Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, South Australia, Australia.
School of Medicine, Faculty of Health and Medical Sciences, University of Adelaide, Adelaide, South Australia, Australia.
Cell Biol Int. 2021 Nov;45(11):2368-2379. doi: 10.1002/cbin.11682. Epub 2021 Aug 15.
Recently identified molecular targets in pulmonary artery hypertension (PAH) include sphingosine-1-phosphate (S1P) and zinc transporter ZIP12 signaling. This study sought to determine linkages between these pathways, and with BMPR2 signaling. Lung tissues from a rat model of monocrotaline-induced PAH and therapeutic treatment with bone marrow-derived endothelial-like progenitor cells transduced to overexpress BMPR2 were studied. Multifluorescence quantitative confocal microscopy (MQCM) was applied for analysis of protein expression and localization of markers of vascular remodeling (αSMA and BMPR2), parameters of zinc homeostasis (zinc transporter SLC39A/ZIP family members 1, 10, 12 and 14; and metallothionein MT3) and S1P extracellular signaling (SPHK1, SPNS2, S1P receptor isoforms 1, 2, 3, 5) in 20-200 µm pulmonary microvessels. ZIP12 expression in whole lung tissue lysates was assessed by western blot. Spearman nonparametric correlations between MQCM readouts and hemodynamic parameters, Fulton index (FI), and right ventricular systolic pressure (RVSP) were measured. In line with PAH status, pulmonary microvessels in monocrotaline-treated animals demonstrated significant (p < .05, n = 6 per group) upregulation of αSMA (twofold) and downregulation of BMPR2 (20%). Upregulated ZIP12 (92%), MT3 (57.7%), S1PR2 (54.8%), and S1PR3 (30.3%) were also observed. Significant positive and negative correlations were demonstrated between parameters of zinc homeostasis (ZIP12, MT3), S1P signaling (S1PRs, SPNS2), and vascular remodeling (αSMA, FI, RVSP). MQCM and western blot analysis showed that monocrotaline-induced ZIP12 upregulation could be partially negated by BMPR2-targeted therapy. Our results indicate that altered zinc transport/storage and S1P signaling in the monocrotaline-induced PAH rat model are linked to each other, and could be alleviated by BMPR2-targeted therapy.
最近在肺动脉高压（PAH）中确定的分子靶标包括鞘氨醇-1-磷酸（S1P）和锌转运蛋白 ZIP12 信号。本研究旨在确定这些途径之间的联系，以及与 BMPR2 信号之间的联系。研究了骨髓源性内皮样祖细胞转导以过表达 BMPR2 的大鼠模型中马兜铃酸诱导的 PAH 及其治疗治疗的肺组织。应用多荧光定量共焦显微镜（MQCM）分析血管重构标志物（αSMA 和 BMPR2）、锌稳态参数（锌转运蛋白 SLC39A/ZIP 家族成员 1、10、12 和 14；和金属硫蛋白 MT3）和 S1P 细胞外信号（SPHK1、SPNS2、S1P 受体亚型 1、2、3、5）在 20-200μm 肺微血管中的表达和定位。通过 Western blot 评估全肺组织裂解物中 ZIP12 的表达。测量 MQCM 读数与血流动力学参数（Fulton 指数（FI）和右心室收缩压（RVSP）之间的 Spearman 非参数相关性。与 PAH 状态一致，马兜铃酸处理的动物的肺微血管显示出明显的（p<0.05，每组 6 个）αSMA 上调（两倍）和 BMPR2 下调（20%）。还观察到上调的 ZIP12（92%）、MT3（57.7%）、S1PR2（54.8%）和 S1PR3（30.3%）。锌稳态参数（ZIP12、MT3）、S1P 信号（S1PRs、SPNS2）和血管重构（αSMA、FI、RVSP）之间存在显著的正相关和负相关。MQCM 和 Western blot 分析表明，BMPR2 靶向治疗可部分抵消马兜铃酸诱导的 ZIP12 上调。我们的结果表明，在马兜铃酸诱导的 PAH 大鼠模型中改变的锌转运/储存和 S1P 信号相互关联，并可通过 BMPR2 靶向治疗缓解。
