LINC00837/miR-671-5p/SERPINE2功能轴促进类风湿关节炎成纤维细胞样滑膜细胞的恶性病理学过程  

LINC00837/miR-671-5p/SERPINE2 functional axis promotes pathological processes of fibroblast-like synovial cells in rheumatoid arthritis

作　　者：曹周芳 汪元[2] 王梦娜 孙玥[2] 刘菲菲 CAO Zhoufang;WANG Yuan;WANG Mengna;SUN Yue;LIU Feifei(First Clinical Medical College,Anhui University of Traditional Chinese Medicine,Hefei 230038,China;First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学第一临床医学院,安徽合肥230038 [2]安徽中医药大学第一附属医院风湿免疫科,安徽合肥230031

出　　处：《南方医科大学学报》2025年第2期371-378,共8页Journal of Southern Medical University

基　　金：安徽省中医药传承创新科研项目(2024CCCX105);安徽省临床医学研究转化专项项目(202304295107020110);安徽省自然基金面上项目(2208085MH268);安徽省高等学校科学研究重点项目(2023AH050810)。

摘　　要：目的探讨LINC00837/miR-671-5p/SERPINE2功能轴对类风湿关节炎成纤维样滑膜细胞(RA-FLS)病理学过程的影响。方法选用正常成纤维细胞样滑膜细胞(FLS)和RA-FLS,构建LINC00837的过表达质粒和干扰质粒转染至RA-FLS中,实验分为对照组(FLS)、模型组(RA-FLS)、pcDNA3.1-NC组、pcDNA3.1-LINC00837组、siRNA-NC组、siRNA-LINC00837组。采用双荧光素酶验证LINC00837靶向miR-671-5p及miR-671-5p靶向SERPINE2关系;使用qRT-PCR实验检测各组细胞LINC00837、miR-671-5p、SERPINE2表达量;Edu法检测细胞增殖情况,划痕愈合实验检测RA-FLS迁移数,Western blotting法检测细胞增殖、迁移相关蛋白表达水平;ELISA法检测炎性细胞因子分泌情况。结果双荧光素酶报告基因结果显示,LINC00837与miR-671-5p的3'非翻译区(3'UTR)结合,而miR-671-5p与SERPINE2的3'非翻译区(3'UTR)结合,两两之间具有结合位点(P<0.01)。同一时间段,与对照组相比,模型组LINC00837和SERPINE2表达量升高,而miR-671-5p表达量降低,细胞增殖、迁移能力提高,促炎细胞因子表达升高,抑炎细胞因子表达降低(P<0.01);与模型组相比,pcDNA-LINC00837组细胞增殖、迁移活跃,促炎细胞因子表达升高,抑炎细胞因子表达降低(P<0.01);与模型组相比,siRNA-LINC00837组细胞增殖、迁移能力下降,促炎细胞因子表达下降,抑炎细胞因子水平升高(P<0.01)。结论RA-FLS中存在LINC00837/miR-671-5p/SERPINE2功能轴,该功能轴促进RA-FLS增殖、迁移及炎症因子分泌等恶性病理学过程,干预LINC00837有望成为调控RA-FLS病理学过程的潜在策略。Objective To investigate the regulatory effect of LINC00837/miR-671-5p/SERPINE2 functional axis on pathological processes of fibroblast-like synovial cells(FLS)in rheumatoid arthritis(RA).Methods RA-FLS were transfected with a LINC00837 overexpression plasmid(pcDNA3.1-LINC00837),a LINC00837 interference plasmid(siRNA-LINC00837),or their respective negative control plasmids(pcDNA3.1-NC and siRNA-NC).Dual luciferase was used to verify the targeting relationship between LINC00837 and miR-671-5p and between miR-671-5p and SERPINE2.RT-qPCR was used to detect the expression levels of LINC00837,miR-671-5p and SERPINE2 in normal FLS or the transfected cells,whose proliferation and migration abilities were assessed using Edu assay and scratch healing assay and by detecting the expression levels of Ki-67,PCNA,E-cadherin and N-cadherin with Western blotting.The changes in cellular secretion of the inflammatory cytokines(TNF-α,IL-17,IL-4 and IL-10)were examined using ELISA.Results Dual luciferase reporter gene assay showed that LINC00837 was capable of binding to the 3'-UTR of miR-671-5p,and the latter bound to the 3-UTR of SERPINE2 at specific binding sites between them.Compared with normal FLS,RA-FLS showed significantly increased expressions of LINC00837 and SERPINE2,lowered miR-671-5p expression and enhanced proliferation and migration abilities with increased expressions of pro-inflammatory cytokines and reduced expressions of anti-inflammatory cytokines.Transfection of RA-FLS with pcDNA-LINC00837 further enhanced cell proliferation and migration and the changes in the inflammatory cytokines,while transfection with si-LINC00837 produced the opposite changes.Conclusion RA-FLS have a LINC00837/miR-671-5p/SERPINE2 functional axis,which regulates cell proliferation,migration and secretion of inflammatory factors,and interventions targeting LINC00837 may provide a potential strategy to regulate the pathological processes in RA-FLS.

关 键 词：类风湿关节炎 成纤维细胞样滑膜细胞 LINC00837 增殖迁移 炎症因子 

分 类 号：R593.22[医药卫生—内科学]