(RSV infection),the si-NC+RSV group(transfected si-NC+RSV infection),the si-IncRNA PVT1+RSV group(transfection si-lncRNA PVT1+RSV infection),the miR-NC+RSV group(transfection miR-NC+RSV infection),the miR-625-5p+RSV group(transfection miR-625-5p mimic+RSV infection),the anti-miR-NC+si-lncRNA PVT1+RSV group(cotransfection of anti-miR-NC and si-lncRNA PVT1+RSV infection)and the anti-miR-625-5p+si-lncRNA PVT1+RSV group(cotransfection of anti-miR-625-5p and si-lncRNA PVT1+RSV).Cell apoptosis was detected by using flow cytometry,expressions of proteins were detected by Western Blot,the levels of tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 were detected by enzyme-linked immunosorbent assay.The dual luciferase report experiment was carried out to analyze the targeting relationship between the IncRNA PVT1 and the miR-625-5p.RESULTS The expression level of IncRNA PVT1 was higher in serum and HELF cells of the patients with RSV infection than in serum or control NC of the healthy people(all P<0.05).The expression levels of Bax,Cleaved-caspase-3 protein,levels of TNF-αand IL-6,and apoptosis rate of HELF were increased significantly among the patients with RSV infection(all P<O.05).All the effects were mitigated after the IncRNA PVT1 was silent or the miR-625-5p was overexpressed(all P<0.05).LncRNA PVT1 targeted and regulated the expression of miR-625-5p.The expression levels

关 键 词：哮喘 呼吸道合胞病毒 长链非编码RNA 人浆细胞瘤转化迁移基因1 miR-625-5p 人胚肺成纤维细胞 

分 类 号：R563.1[医药卫生—呼吸系统]