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目的 通过比较钙蛋白酶 (calpain)和钙蛋白酶抑制蛋白 (calpastatin)mRNA在类风湿关节炎 (RA)和骨关节炎 (OA)滑膜上的表达以及人的calpastatin抗原表位的探查 ,研究calpain和cal pastatin在RA发病机制中的作用。方法 总RNA从 3例RA和 2例OA患者滑膜组织中提取 ;RT PCR和数字影像光密度仪检测calpain和calpastatin的表达。用自动多肽合成仪合成人的calpastatinN端L区和C端Ⅳ区重叠的寡聚肽链 ;7例RA血清通过dot ELISA法鉴定合成的calpastatin肽链的抗原表位。结果 RA和OA患者的滑膜组织calpainmRNA的表达水平均高于calpastatin的表达 ,同时calpain的表达在RA高于OA。人的calpastatin的三个共同的抗原表位被鉴别在C 端Ⅳ区。 结论 Calpain和calpastatin系统参与了RA的发病 ;RA患者体内存在抗calpastatin自身抗体 ,合成人的calpastatin肽链可能为RA的诊治提供新的思路
Objective To investigate the expression of calpain and calpastatin mRNA in synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) as well as human calpastatin antigen epitopes by comparing calpain and calpastatin mRNA The role of cal pastatin in the pathogenesis of RA. Methods Total RNA was extracted from synovial tissue of 3 cases of RA and 2 cases of OA. The expression of calpain and calpastatin was detected by RT PCR and digital image densitometry. The overlapping oligopeptide chains of human calpastatin N and region C were synthesized by automatic peptide synthesizer. The epitope of calpastatin peptide was identified by dot ELISA in 7 RA sera. Results The expression of calpain mRNA in synovial tissue of patients with RA and OA was higher than that of calpastatin, while the expression of calpain was higher in RA than in OA. Three common epitopes of human calpastatin were identified at C-terminal region IV. Conclusions Calpain and calpastatin systems are involved in the pathogenesis of RA. Anti-calpastatin autoantibodies exist in RA patients and synthetic human calpastatin peptide chains may provide new ideas for the diagnosis and treatment of RA