强直性脊柱炎模型小鼠踝关节组织和临床患者PBMC中miR-142-5p,SOCS1 mRNA表达及与免疫功能分析研究  

Expression of miR-142-5p,SOCS1 mRNA in Ankle Joint Tissue Spondylitis Model Mice and PBMC of Clinical Patients and Their Immune Function Analysis

作　　者：严鸣光[1] 方晓[1] 李汶轩 王可 殷卫兵[1] YAN Mingguang;FANG Xiao;LI Wenxuan;WANG Ke;YIN Weibing(Department of Clinical Laboratory,the First People’s Hospital of Shangqiu,Henan Shangqiu 476100,China)

机构地区：[1]商丘市第一人民医院检验科,河南商丘476100

出　　处：《现代检验医学杂志》2024年第3期29-36,共8页Journal of Modern Laboratory Medicine

基　　金：河南省科技攻关项目(资助单位:河南省科技厅,编号:212102310743):强直性脊柱炎环状RNA表达谱的研究及潜在分子;河南省医学科技攻关项目(资助单位:河南省卫生健康委员会,编号:2018020947):HLA-B27不同检测技术对强直性脊柱炎精准诊断的价值研究。

摘　　要：目的探究强直性脊柱炎(ankylosing spondylitis,AS)模型小鼠和临床患者外周血单个核细胞(peripheralblood mononuclear cell,PBMC)中微小RNA-142-5p(miR-142-5p),细胞因子信号转导抑制因子1(suppressor ofcytokine signaling 1,SOCS1)mRNA表达及其对免疫功能的影响。方法通过实时荧光定量(quantitative real timePCR,qRT-PCR)检测2022年1月~2023年3月商丘市第一人民医院收治的30例确诊的AS患者(患者组)和30例健康体检者(健康组)的PBMC中miR-142-5p和SOCS1 mRNA水平。采用牛蛋白聚糖联合完全弗氏佐剂诱导AS小鼠模型,然后将小鼠分为对照组、模型组、阴性组和拮抗剂组。对照组和模型组小鼠尾静脉注射生理盐水,阴性组和拮抗剂组小鼠分别尾静脉注射NC-antagomir和miR-142-5p-antagomir。治疗2周后,分别评估各组小鼠的关节炎症状评分。通过苏木精伊红(hematoxylin eosin,HE)染色评价踝关节形态。采用ELISA法检测小鼠血清中Th1细胞因子干扰素γ(IFN-γ)、Th2细胞因子白细胞介素-4(IL-4)、Th17细胞因子白细胞介素-17(IL-17)和Treg细胞因子叉头盒蛋白P3(FOXP3)的表达水平。通过qRT-PCR和Western blot检测PBMC和踝关节组织中miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17和FOXP3的mRNA和蛋白表达水平。结果与健康组比较,患者组PBMC中miR-142-5p水平升高(3.03±0.99 vs1.00±0.21),SOCS1 mRNA水平降低(0.41±0.09 vs 1.00±0.18),差异具有统计学意义(t=10.997,15.956,均P<0.001)。与对照组比较,模型组小鼠踝关节组织中miR-142-5p水平(4.00±0.52 vs 1.00±0.04)升高,IFN-γ和IL-17的mRNA和蛋白水平均升高,SOCS1,IL-4和FOXP3的mRNA和蛋白水平均降低,差异具有统计学意义(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,均P<0.05);关节炎症状评分升高(7.83±0.94 vs 0.00±0.00,t=22.212,P<0.05),踝关节结构破坏明显;血清中IFN-γ,IL-17水平以及IFN-γ/IL-4比值(0.81±0.08 vs 2.08±0.33)和IL-17/FOXP3比值(0.41±0.03 vs 1.27±0.10)均升高,�Objective To explore the expression of mocro RNA(miR)-142-5p and suppressor of cytokine signaling 1(SOCS1)mRNA in peripheral blood mononuclear cells(PBMC)of mice and clinical patients with ankylosing spondylitis(AS)and their impact on immune function.Methods The mRNA levels of miR-142-5p and SOCS1 in PBMC of 30 patients with AS(Patient group)and 30 healthy controls(Health group)treated in the First People’s Hospital of Shangqiu from January 2022 to March 2023 were measured by quantitative real time PCR(qRT-PCR).AS mice models were induced by bovine proteoglycan combined with complete Freund’s adjuvant,and these mice were divided into control group,model group,NC group and antagomir group.Normal saline was injected into tail vein in control group and model group,and NC-antagomir and miR-142-5p-antagomir were injected into tail vein in NC group and antagomir group,respectively.After 2 weeks of treatment,the arthritis symptom scores of mice in each group were evaluated.The morphology of ankle joint was evaluated by hematoxylin eosin(HE)staining.The levels of Th1 cytokine interferon-γ(IFN-γ),Th2 cytokine interleukin-4(IL-4),Th17 cytokine interleukin-17(IL-17)and Treg cytokine forkhead box protein P3(FOXP3)in PBMC of mice were detected by ELISA method.The mRNA and protein expression of miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17 and FOXP3 in PBMC and ankle joints were detected by qRTPCR and Western blot.Results Compared with Health group,the level of miR-142-5p in PBMC of patient group was increased(1.00±0.21 vs 3.03±0.99,t=10.997,P<0.001),while the level of SOCS1 mRNA was decreased(1.00±0.18 vs 0.41±0.09,t=15.956,P<0.001).Compared with control group,miR-142-5p level(1.00±0.04 vs 4.00±0.52)and the mRNA and protein levels of IFN-γand IL-17 in ankle joint tissue of model group were increased,while the mRNA and protein levels of SOCS1,IL-4 and FOXP3 were decreased,with significant differences(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,all P<0.05).The arthritis symptom score was inc

关 键 词：强直性脊柱炎 微小核糖核酸-142-5p 细胞因子信号转导抑制因子1 免疫功能 

分 类 号：R-332[医药卫生] R392.11