CAJ | 학술논문

目的本研究通过蛋白质组学的方法比较类风湿关节炎(RA)、骨关节炎及强直性脊柱炎(AS)滑膜组织中表达的蛋白,筛选RA滑膜中表达特异性下调的蛋白,并通过单核苷酸多态性(SNPs)分析方法分析靶蛋白编码基因对RA的遗传易感性.方法提取RA( 10例)、骨关节炎(10例)及AS(10例)滑膜组织总蛋白,每类疾病的样本等比例混合,然后用2-D凝胶电泳进行分析,对RA样本中表达量明显低于骨关节炎和AS的蛋白进行飞行时间质谱鉴定,检测结果用蛋白印迹法验证.用Taqman方法对中国人群中267例RA患者和160名健康对照的维生素D结合蛋白(VDBP)编码基因的标签SNPs进行基因分型,并扩大样本量(389例RA和371名健康对照)验证分型结果.采用单因素方差分析和Fisher确切概率法进行检验.结果蛋白质组学研究发现,在RA患者滑膜中VDBP表达量明显下降,蛋白印迹法也验证了这一结果.基因分型研究发现,SNP位点rs2282679与RA密切相关(P=0.026 794).进一步扩大样本实验也得出了一致结果,rs2282679与RA有相关性[比值比(OR )=0.678 639,95％可信区间(CI)0.541 113～0.851 118,P=0.000 776].结论与骨关节炎和AS患者相比,VDBP在RA患者滑膜中表达量明显下降.VDBP基因上的SNP位点rs2282679与RA有相关性.VDBP在RA中的表达下调及其基因的遗传效应显示VDBP可能参与RA的病理过程.
목적 본연구통과단백질조학적방법비교류풍습관절염(RA)、골관절염급강직성척주염(AS)활막조직중표체적단백,사선RA활막중표체특이성하조적단백,병통과단핵감산다태성(SNPs)분석방법분석파단백편마기인대RA적유전역감성.방법 제취RA( 10례)、골관절염(10례)급AS(10례)활막조직총단백,매류질병적양본등비례혼합,연후용2-D응효전영진행분석,대RA양본중표체량명현저우골관절염화AS적단백진행비행시간질보감정,검측결과용단백인적법험증.용Taqman방법대중국인군중267례RA환자화160명건강대조적유생소D결합단백(VDBP)편마기인적표첨SNPs진행기인분형,병확대양본량(389례RA화371명건강대조)험증분형결과.채용단인소방차분석화Fisher학절개솔법진행검험.결과 단백질조학연구발현,재RA환자활막중VDBP표체량명현하강,단백인적법야험증료저일결과.기인분형연구발현,SNP위점rs2282679여RA밀절상관(P=0.026 794).진일보확대양본실험야득출료일치결과,rs2282679여RA유상관성[비치비(OR )=0.678 639,95％가신구간(CI)0.541 113～0.851 118,P=0.000 776].결론 여골관절염화AS환자상비,VDBP재RA환자활막중표체량명현하강.VDBP기인상적SNP위점rs2282679여RA유상관성.VDBP재RA중적표체하조급기기인적유전효응현시VDBP가능삼여RA적병리과정.
Objective To screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles with that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients by a proteomic approach,and to explore the association of reduced expression with disease susceptibility by a single nucleotide polymorphism (SNP) analysis.Methods Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis.The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF-MS.The results were verified using Western blotting.Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA and 160 healthy controls.The genotyping results were confirmed in a large cohort of 389 patients with RA and 371 healthy controls.SPSS 11.5 software package was used for one way ANOVA and Fisher's exact test.Results The expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA was significantly decreased when examined by proteomic approach.This result was confirmed by Western blotting analysis.The rs2282679 was significantly associated with RA (P=0.026 794).This result was confirmed in a large cohort of RA( OR=0.678 639,95％CI 0.54l 113-0.851 118,P=0.000 776).Conclusion Compared with samples from patients with OA and AS,RA patients' synovial tissues have low VDBP expression when examined by the proteomic method.The tag SNP rs2282679 located in VDBP is significantly associated with RA.The decreased expression and the genetic effect of VDBP in RA suggest that a novel pathogenic pathway,in which vitamin D contributes,may be involved in the arthritis process of RA.