COPD 患者诱导痰中细胞中组蛋白去乙酰化酶活性和诱导型一氧化氮合酶表达的改变与噻托溴铵治疗有关。

Altered histone deacetylase activity and iNOS expression in cells isolated from induced sputum of COPD patients treated with tiotropium.

机构信息

Department of Clinical Pharmacology, Medical University of Bialystok, 15a Waszyngtona St., Bialystok, 15-274, Poland,

出版信息

Adv Exp Med Biol. 2013;788:1-6. doi: 10.1007/978-94-007-6627-3_1.

Abstract

Chronic obstructive pulmonary disease (COPD) is the only major disease with increasing death rate. In COPD, progressive reduction in quality of life is closely related to the increasing limitation of airflow due to chronic bronchitis, cell hyperplasia, fibrosis, and irreversible lung damage. Signaling pathways involved in inflammatory processes in COPD and inflammatory response to therapy are unknown. Our aim was to isolate cells from induced sputum of COPD patients treated with formoterol or formoterol + tiotropium and assess enzymatic activity of histone deacetylases (HDACs) acetylated histone 4 (AcH4) and expression of inducible nitric oxide synthase (iNOS). HDACs are important in signal transduction and inflammation. iNOS is generating nitric oxide (NO) relevant to blood pressure regulation, inflammation and infections. Thirty stable COPD patients (21 males and 9 females, mean age 67 years) receiving 12 μg b.i.d. formoterol were assayed before and after 3 months add-on therapy consisting of 18 μg q.i.d. tiotropium. In all patients, spirometry, lung volumes, and DLCO were performed before and after tiotropium therapy and all patients were subjected to sputum induction. Sputum cells were isolated and processed to obtain cytosolic and nuclear fractions. HDAC activity was measured in nuclear fraction using colorimetric assay. Expression AcH4 and iNOS was quantified using Western blot. In patients receiving both drugs, FEV1 and lung volumes significantly improved compared with formoterol-only treated patients. Mean HDAC activity was slightly decreased (P < 0.05), while AcH4 levels and iNOS expression were significantly elevated in tiotropium-treated patients (increase by about 65 %; P < 0.01 and 77 %; P < 0.01 respectively). Our data show that beneficial effects of tiotropium in add-on therapy to formoterol may be related to altered histone signaling and increased iNOS expression.

摘要

慢性阻塞性肺疾病（COPD）是唯一一种死亡率不断上升的主要疾病。在 COPD 中，由于慢性支气管炎、细胞增生、纤维化和不可逆转的肺损伤，生活质量的逐渐下降与气流受限的不断增加密切相关。COPD 中炎症过程涉及的信号通路以及对治疗的炎症反应尚不清楚。我们的目的是从接受福莫特罗或福莫特罗加噻托溴铵治疗的 COPD 患者诱导痰中分离细胞，并评估组蛋白去乙酰化酶（HDACs）乙酰化组蛋白 4（AcH4）的酶活性和诱导型一氧化氮合酶（iNOS）的表达。HDACs 在信号转导和炎症中起着重要作用。iNOS 产生与血压调节、炎症和感染有关的一氧化氮（NO）。30 名稳定的 COPD 患者（21 名男性和 9 名女性，平均年龄 67 岁）接受 12 μg 每日 2 次福莫特罗治疗，在接受 18 μg 每日 4 次噻托溴铵附加治疗 3 个月前后进行了检测。在所有患者中，在接受噻托溴铵治疗前后进行了肺量测定、肺容积和 DLCO，所有患者均接受了痰诱导。分离痰液细胞并进行处理以获得胞质和核部分。使用比色法在核部分测量 HDAC 活性。使用 Western blot 定量 AcH4 和 iNOS 的表达。在接受两种药物治疗的患者中，与仅接受福莫特罗治疗的患者相比，FEV1 和肺容积显著改善。平均 HDAC 活性略有降低（P<0.05），而噻托溴铵治疗患者的 AcH4 水平和 iNOS 表达显著升高（分别增加约 65%；P<0.01 和 77%；P<0.01）。我们的数据表明，噻托溴铵在福莫特罗附加治疗中的有益作用可能与改变的组蛋白信号和增加的 iNOS 表达有关。