Trichostatin a prevents the accumulation of extracellular matrix in a mouse model of bleomycin-induced skin fibrosis
Trichostatin a prevents the accumulation of extracellular matrix in a mouse model of bleomycin-induced skin fibrosis
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DOI：
10.1002/art.22759
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发表时间：
2007-08-01
影响因子：
--
通讯作者：
Juengel, Astrid
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文献类型：
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作者：
Huber, Lars C.;Distler, Joerg H. W.;Juengel, Astrid
客观的。组织纤维化是许多疾病的标志性损害特征。在这项研究中，我们研究了组蛋白脱乙酰酶抑制剂曲古抑菌素 A (TSA) 在体外和体内对细胞因子驱动的纤维化反应的抗纤维化作用。方法。促纤维化细胞因子与 TSA 联合刺激系统性硬化症 (SSc) 患者和正常健康对照受试者的皮肤成纤维细胞。使用实时聚合酶链反应测量人 Col α 1(I) 和纤连蛋白，并使用 SirCol 胶原蛋白测定估计可溶性胶原蛋白的水平。使用电迁移率测定和共聚焦荧光显微镜研究 Smad 转录因子的细胞内分布。对于体内分析，通过对博来霉素诱导的纤维化模型中的小鼠皮肤组织进行组织学评估来量化皮肤纤维化。结果。添加 TSA 后，在正常和 SSc 皮肤成纤维细胞中观察到细胞因子诱导的 Col α 1(I) 和纤连蛋白转录减少。同样，TSA 刺激的 SSc 皮肤成纤维细胞中总胶原蛋白的表达降低至基础水平。 TSA 的作用机制包括抑制促纤维化 Smad 转录因子的核转位和 DNA 结合。蛋白质印迹分析显示 TSA 上调细胞周期抑制剂 p21，导致成纤维细胞增殖减少。此外，在博莱霉素诱导的小鼠纤维化中，TSA 可以阻止体内细胞外基质的真皮积累。结论。这些发现为纤维化的表观遗传调控提供了新的见解。 TSA 和类似的抑制化合物似乎代表了逆转细胞因子驱动的基质合成诱导（导致纤维化）的早期治疗策略。
Objective. Tissue fibrosis is a hallmark compromising feature of many disorders. In this study, we investigated the antifibrogenic effects of the histone deacetylase inhibitor trichostatin A (TSA) on cytokine-driven fibrotic responses in vitro and in vivo.Methods. Skin fibroblasts from patients with systemic sclerosis (SSc) and normal healthy control subjects were stimulated with profibrotic cytokines in combination with TSA. Human Col alpha 1(I) and fibronectin were measured using real-time polymerase chain reaction, and levels of soluble collagen were estimated using the SirCol collagen assay. Electromobilty shift assay and confocal fluorescence microscopy were used to investigate the intracellular distribution of Smad transcription factors. For in vivo analysis, skin fibrosis was quantified by histologic assessment of mouse skin tissue in a model of bleomycin-induced fibrosis.Results. Reductions in the cytokine-induced transcription of Col alpha 1(I) and fibronectin were observed in both normal and SSc skin fibroblasts following the addition of TSA. Similarly, the expression of total collagen protein in TSA-stimulated SSc skin fibroblasts was reduced to basal levels. The mechanism of action of TSA included inhibition of the nuclear translocation and DNA binding of profibrotic Smad transcription factors. Western blot analysis revealed an up-regulation of the cell cycle inhibitor p21 by TSA, leading to reduced proliferation of fibroblasts. In addition, in bleomycin-induced fibrosis in mice, TSA prevented dermal accumulation of extracellular matrix in vivo.Conclusion. These findings provide novel insights into the epigenetic regulation of fibrosis. TSA and similar inhibitory compounds appear to represent early therapeutic strategies for achieving reversal of the cytokine-driven induction of matrix synthesis that leads to fibrosis.