Extracellular SOD-derived H2O2 promotes VEGF signaling in caveolae/lipid rafts and post-ischemic angiogenesis in mice.
Extracellular SOD-derived H2O2 promotes VEGF signaling in caveolae/lipid rafts and post-ischemic angiogenesis in mice.
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DOI：
10.1371/journal.pone.0010189
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发表时间：
2010-04-21
期刊：
影响因子：
3.7
通讯作者：
Ushio-Fukai M
中科院分区：
文献类型：
--
作者：
Oshikawa J;Urao N;Kim HW;Kaplan N;Razvi M;McKinney R;Poole LB;Fukai T;Ushio-Fukai M
活性氧（ROS），特别是H2 O2，对于参与内皮细胞（EC）增殖和迁移的VEGF受体2（VEGFR 2）信号传导的完全激活是必需的。细胞外超氧化物歧化酶（ecSOD）是一种主要的分泌型胞外酶，催化超氧化物歧化为H2 O2，并通过肝素结合结构域（HBD）锚定在EC表面。缺乏ecSOD的小鼠显示出生后血管生成受损。然而，目前尚不清楚ecSOD衍生的H2 O2是否调节VEGF信号转导。在此我们发现，基因转移ecSOD，而不是ecSOD缺乏HBD（ecSOD-ΔHBD），增加H2 O2水平在小鼠收肌，并促进血管生成后后肢缺血。缺乏ecSOD的小鼠在非缺血和缺血肢体中显示H2 O2减少。在体外，ECs培养液中ecSOD的过表达，而不是ecSOD-ΔHBD，增强了VEGF诱导的VEGFR 2酪氨酸磷酸化（VEGFR 2-pY），这是由清除细胞外H2 O2的过氧化氢酶短期预处理所阻止的。无论是可扩散的外源性H2 O2（<500 µM），还是一氧化氮供体，对VEGF诱导的VEGFR 2-pY都没有影响。这些表明，ecSOD通过HBD与EC结合是局部产生细胞外H2 O2以调节VEGFR 2-pY所必需的。从机制上讲，VEGF诱导的VEGFR 2-pY在小窝/脂筏，但非脂筏，增强ecSOD，其定位在脂筏通过HBD。ROS的作用靶点之一是蛋白酪氨酸磷酸酶（PTPs）。ecSOD在小窝/脂筏中诱导PTP 1B和DEP 1的氧化和失活，而非非脂筏，这两种酶负调节VEGFR 2-pY。破坏小窝/脂筏，或PTP抑制剂原钒酸盐，或PTP 1B和DEP 1的siRNA增强VEGF诱导的VEGFR 2-pY，其阻止ecSOD诱导的作用。在功能上，ecSOD促进VEGF刺激的EC迁移和增殖。总之，细胞外H2 O2产生的ecSOD定位在小窝/脂筏通过HBD促进VEGFR 2信号通过氧化失活的PTP在这些微域。因此，ecSOD是血管生成依赖性心血管疾病的潜在治疗靶点。
Reactive oxygen species (ROS), in particular, H2O2, is essential for full activation of VEGF receptor2 (VEGFR2) signaling involved in endothelial cell (EC) proliferation and migration. Extracellular superoxide dismutase (ecSOD) is a major secreted extracellular enzyme that catalyzes the dismutation of superoxide to H2O2, and anchors to EC surface through heparin-binding domain (HBD). Mice lacking ecSOD show impaired postnatal angiogenesis. However, it is unknown whether ecSOD-derived H2O2 regulates VEGF signaling. Here we show that gene transfer of ecSOD, but not ecSOD lacking HBD (ecSOD-ΔHBD), increases H2O2 levels in adductor muscle of mice, and promotes angiogenesis after hindlimb ischemia. Mice lacking ecSOD show reduction of H2O2 in non-ischemic and ischemic limbs. In vitro, overexpression of ecSOD, but not ecSOD-ΔHBD, in cultured medium in ECs enhances VEGF-induced tyrosine phosphorylation of VEGFR2 (VEGFR2-pY), which is prevented by short-term pretreatment with catalase that scavenges extracellular H2O2. Either exogenous H2O2 (<500 µM), which is diffusible, or nitric oxide donor has no effect on VEGF-induced VEGFR2-pY. These suggest that ecSOD binding to ECs via HBD is required for localized generation of extracellular H2O2 to regulate VEGFR2-pY. Mechanistically, VEGF-induced VEGFR2-pY in caveolae/lipid rafts, but non-lipid rafts, is enhanced by ecSOD, which localizes at lipid rafts via HBD. One of the targets of ROS is protein tyrosine phosphatases (PTPs). ecSOD induces oxidation and inactivation of both PTP1B and DEP1, which negatively regulates VEGFR2-pY, in caveolae/lipid rafts, but not non-lipid rafts. Disruption of caveolae/lipid rafts, or PTPs inhibitor orthovanadate, or siRNAs for PTP1B and DEP1 enhances VEGF-induced VEGFR2-pY, which prevents ecSOD-induced effect. Functionally, ecSOD promotes VEGF-stimulated EC migration and proliferation. In summary, extracellular H2O2 generated by ecSOD localized at caveolae/lipid rafts via HBD promotes VEGFR2 signaling via oxidative inactivation of PTPs in these microdomains. Thus, ecSOD is a potential therapeutic target for angiogenesis-dependent cardiovascular diseases.
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影响因子：
5.5
作者：
Baljinnyam, Erdene;Iwatsubo, Kousaku;Ishikawa, Yoshihiro
通讯作者：
Ishikawa, Yoshihiro
DOI：
10.1152/ajpheart.01277.2005
发表时间：
2006-09-01
影响因子：
4.8
作者：
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Caramelo, Carlos
影响因子：
4.1
作者：
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通讯作者：
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影响因子：
4.8
作者：
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通讯作者：
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影响因子：
11.8
作者：
Jakobsson, Lars;Kreuger, Johan;Claesson-Welsh, Lena
通讯作者：
Claesson-Welsh, Lena