Role of RASEF hypermethylation in cigarette smoke-induced pulmonary arterial smooth muscle remodeling
Role of RASEF hypermethylation in cigarette smoke-induced pulmonary arterial smooth muscle remodeling
复制标题
RASEF高甲基化在香烟烟雾诱导的肺动脉平滑肌重塑中的作用
DOI：
10.1186/s12931-019-1014-1
复制
发表时间：
2019-03-07
影响因子：
5.8
通讯作者：
Xie, Jungang
中科院分区：
文献类型：
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作者：
Li, Qinghai;Wu, Jixing;Xie, Jungang
背景肺动脉高压（pulmonary hypertension，PH）是一种进行性、致死性疾病。虽然香烟烟雾可以改变DNA甲基化状态，吸烟相关的PH这种分子改变的作用是uncleared.MethodsA PH大鼠模型暴露于香烟烟雾3个月。用右心导管测量右心室收缩压。评估暴露于香烟烟雾提取物的大鼠PA或原代人PA平滑肌细胞（HPASMCs）的组织学变化（右心室肥大指数、肺动脉（PA）中的中壁厚度）和DNMT 1蛋白水平。甲基化测序和MassArray®分别用于检测基因组和RASEF启动子甲基化状态。在DNMT 1敲除和香烟烟雾提取物暴露后，检查HPASMCs的行为（增殖，迁移）和RASEF甲基化状态;通过实时聚合酶链反应评估RASEF mRNA表达。RASEF过表达病毒载体被用来评估RASEF对大鼠PH和HPASMCs remodeling.ResultsHigher右心室收缩压，中壁厚度，右心室肥厚指数值吸烟组大鼠观察到的影响。烟雾暴露增加大鼠PA和HPASMCs中DNMT 1表达和RASEF甲基化水平。香烟烟雾提取物诱导HPASMCs行为改变和RASEF高甲基化，随后沉默，而DNMT 1敲低显著抑制这些变化。RASEF过表达明显抑制PH和HPASMCs重塑，可能通过磷酸化AKT（Ser 473），PCNA，和MMP 9 downregulation.ConclusionsCigarette smoke caused PA remodeling in PH rats related to RASEF hypermethylation.这些结果扩展了我们对香烟烟雾相关PH的关键表观遗传机制的理解，并可能为PH提供新的治疗靶点。
BackgroundPulmonary hypertension (PH) is a progressive and fatal disease. While cigarette smoke can change DNA methylation status, the role of such molecular alterations in smoke-associated PH is unclear.MethodsA PH rat model was developed by exposing animals to cigarette smoke for 3 months. Right ventricular systolic pressure was measured with a right heart catheter. Histological changes (right ventricular hypertrophy index, medial wall thickness in pulmonary arteries (PAs)) and DNMT1 protein levels in rat PAs or primary human PA smooth muscle cells (HPASMCs) exposed to cigarette smoke extract were assessed. Methylation sequencing and MassArray® were used to detect genomic and RASEF promoter methylation status, respectively. After DNMT1 knockdown and cigarette smoke extract exposure, HPASMCs behavior (proliferation, migration) and RASEF methylation status were examined; RASEF mRNA expression was evaluated by real-time-polymerase chain reaction. RASEF overexpression viral vectors were used to assess the impact of RASEF on rat PH and HPASMCs remodeling.ResultsHigher right ventricular systolic pressure, medial wall thickness, and right ventricular hypertrophy index values were observed in the smoking group rats. Smoke exposure increased DNMT1 expression and RASEF methylation levels in rat PAs and HPASMCs. Cigarette smoke extract induced HPASMCs behavioral changes and RASEF hypermethylation followed by silencing, while DNMT1 knockdown markedly inhibited these changes. RASEF overexpression distinctly inhibited PH and HPASMCs remodeling, possibly through phospho-AKT (Ser473), PCNA, and MMP9 downregulation.ConclusionsCigarette smoke caused PA remodeling in PH rats related to RASEF hypermethylation. These results expand our understanding of key epigenetic mechanisms in cigarette smoke-associated PH and potentially provide a novel therapeutic target for PH.