Tissue-type plasminogen activator contributes to remodeling of the rat ductus arteriosus.
Tissue-type plasminogen activator contributes to remodeling of the rat ductus arteriosus.
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DOI：
10.1371/journal.pone.0190871
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Ishikawa Y
中科院分区：
文献类型：
--
作者：
Saito J;Yokoyama U;Nicho N;Zheng YW;Ichikawa Y;Ito S;Umemura M;Fujita T;Ito S;Taniguchi H;Asou T;Masuda M;Ishikawa Y
动脉导管（DA）在出生后关闭以适应血液动力学的强烈变化，这需要内膜增厚（IT）的发生。已报道DA的平滑肌细胞在IT形成中起重要作用。然而，内皮细胞（ECs）的作用还没有得到充分的研究。我们在此集中于组织型纤溶酶原激活剂（t-PA），这是一个DA EC显性基因，并探讨其对IT形成的DA。EC从DA和主动脉分离胎鼠使用荧光激活细胞分选。RT-PCR结果显示，在足月大鼠胚胎（妊娠21天）的DA EC中t-PA mRNA的表达水平比主动脉EC高2.7倍。在早产和足月大鼠DA EC中检测到强的t-PA免疫反应。t-PA介导的纤溶酶原-纤溶酶转化激活明胶酶基质金属蛋白酶（MMP）。明胶酶谱显示，纤溶酶原补充显着促进大鼠DA EC的弹性蛋白溶解酶MMP-2的激活。原位酶谱分析表明，显着的明胶酶活性观察到在网站中断的内弹性板（IEL）在足月大鼠DA。在三维血管模型中，EC介导的纤溶酶原-纤溶酶转换增强IEL中断。在体内管理的纤溶酶原早产大鼠胎儿（妊娠第19天），其中IT是不好的形成，促进IEL破坏伴随着明胶酶激活和增强IT的形成在DA。此外，使用5个人类DA组织的实验表明，t-PA的表达水平是3.7倍，在IT领域比在图尼卡媒体。t-PA蛋白表达和明胶酶活性在人DA的IT区也被检测到。内皮细胞表达t-PA可能通过激活MMP-2和破坏IEL而促进DA的IT形成。
The ductus arteriosus (DA) closes after birth to adapt to the robust changes in hemodynamics, which require intimal thickening (IT) to occur. The smooth muscle cells of the DA have been reported to play important roles in IT formation. However, the roles of the endothelial cells (ECs) have not been fully investigated. We herein focused on tissue-type plasminogen activator (t-PA), which is a DA EC dominant gene, and investigated its contribution to IT formation in the DA. ECs from the DA and aorta were isolated from fetal rats using fluorescence-activated cell sorting. RT-PCR showed that the t-PA mRNA expression level was 2.7-fold higher in DA ECs than in aortic ECs from full-term rat fetuses (gestational day 21). A strong immunoreaction for t-PA was detected in pre-term and full-term rat DA ECs. t-PA-mediated plasminogen-plasmin conversion activates gelatinase matrix metalloproteinases (MMPs). Gelatin zymography revealed that plasminogen supplementation significantly promoted activation of the elastolytic enzyme MMP-2 in rat DA ECs. In situ zymography demonstrated that marked gelatinase activity was observed at the site of disruption in the internal elastic laminae (IEL) in full-term rat DA. In a three-dimensional vascular model, EC-mediated plasminogen-plasmin conversion augmented the IEL disruption. In vivo administration of plasminogen to pre-term rat fetuses (gestational day 19), in which IT is poorly formed, promoted IEL disruption accompanied by gelatinase activation and enhanced IT formation in the DA. Additionally, experiments using five human DA tissues demonstrated that the t-PA expression level was 3.7-fold higher in the IT area than in the tunica media. t-PA protein expression and gelatinase activity were also detected in the IT area of the human DAs. t-PA expressed in ECs may help to form IT of the DA via activation of MMP-2 and disruption of IEL.