Morinobu Akio, Wang Biao, Liu Jun, Yoshiya Shinichi, Kurosaka Masahiro, Kumagai Shunichi
Department of Clinical Pathology and Immunology, Kobe University Graduate Schoool of Medicine, Kobe 650-0017, Japan.
J Rheumatol. 2006 Jun;33(6):1052-60.
To clarify the effects of trichostatin A (TSA), a histone deacetylase inhibitor, on the growth and survival of rheumatoid arthritis synovial fibroblasts (RA-SF).
Cell viability was assessed using a WST-8 assay and direct cell counting. Apoptosis was detected by annexin V staining on a flow cytometer. Protein and mRNA expression was determined by Western blotting, flow cytometry, and RT-PCR.
TSA suppressed cell growth of RA-SF in a dose-dependent manner, as determined by WST-8 assay and direct cell counting. Other histone deacetylase inhibitors also showed inhibitory effects on RA-SF proliferation. TSA upregulated p21(WAF1/CIP1) cell cycle inhibitor, suggesting that cell cycle arrest is involved in the reduction of cell numbers. In addition, TSA cooperated with Fas-induced pathway to induce cell death, determined by WST-8 assay and annexin V staining. TSA reduced FLICE inhibitory protein (FLIP) expression but not Bcl-2, Bcl-XL, and Fas expression, indicating that the synergistic effect may be through downregulation of FLIP.
TSA has antirheumatic effects on RA-SF and might be a potential therapeutic tool for the treatment of RA.
阐明组蛋白去乙酰化酶抑制剂曲古抑菌素A（TSA）对类风湿性关节炎滑膜成纤维细胞（RA-SF）生长和存活的影响。
使用WST-8检测法和直接细胞计数评估细胞活力。通过流式细胞仪上的膜联蛋白V染色检测细胞凋亡。通过蛋白质印迹法、流式细胞术和逆转录聚合酶链反应测定蛋白质和mRNA表达。
如通过WST-8检测法和直接细胞计数所确定的，TSA以剂量依赖性方式抑制RA-SF的细胞生长。其他组蛋白去乙酰化酶抑制剂也对RA-SF增殖显示出抑制作用。TSA上调p21（WAF1/CIP1）细胞周期抑制剂，表明细胞周期停滞参与细胞数量的减少。此外，如通过WST-8检测法和膜联蛋白V染色所确定的，TSA与Fas诱导的途径协同诱导细胞死亡。TSA降低FLICE抑制蛋白（FLIP）表达，但不降低Bcl-2、Bcl-XL和Fas表达，表明协同效应可能是通过下调FLIP实现的。
TSA对RA-SF具有抗风湿作用，可能是治疗类风湿性关节炎的一种潜在治疗工具。
