Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China; National Local Joint Engineering Research Center for Precision Surgery & Regenerative Medicine, Xi'an, Shaanxi 710061, China; Shaanxi Province Center for Regenerative Medicine and Surgery Engineering Research, Xi'an, Shaanxi 710061, China.
Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China.
Life Sci. 2018 Oct 1;210:140-149. doi: 10.1016/j.lfs.2018.08.071. Epub 2018 Sep 1.
This study aims to explore the molecular mechanisms underlying sphingosine kinase 1 (SphK1) inducing pulmonary vascular remodeling and resveratrol suppressing pulmonary arterial hypertension (PAH).
monocrotaline (MCT) was used to induce PAH in rats. The right ventricular systolic pressure (RVSP), right ventricle hypertrophy index (RVHI) and histological analyses including hematoxylin and eosin staining, the percentage of medial wall thickness (%MT), α-SMA staining and Ki67 staining were performed to evaluate the development of PAH. Protein levels of SphK1, nuclear factor-kappaB (NF-κB)-p65 and cyclin D1 were determined using immunoblotting. Sphingosine-1-phosphate (S1P) concentration was measured using enzyme-linked immunosorbent assay.
SphK1 protein level, S1P production, NF-κB activation and cyclin D1 expression were significantly increased in MCT-induced PAH rats. Inhibition of SphK1 by PF543 suppressed S1P synthesis and NF-κB activation and down-regulated cyclin D1 expression in PAH rats. Suppression of NF-κB by pyrrolidine dithiocarbamate (PDTC) also reduced cyclin D1 expression in PAH model. Treatment of PAH rats with either PF543 or PDTC dramatically decreased RVSP, RVHI and %MT and reduced pulmonary arterial smooth muscle cells proliferation and pulmonary vessel muscularization. In addition, resveratrol effectively inhibited the development of PAH by suppression of SphK1/S1P-mediated NF-κB activation and subsequent cyclin D1 expression.
SphK1/S1P signaling induces the development of PAH by activation of NF-κB and subsequent up-regulation of cyclin D1 expression. Resveratrol inhibits the MCT-induced PAH by targeting on SphK1 and reverses the downstream changes of SphK1, indicating that resveratrol might be a therapeutic agent for the prevention of PAH.
本研究旨在探讨鞘氨醇激酶 1（SphK1）诱导肺血管重构和白藜芦醇抑制肺动脉高压（PAH）的分子机制。
使用野百合碱（MCT）诱导大鼠 PAH。通过右心室收缩压（RVSP）、右心室肥厚指数（RVHI）和组织学分析（包括苏木精和伊红染色、中膜厚度百分比（%MT）、α-SMA 染色和 Ki67 染色）评估 PAH 的发展。使用免疫印迹法测定 SphK1、核因子-κB（NF-κB）-p65 和细胞周期蛋白 D1 的蛋白水平。使用酶联免疫吸附试验测量鞘氨醇-1-磷酸（S1P）浓度。
MCT 诱导的 PAH 大鼠中 SphK1 蛋白水平、S1P 产生、NF-κB 激活和细胞周期蛋白 D1 表达显著增加。PF543 抑制 SphK1 抑制了 PAH 大鼠的 S1P 合成和 NF-κB 激活，并下调了细胞周期蛋白 D1 的表达。吡咯烷二硫代氨基甲酸盐（PDTC）抑制 NF-κB 也降低了 PAH 模型中的细胞周期蛋白 D1 表达。PAH 大鼠用 PF543 或 PDTC 治疗可显著降低 RVSP、RVHI 和 %MT，并减少肺动脉平滑肌细胞增殖和肺血管肌化。此外，白藜芦醇通过抑制 SphK1/S1P 介导的 NF-κB 激活和随后的细胞周期蛋白 D1 表达，有效抑制 PAH 的发展。
SphK1/S1P 信号通过激活 NF-κB 和随后上调细胞周期蛋白 D1 表达诱导 PAH 的发展。白藜芦醇通过靶向 SphK1 抑制 MCT 诱导的 PAH，并逆转 SphK1 的下游变化，表明白藜芦醇可能是预防 PAH 的治疗药物。
