178 Angiotensin-(1-9) inhibits vascular smooth muscle cell proliferation and migration in vitro and neointimal formation in vivo
178 Angiotensin-(1-9) inhibits vascular smooth muscle cell proliferation and migration in vitro and neointimal formation in vivo
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178 血管紧张素-(1-9) 抑制体外血管平滑肌细胞增殖和迁移以及体内新内膜形成
DOI：
10.1136/heartjnl-2015-308066.178
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发表时间：
2015
期刊：
影响因子：
5.7
通讯作者：
McKinney C
中科院分区：
文献类型：
--
作者：
McKinney C
血管平滑肌细胞（SMC）的增殖和迁移是静脉移植失败和支架内再狭窄的发病机制的基础。目前的干预措施抑制SMC和内皮细胞（EC）的生长，导致血栓形成和再闭塞。血管紧张素II （AngII）是VSMC增殖和迁移的关键调节因子。肾素血管紧张素系统（RAS）的一个反调节轴已被确定，它抑制AngII，并以血管紧张素转换酶2和Ang-(1-7)为中心，作用于Mas。我们最近报道了Ang-(1-9)作为该轴的新成员，作用于血管紧张素2型受体（AT2R）以抑制心脏重构。本文研究了Ang-(1-9)在人SMC和EC体内迁移、增殖和血管损伤中的作用，并与Ang-(1-7)进行了比较。从人隐静脉中分离出SMC和EC。为了评估迁移，分别用Ang II和Ang-(1-9)或Ang-(1-7) +/- AT1R、AT2R或Mas拮抗剂氯沙坦、PD123,319 （PD）或A779刺激EC和SMC，并进行伤口愈合试验。为了评估增殖，用胎牛血清（FCS）和Ang-(1-9)或Ang-(1-7)±氯沙坦、PD或A779刺激EC和SMC。用MTS和Edu法测定细胞增殖。采用颈动脉钢丝损伤法建立动物模型。Ang-(1-7)或Ang-(1-9)±PD或A779通过微型泵皮下输送，并在损伤后28天量化新生内膜（NI）形成。Ang-(1-9)和Ang-(1-7)抑制Ang II诱导的VSMC迁移(Ang II 98.9±1.1%,Ang-(1-9) 43.3±3.3%,Ang-(1-7) 41.8±4.6%)；P < 0.001 vs Ang II)。两种多肽均能显著抑制FCS诱导的VSMC增殖（P < 0.05）。Ang-(1-9)和Ang-(1-7)对VSMC迁移和增殖的抑制作用分别被PD和A779选择性阻断，表明Ang-(1-9)通过AT2R起作用，Ang-(1-7)通过Mas起作用。Ang-(1-9)和Ang-(1-7)均不能阻止EC的迁移和增殖。小鼠颈动脉活体损伤28天诱导显著NI形成（NI/media area, NI/MA），损伤对照组（0.80±0.07）vs假手术组（0.01±0.01）；P < 0.001)；Ang-(1-9) （NI/MA 0.17±0.1，与损伤对照组相比P < 0.001）和Ang-(1-7) （NI/MA 0.40±0.07，与损伤对照组相比P < 0.05）减弱了这种毒性。PD可阻断Ang-(1-9)的作用（P < 0.001）， A779可阻断Ang-(1-7)的作用（P < 0.05），提示体内Ang-(1-9)通过AT2R起作用，Ang-(1-7)通过Mas起作用。我们证明了Ang-(1-9)在体外抑制VSMC增殖和迁移，并通过AT2R减少体内新内膜形成方面的一种新的直接作用。这些数据进一步揭示了Ang-(1-9)/AT2R相互作用在血管中的作用，并强调了Ang-(1-9)作为血管重构治疗剂的潜力。
Vascular smooth muscle cell (SMC) proliferation and migration underlie the pathogenesis of vein graft failure and in-stent restenosis. Current interventions inhibit both SMC and endothelial cell (EC) growth, leading to thrombosis and re-occlusion. Angiotensin II (AngII) is a key regulator of VSMC proliferation and migration. A counter-regulatory axis of the renin angiotensin system (RAS) has been identified which inhibits AngII and is centred around angiotensin converting enzyme2 and Ang-(1–7) acting at Mas. We recently reported Ang-(1–9) as a novel member of this axis, acting at the angiotensin type 2 receptor (AT2R) to inhibit cardiac remodelling. Here we investigated the role of Ang-(1–9) in human SMC and EC migration and proliferation and vascular injuryin vivo, and compared it to Ang-(1–7).SMC and EC were isolated from human saphenous veins. To assess migration, EC and SMC were stimulated with Ang II and Ang-(1–9) or Ang-(1–7) +/- the AT1R, AT2R or Mas antagonists losartan, PD123,319 (PD) or A779, respectively, and a wound healing assay performed. To assess proliferation, EC and SMC were stimulated with fetal calf serum (FCS) and Ang-(1–9) or Ang-(1–7) ± losartan, PD or A779. Proliferation was assessed using MTS and Edu assays. Anin vivomouse model was established via wire injury to the carotid artery. Ang-(1–7) or Ang-(1–9) ± PD or A779 were delivered subcutaneously via minipumps and neointima (NI) formation quantified 28 days post injury.Ang-(1–9) and Ang-(1–7) inhibited Ang II induced VSMC migration (Ang II 98.9 ± 1.1%, Ang-(1–9) 43.3 ± 3.3% and Ang-(1–7) 41.8 ± 4.6% wound closure; P < 0.001 vs Ang II). Furthermore, both peptides significantly inhibited FCS induced VSMC proliferation (P < 0.05). The inhibitory effects of Ang-(1–9) and Ang-(1–7) on VSMC migration and proliferation were selectively blocked by PD and A779, respectively, suggesting Ang-(1–9) acts via the AT2R and Ang-(1–7) via Mas. Neither Ang-(1–9) or Ang-(1–7) prevented EC migration or proliferation.In vivowire injury of the mouse carotid artery induced significant NI formation at 28days (NI/media area (NI/MA) 0.80 ± 0.07 injured control vs 0.01 ± 0.01 sham; P < 0.001); this was attenuated by Ang-(1–9) (NI/MA 0.17 ± 0.1; P < 0.001 vs injured control) and Ang-(1–7) (NI/MA 0.40 ± 0.07; P < 0.05 vs injured control). The effects of Ang-(1–9) were blocked by PD (P < 0.001 vs Ang-(1–9) alone) while the effects of Ang-(1–7) were blocked by A779 (P < 0.05 vs Ang-(1–7) alone), suggesting thatin vivoAng-(1–9) acts via the AT2R and Ang-(1–7) acts via Mas.We demonstrate for a novel, direct effect of Ang-(1–9) in inhibiting VSMC proliferation and migrationin vitro, and reducing neointimal formationin vivovia the AT2R. These data provide further insight into the role of the Ang-(1–9)/AT2R interaction in the vasculature and highlights the potential of Ang-(1–9) as a therapeutic agent for vascular remodelling.