微RNA-214靶向性别决定区Y-box 4对类风湿关节炎滑膜成纤维细胞增殖凋亡的影响  

Effects of miR-214 on the proliferation and apoptosis of rheumatoid arthritis synovial fibroblasts by targeting sex determining region Y-box 4

作　　者：魏晓颖[1] 王军涛[1] 田景霞[2] 魏新平[1] Wei Xiaoying;Wang Juntao;Tian Jingxia;Wei Xinping(Nephrology and Rheumatology,First people's Hospital of Shangqiu,Henan 476100,China;Respiraiory and Critical Care Medicine,First Peopled Hospital of Shangqiu,Henan 476100,China)

机构地区：[1]河南省商丘市第一人民医院肾病风湿科,476100 [2]河南省商丘市第一人民医院呼吸与危重症医学科,476100

出　　处：《中华风湿病学杂志》2021年第7期455-460,I0002,共7页Chinese Journal of Rheumatology

摘　　要：目的探讨微RNA(miR)-214靶向性别决定区Y-box 4(SOX4)对RA滑膜成纤维细胞增殖、凋亡的影响。方法收集2017—2019年商丘市第一人民医院就诊的45例RA患者的滑膜组织样本和30例接受关节置换手术的关节创伤患者的滑膜组织样本,分别作为RA组和对照组。通过实时荧光定量PCR(RT-qPCR)检测滑膜组织中miR-214和SOX4表达水平;ELISA法检测患者血清RF、CRP浓度,采用血沉动态分析仪检测ESR浓度,Pearson相关分析miR-214及SOX4与患者血清RF、CRP、ESR相关性。采用Lipofectamine 3000将人成纤维样滑膜细胞(HFLS)-RA细胞分为对照组、miR-214 mimic组、mimic-NC组、miR-214 mimic+pcDNA3.1-SOX4组。通过CCK8检测各组细胞活力,流式细胞仪检测各组细胞凋亡情况,蛋白质印迹法检测相关指标蛋白表达水平。2组比较采用独立样本t检验,多组比较采用单因素方差分析,两两比较采用LSD-t检验。结果与对照组(4.6±0.7,1.9±0.7)和HFLS细胞(1.00±0.06,1.00±0.09)比,miR-214在RA患者组织(2.6±0.9)和HFLS-RA(0.30±0.05)中低表达(t=10.026,P<0.05;t=15.815,P<0.05);SOX4在RA患者组织(4.6±0.9)和HFLS-RA(3.89±0.41)中高表达(t=14.772,P<0.05;t=12.020,P<0.05)。RA组患者血清RF、ESR、CRP表达水平[(46±7)U/ml、(46.4±9.6)mm/1 h、(34.8±9.8)mg/ml]较对照组[(16±4)U/ml、(9.2±2.0)mm/1 h、(2.1±0.7)mg/ml]显著升高(t=22.906,P<0.05;t=25.338,P<0.05;t=22.314,P<0.05)。患者血清RF、CRP、ESR与miR-214呈负相关(r=-0.574,P<0.05;r=-0.448,P<0.05;r=-0.549,P<0.05),与SOX4、ESR呈正相关(r=0.492,P<0.05;r=0.369,P<0.05;r=0.325,P<0.05)。转染miR-214 mimic 24 h后细胞活力、Ki-67和p-p65/p65蛋白表达量较对照组和mimic-NC组显著下降(F_(24 h)=16.980、F_(48 h)=42.735、F_(72 h)=164.448、F_(Ki-67)=290.112、F_(p-p65/p65)=223.548,P<0.05),细胞凋亡率和Bax蛋白表达量显著升高(F=344.360,P<0.05;F=106.376,P<0.05)。过表达SOX4可逆转miR-214的作用。结论miR-214靶向SOX4抑制RA滑膜成纤维细胞增殖,并促进凋�Objective To investigate the effect of miR-214 targeting sex determining region Y-box 4(SOX4)on the proliferation and apoptosis of rheumatoid arthritis(RA)synovial fibroblasts.Methods Synovial tissue samples from 45 patients with rheumatoid arthritis and 30 patients with joint trauma who underwent joint replacement surgery from 2017 to 2019 in Shangqiu First People's Hospital samples were collected.The expression levels of miR-214 and SOX4 in synovial tissues were detected by real time quantitative polymerase chain reaction(RT-qPCR);The expression levels of serum rheumatoid factor(RF)and C-reactive protein(CRP)were detected by enzyme-linked immunosorbent assay(ELISA)test.The correlation between miR-214 or SOX4 and RF,CRP,erythrocyte sedimentation rate(ESR)was tested by Pearson's analysis.Human fibroblast-like synoviocytes(HFLS)-RA cells were derived from the control group,miR-214 mimic group,mimic-NC group and miR-214 mimic+pcDNA3.1-SOX4 group and se-parated by Lipofectamine 3000.The cell viability of each group was detected by CCK8,the apoptosis of each group was detected by flow cytometry,the target relationship was verified by dual luciferase experiment,Western-blot was used to detect the protein expression levels of related indicators.Independent sample t test was used for the comparison between the two groups,one-way nalysis of variance(ANOVA)was used for multi-group comparison,and LSD-t test was used for the comparison between the two groups.Results Compared with the control group(4.6±0.7,1.9±0.7)and HFLS cells(1.00±0.06,1.00±0.09),miR-214 was expressed lower in RA tissues(2.6±0.9)and HFLS-RA(0.30±0.05)(t=10.026,P<0.05;t=15.815,P<0.05).Compared with SOX4 was highly expressed in RA tissues(4.6±0.9)and HFLS-RA cells(3.89±0.41)(t=14.772,P<0.05;t=12.020,P<0.05).Serum RF,ESR and CRP expression levels in RA group[(46±7)U/ml,(46.4±9.6)mm/1 h,(34.8±9.8)mg/ml]were signifi-cantly higherthan those in the control group[(16±4)U/ml,(9.2±2.0)mm/1 h,(2.1±0.7)mg/ml](t=22.906,P<0.05;t=25.338,P<0.05;t=22.314,P<0

关 键 词：关节炎 类风湿 微RNAS 性别决定区Y蛋白质 核因子-ΚB信号通路 

分 类 号：R593.22[医药卫生—内科学]