摘要
目的 研究类风湿性关节炎 (RA)患者关节滑膜成纤维细胞环氧化酶 2 (COX 2 )、诱导型一氧化氮合酶 (iNOS)及其诱导产物前列腺素E2 (PGE2 )、一氧化氮 (NO)的表达水平 ,探讨它们在RA致病过程中的作用。方法 分离滑膜成纤维细胞 ;用酶联免疫吸附法和酶还原法检测正常人关节滑膜液和滑膜细胞培养上清液中PGE2 、NO的含量 ;采用半定量逆转录 聚合酶联反应法检测滑膜组织及滑膜细胞COX 2、iNOSmRNA表达 ;并用细胞酶联免疫法检测细胞COX 2、iNOS蛋白表达水平。结果 RA患者关节滑膜组织COX 2、iNOSmRNA表达及滑膜液PGE2 、NO的水平分别为 0 42 ,0 83和(4 82± 1 74) μg/L、(80 4± 2 3 2 ) μmol/L ,明显高于正常对照组的 0 0 8,0 1 4和 (0 64± 0 1 3) μg/L、(2 6 2± 1 4 4) μmol/L(P <0 0 1 ) ;在炎性细胞因子刺激下RA患者滑膜成纤维细胞COX 2、iNOSmRNA水平 (0 48,0 98)、蛋白表达水平 (0 92± 0 1 8,0 78± 0 1 3)以及培养上清液中PGE2 、NO含量 [(2 1 44±4 2 6) μg/L ,(1 2 6 6± 32 2 ) μmol/L]均显著高于正常对照组 (P <0 0 1 )。 结论 滑膜成纤维细胞COX 2、iNOS的过度表达及PGE2 、NO的大量合成 。
Objectives To investigate the expression of cyclooxygenase 2 (COX 2), inducible nitric oxide synthase (iNOS) and their inducing products prostaglandins E 2 (PGE 2), nitric oxide (NO) in human rheumatoid synovial fibroblasts and their effects on the pathogenesis of rheumatoid arthritis (RA) Methods The productions of PGE 2, NO in the synovia and the culture supernatants of synovial fibroblasts isolated from RA patients and normal controls were measured with ELISA and enzyme reduction method The mRNA expressions of COX 2, iNOS in synovium and synovial fibroblasts were detected with semi quantitative reverse transcriptase polymerase chain reaction RT PCR And the COX 2, iNOS proteins were estimated with a cellular enzyme immunoassay using synovial fibroblasts as targets Results The COX 2, iNOS mRNA expressions in synovium(0 42,0 83)and PGE 2, NO productions in synovia(4 82±1 74)μg/L,(80 4±23 2) μmol/L from RA patients increased markedly when compared with those in normal controls 0 08,0 14 and (0 64±0 13) μg/L,(26 2±14 4)μmol/L,all( P <0 01 ) The mRNA and protein expressions of COX 2, iNOS in synovial fibroblast(0 48,0 98 and 0 92±0 18,0 78±0 13)and PGE 2, NO productions in cell culture supernatants(21 44±4 26)μg/L,(126 6±32 2)μmol/L also increased significantly when compared with those in normal controls( P <0 01) Conclusion The over expression of COX 2, iNOS and overproduction of PGE 2, NO may be an important factor in the pathogenetic process of RA
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2002年第6期345-348,共4页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金资助项目 ( 3980 0 135)