摘要
目的探讨转录因子SOX5在RA患者PBMCs及血清中的表达及其意义。方法招募南京医科大学第一附属医院风湿免疫科RA患者30例,OA患者27例和健康对照组(HC)30名。采用实时荧光定量(RT)-PCR检测SOX家族基因在RA患者PBMCs中相对表达量及3组PBMCs中SOX5表达,ELISA法检测3组血清中SOX5表达水平。运用RT-PCR检测分别使用IL-6、TNF-α、IL-1β及IL-17刺激PBMCs24h后,SOX5表达水平差异。免疫共沉淀实验(co-IP)法分析SOX5与核因子κB受体活化因子配体(RANKL)的相互作用。采用RNA干扰技术沉默小鼠破骨前体细胞系RAW264.7中SOX5表达,抗酒石酸酸性磷酸酶(TRAP)染色观察其对RANKL诱导破骨细胞分化的影响。多组间比较采用单因素方差分析,运用Pearson检验进行相关性分析。结果在检测的SOX基因家族8个基因中,RA患者PBMCs中以D亚家族的SOX5优势表达为主。RA组、OA组和健康对照组PBMCs中SOX5相对表达量分别为(21±19)、(10±8)、(5±4),RA组与OA组、健康对照组差异有统计学意义(F=8.343,P<0.01);RA组、OA组和健康对照组血清中SOX5平均水平分别为(19132±12054)pg/ml、(9065±15172)pg/ml和(3242±1251)pg/ml,RA组与OA组、健康对照组差异有统计学意义(F=15.31,P<0.01)。IL-6、TNF-α、IL-1β及IL-17可促进PBMCs中SOX5基因的表达。co-IP显示IL-6、TNF-α及IL-1β可促进SOX5与RANKL结合。采用SOX5-siRNA转染RAW264.7沉默SOX5表达后,RANKL诱导破骨细胞分化数量显著减少。结论RA患者PBMCs及血清中转录因子SOX5表达显著上调,可能通过调控RANKL基因表达,影响破骨细胞分化,参与RA发病过程。靶向抑制SOX5表达,可能成为治疗RA的新方法。
Objective To investigate the expression and clinical significance of transcription factor SOX5 in peripheral blood mononuclear cells (PBMCs) and serum in patients with rheumatoid arthritis (RA). Methods The relative expression of representative genes of the SOX gene family in the PBMCs from RA patients were detected by Real-Time Polymerase Chain Reaction(RT-PCR), and serum levels of SOX5 expression were measured by enzyme-linked immunosorbent assay (ELISA) in 30 RA patients, 27 osteoarthritis (OA) patients and 30 healthy controls (HC). The expression levels of SOX5 in PBMCs were detected by RT-PCR after stimulated with IL-6, TNF-α, IL-1β and IL-17 for 24 hours. The relationship between SOX5 and receptor activator of nuclear factor κB ligand (RANKL) was detected by co-Immunoprecipitation (co-IP). The formation of TRAP-positive cells after silence SOX5 in osteoclast precursor cell treated with RANKL was observed by Tartrate-resistant acid phasphate stain (TRAP). The differences were tested using one-way ANOVA followed by Student-Newman-Keuls post hoc analysis Correlations were analyzed using Pearson's analysis. Results SOX5 was predominantly expressed in the PBMCs of RA as compared with other SOX family genes in PBMC. PBMC levels of SOX5 in RA patients (21±19) were higher than the OA patients (10±8) and healthy control group (5±4)(F=8.343, P<0.01). While, Serum levels of SOX5 in RA patients [(19 132±12 054) pg/ml were higher than the OA patients [(9 065±15 172) pg/ml] and healthy control group [(3 242±1 251) pg/ml] (F=15.31, P<0.01). IL-6, TNF-α, IL-1β and IL-17 led to the up-regulation of SOX5 expression in PBMCs. IL-6, TNF-α, IL-1β promoted the interaction of SOX5 and RANKL in PBMCs. Silencing SOX5 reduced the formation of TRAP-positive cells in osteoclast precursor cell treated with RANKL. Conclusion Our results have proven that transcriptional factor SOX5 regulates the expression of RANKL and participates in the process of RA bone erosion. Inhibition of SOX5 expression may be a new therapy target o
作者
吴琴
石雨濛
骆爱姝
严伟
王芳
谈文峰
Wu Qin;Shi Yumeng;Luo Aishu;Yan Wei;Wang Fan;Tan Wenfeng(Department of Rheumatology,the First Affiliated Hospital of Nanjing Medical University,Jingsu 210029, China;Department of Cardiology,the First Affiliated Hospital of Nanjing Medical University,Jingsu 210020, China)
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2019年第1期46-51,共6页
Chinese Journal of Rheumatology
基金
国家自然科学基金(81471610,81172845,81471611,81671610).
