Mitomycin C induces pulmonary vascular endothelial-to-mesenchymal transition and pulmonary veno-occlusive disease via Smad3-dependent pathway in rats
Mitomycin C induces pulmonary vascular endothelial-to-mesenchymal transition and pulmonary veno-occlusive disease via Smad3-dependent pathway in rats
复制标题
丝裂霉素 C 通过 Smad3 依赖性途径诱导大鼠肺血管内皮间质转化和肺静脉闭塞性疾病
DOI：
10.1111/bph.15314
复制
发表时间：
2020-12-14
影响因子：
7.3
通讯作者：
Wang, Jian
中科院分区：
文献类型：
--
作者：
Zhang, Chenting;Lu, Wenju;Wang, Jian
背景与目的肺静脉闭塞性疾病（Pulmonary veno-occlusive disease，PVOD）是一种以肺小静脉阻塞导致肺动脉高压为特征的罕见疾病。实验方法采用丝裂霉素C（MMC）诱导的PVOD大鼠模型作为体内动物模型，以原代培养的大鼠肺微血管内皮细胞（PMVECs）为体外细胞模型，研究了肺微血管内皮细胞的内皮-间质转化（EndoMT），可能存在于从PVOD患者或MMC诱导的PVOD大鼠分离的肺微血管中。与对照血管相比，来自PVOD患者和PVOD大鼠的血管具有特异性内皮标志物血管性血友病因子（vWF）和间充质标志物α-平滑肌肌动蛋白（α-SMA）的共定位染色，表明存在共表达内皮和间充质标志物的细胞。在MMC诱导的PVOD大鼠和MMC处理的大鼠PMVEC的肺组织中，检测到内皮标志物（例如VE-钙粘蛋白和CD 31）水平降低和间充质标志物（例如波形蛋白、纤连蛋白和α-SMA）水平升高，表明存在EndoMT。此外，MMC诱导的TGF β/Smad 3/Snail轴的激活，同时用选择性Smad 3抑制剂（SIS 3）或针对Smad 3的小干扰RNA（siRNA）阻断该途径，显著地消除了MMC诱导的EndoMT。值得注意的是，治疗与SIS 3显着防止MMC诱导的PVOD在rats.Conclusions和影响的发病机制我们的数据表明，有针对性的抑制Smad 3导致一个潜在的，新的策略PVOD治疗，可能通过抑制肺微血管内皮MT。
Background and Purpose Pulmonary veno-occlusive disease (PVOD) is a rare disease characterized by the obstruction of small pulmonary veins leading to pulmonary hypertension. However, the mechanisms underlying pulmonary vessel occlusion remain largely unclear.Experimental Approach A mitomycin C (MMC)-induced PVOD rat model was used as in vivo animal model, and primarily cultured rat pulmonary microvascular endothelial cells (PMVECs) were used as in vitro cell model.Key Results Our data suggested an endothelial-to-mesenchymal transition (EndoMT) may be present in the pulmonary microvessels isolated from either PVOD patients or MMC-induced PVOD rats. In comparison to the control vessels, vessels from both PVOD patients and PVOD rats had co-localized staining of specific endothelial marker von Willebrand factor (vWF) and mesenchymal marker alpha-smooth muscle actin (alpha-SMA), suggesting the presence of cells that co-express endothelial and mesenchymal markers. In both the lung tissues of MMC-induced PVOD rats and MMC-treated rat PMVECs there were decreased levels of endothelial markers (e.g. VE-cadherin and CD31) and increased mesenchymal markers (e.g. vimentin, fibronectin and alpha-SMA) were detected indicating EndoMT. Moreover, MMC-induced activation of the TGF beta/Smad3/Snail axis, while blocking this pathway with either selective Smad3 inhibitor (SIS3) or small interfering RNA (siRNA) against Smad3, dramatically abolished the MMC-induced EndoMT. Notably, treatment with SIS3 remarkably prevented the pathogenesis of MMC-induced PVOD in rats.Conclusions and Implications Our data indicated that targeted inhibition of Smad3 leads to a potential, novel strategy for PVOD therapy, likely by inhibiting the EndoMT in pulmonary microvasculature.