摘要
目的通过比较强直性脊柱炎(AS)病人抗肿瘤坏死因子(TNF)-α单克隆抗体药物(remicade)治疗前后的膝关节滑膜细胞的基因谱变化,找出缓解AS病情相关的主要基质金属蛋白酶(MMP)-3,并探讨MMP-3潜在调控的作用和在AS发病中的作用机制和意义。方法从AS病人在remicade治疗前和治疗3个月后的关节滑膜细胞的基因表达谱(用含1176基因的cDNA微阵列检测)结果中筛选的靶基因即差异表达最显著的MMP-3为特异性刺激因子,用反转录-聚合酶链反应(RT-PCR)方法检测它诱导健康志愿者外周血单个核细胞(PBMC)的炎症相关基因TNF-α、c-jun和c-fos的表达。结果比较AS病人治疗前后的关节滑膜细胞基因差异表达的结果,MMP-3治疗后下调最显著(P=0.0147)。与MMP-3聚类在一起的基因有111个,和MMP-3相关性最强的基因是金属蛋白酶抑制剂前体(TIMP)-1;MMP-3能显著诱导PBMC的TNF-α、c-jun和c-fos的表达。结论在remicade治疗AS过程中,下调MMP-3是最重要的环节之一;通过检测MMP-3在关节滑膜细胞组织中的表达水平可作为AS诊断和病情进展监控的参考指标;MMP-3下调,可能使TNF-TNFR2-JNK-AP-1信息通道的激活状态减弱,从而减少致炎因子的释放,调整异常的基质降解和血管形成等病理因素,缓解AS炎症、减少软骨和骨质破坏的作用。
Objective To compare the gene expression profiles of knee synovial tissues of ankylosing spondylitis (AS) patients before and after the anti-TNF-?琢 monoclonal antibody drug remicade treatment.And to screen out the genes MMP-3 which were the potential gene target related with the pathogenesis of AS,evaluate its possible action and significance.Methods Knee synovial tissues obtained from patients with active AS before and after 3 months of treatment with remicade microarray experiments was carried out with 1176 target gene filters using total RNA of those with AS.After stimulated by microarray positive matrix metalloproteinases (MMP)-3,measurement of the expression of TNF-α,c-jun and c-fos in PBMC which obtained from healthy volunteers was performed with semi-quantitative PT-PCR.Results After comparing the gene differential expression of active AS patients before and after treatment,MMP-3 was the most promising microarray candidate,which became significantly down-regulated after remicade treatmext(P=0.0147).According to the results of clusting of microarray genes expression,there were 111 genes clusted in the same group with MMP-3.Metalloproteinase inhibitor-1 precursor (TIMP-1) had the strongest correlation with MMP-3;TNF-α,c-jun and c-fos were indeed strongly expressed in PBMC of healthy volunteers after stimulated by MMP-3.Conclusion During treatment with remicade,the most remarkable response is down-regulation of MMP-3 in the knee synovial tissues of patients with AS.Measurement of the expression level of MMP-3 is a valuable index for diagnosis and following up of the disease status.These data confirm the effect of anti-TNF-?琢 on the immune system,that the active state of TNF-TNFR2-JNK-AP-1 signal pathway has been weakened by down-regulation of MMP-3 after remicade treatment,thus decreasing the release of proinflammatory cytokines.This would alleviate cartilage and bone destruction,and induce proliferation and development of pannus.
出处
《中国药物与临床》
CAS
2004年第4期270-276,共7页
Chinese Remedies & Clinics
基金
国家自然科学基金委国家杰出青年科研基金资助项(30325019
30025041)
国家教委博士点基金资助项目(2002558082)
广州市科委基金资助项目(2002E3-E4021)
