摘要
目的 通过比较抗肿瘤坏死因子 (TNF) α单克隆抗体Remicade治疗强直性脊柱炎(AS)病人前后的膝关节滑膜细胞的基因谱变化 ,筛选出Remicade效应相关的差异表达基因 ,并探讨其在AS发病机制中的意义。方法 用 5例活动期AS病人在Remicade治疗前和 3个月后 (每次 5mg/kg ,第 0、2、6、12周静脉注射 ,以后每 8周用 1次 )的膝关节滑膜细胞 ,提取总RNA ;微阵列基因检测用含 1176个基因的cDNA芯片进行 ;数据用GeneSpring 5 1软件进行基因差异表达和聚类分析。结果 5例AS病人在Remicade治疗前后滑膜细胞基因差异表达和变化趋势总体聚类分析结果显示基因谱相关性强。用K mean聚类分析 ,其中第 5组与炎症相关密切的基因主要包括细胞因子、金属蛋白酶、黏附分子、与凋亡和致癌有关基因、受体、信号转导有关基因等。再按基因功能聚类 ,其中在致癌基因组共有 10个基因。比较AS病人治疗前后的关节滑膜细胞基因表达差异有显著性的基因(P <0 0 5 )共 4 5个 ,其中下调基因 4 4个 ;治疗后下调的基因主要包括三组 :①Th1细胞因子干扰素(IFN) γ ;②黏附分子 (VCAM 1、ICAM 1,Integrinβ4 ) ;③金属蛋白酶家系成员 3、7、8、11(MMP 3、7、8、11)和金属蛋白酶抑制体 (TIMP) 1。多数致炎基因包括TNF α、白细胞介素 (IL)
Objective To compare the gene patterns of knee synovial tissue of ankylosing spondylitis (AS) before and after therapy with anti TNF α monoclonal antibody remicade and to evaluate the potential AS related discriminating genes and their meaning in pathogenesis of AS.Methods Synovial tissue obtained from 5 patients with active AS before and after 3 months treatment with remicade (5 mg·kg -1 ·time -1 ,Ⅳ at weeks 0,2,6 and 12 and then per 8 weeks respectively).Total RNA was extracted from the synovial tissue.Microarray experiments were carried out with 1 176 target gene filters using synovial tissue of those with AS.Difference expression and clustering analysis with GeneSpring 5 1 microarray analysis software were used to find interesting genes.Results Before and after preliminary study of treatment the with remicade,total gene expression difference and clustering in synovial tissue of AS showed a high relativity.According to K mean analysis in group of number 5 the henes related with inflammation included cytokine,matrix metalloproteinase,adhesion molecule,apoptosis and oncogenes,receptors,signal transcription,etc.Ten genes have been clustered to an oncogenes group.There were 45 genes,which were significantly changed ( P values<0 05) in gene expression before and after remicade.Amond them,44 genes were down regulation after remicade.These included 3 group of genes:Th1 cytokine IFN γ;adhesion molecule VCAM 1,ICAM 1 and integrin β4;matrix metalloproteinases 3,7,8,11 (MMP 3,7,8,11) and metalloproteinase inhibitor 1 precursor (TIMP 1).The proinflammatory cytokine such as TNF α,IL 1β,IL 2R and MCP 1 also showed down regulation after treatment.Conclusion Gene microarray technique is effective for screening associated genes during the treatment with remicade,it helps to understand the pathogenesis of AS,and find the new genes associated with inflammation.
出处
《中华风湿病学杂志》
CAS
CSCD
2003年第10期591-595,T001,共6页
Chinese Journal of Rheumatology
基金
国家自然科学基金委国家杰出青年科研基金资助项目 ( 3 0 0 2 5 0 41)
国家教委博士点基金资助项目 ( 2 0 0 2 5 5 80 82 )
广州市科委资助项目 ( 2 0 0 2E3 E40 2 1)
