Histone deacetylase inhibitor suppression of auto antibody-mediated arthritis in mice via regulation of p16INK4a and p21WAF1/Cip1 expression
Histone deacetylase inhibitor suppression of auto antibody-mediated arthritis in mice via regulation of p16INK4a and p21WAF1/Cip1 expression
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DOI：
10.1002/art.20709
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发表时间：
2004-10-01
影响因子：
--
通讯作者：
Asahara, H
中科院分区：
文献类型：
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作者：
Nishida, K;Komiyama, T;Asahara, H
Objective.研究组蛋白脱乙酰酶（HDA）抑制剂缩肽（FK 228）是否对类风湿性关节炎（RA）患者滑膜成纤维细胞的增殖具有抑制作用，并研究FK 228全身给药在关节炎动物模型中的作用。在19只雄性DBA/1小鼠（6-7周龄）中诱导抗抗体介导的关节炎（AMA）;其中10只通过静脉注射FK 228（2.5 mg/kg）进行治疗，9只用作对照。FK 228的作用通过放射学、组织学和免疫组织化学分析以及关节炎评分进行检查。在关节置换手术时获得RA滑膜成纤维细胞（RASF）。MTT法检测FK 228对细胞增殖的影响。光镜和透射电镜观察细胞形态。通过实时荧光定量PCR和Western blot分析检测对细胞周期调节因子p16（INK 4a）和p21（WAF 1/Cip 1）表达的影响。染色质免疫沉淀法检测p16（INK 4a）和p21（WAF 1/Cip 1）启动子区的乙酰化状态。单次静脉注射FK 228（2.5 mg/ml）可成功抑制AMA小鼠的关节肿胀、滑膜炎症以及随后的骨和软骨破坏。FK 228处理诱导了AMA小鼠滑膜细胞中组蛋白的高度乙酰化，并降低了滑膜组织中肿瘤坏死因子α和白细胞介素-1 β的水平。FK 228对RASFs的体外增殖有抑制作用，且呈剂量依赖性。用FK 228处理细胞可诱导p16（INK 4a）的表达并上调p21（WAF 1/Cip 1）的表达。FK 228对p16（INK 4a）和p21（WAF 1/Cip 1）基因启动子区乙酰化的影响可能与FK 228对p16（INK 4a）和p21（WAF 1/Cip 1）基因启动子区乙酰化有关。我们的研究结果强烈表明，HDA抑制剂的全身给药可能代表一种新的治疗目标，通过诱导p16（INK 4a）表达和增加p21（WAF 1/Cip 1）表达，在RASF细胞周期阻滞的方式在RA。
Objective. To examine whether depsipeptide (FK228), a histone deacetylase (HDA) inhibitor, has inhibitory effects on the proliferation of synovial fibroblasts from rheumatoid arthritis (RA) patients, and to examine the effects of systemic administration of FK228 in an animal model of arthritis.Methods. Antoantibody-mediated arthritis (AMA) was induced in 19 male DBA/1 mice (6-7 weeks old); 10 of them were treated by intravenous administration of FK228 (2.5 mg/kg), and 9 were used as controls. The effects of FK228 were examined by radiographic, histologic, and immunohistochemical analyses and arthritis scores. RA synovial fibroblasts (RASFs) were obtained at the time of joint replacement surgery. In vitro effects of FK228 on cell proliferation were assessed by MTT assay. Cell morphology was examined by light and transmission electron microscopy. The effects on the expression of the cell cycle regulators p16(INK4a) and p21(WAF1/Cip1) were examined by real-time polymerase chain reaction and Western blot analysis. The acetylation status of the promoter regions of p16(INK4a) and p21(WAF1/Cip1) were determined by chromatin immunoprecipitation assay.Results. A single intravenous injection of FK228 (2.5 mg/ml) successfully inhibited joint swelling, synovial inflammation, and subsequent bone and cartilage destruction in mice with AMA. FK228 treatment induced histone hyperacetylation in the synovial cells and decreased the levels of tumor necrosis factor a and interleukin-1beta in the synovial tissues of mice with AMA. FK228 inhibited the in vitro proliferation of RASFs in a dose-dependent manner. Treatment of cells with FK228 induced the expression of p16(INK4a) and up-regulated the expression of p21(WAF1/Cip1). These effects of FK228 on p16(INK4a) and p21(WAF1/Cip1) were related to the acetylation of the promoter region of the genes.Conclusion. Our findings strongly suggest that systemic administration of HDA inhibitors may represent a novel therapeutic target in RA by means of cell cycle arrest in RASFs via induction of p16(INK4a) expression and increase in p21(WAF1/Cip1) expression.