Nitric oxide in vascular endothelial growth factor-induced focal angiogenesis and matrix metalloproteinase-9 activity in the mouse brain.
Nitric oxide in vascular endothelial growth factor-induced focal angiogenesis and matrix metalloproteinase-9 activity in the mouse brain.
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DOI：
10.1161/strokeaha.109.552059
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发表时间：
2009-08
期刊：
影响因子：
8.3
通讯作者：
Young WL
中科院分区：
文献类型：
--
作者：
Lee CZ;Xue Z;Hao Q;Yang GY;Young WL
血管内皮生长因子（VEGF）可诱导基质金属蛋白酶（MMP）-9活性和局灶性血管生成。我们假设VEGF激活脑MMP-9需要一氧化氮（NO）的参与。我们使用原位酶谱和内皮标记物CD 31比较了以下物质的体内作用：（1）NG-单甲基-L-精氨酸（L-NMMA），一种非特异性NO合酶（NOS）抑制剂;（2）L-N6-（1-亚氨基乙基）赖氨酸（L-NIL），一种诱导型NOS（iNOS）选择性抑制剂;和（3）强力霉素，一种已知的小鼠脑中MMP的非特异性抑制剂。3-硝基酪氨酸（3-NT）用作NO活性的替代物。炎性细胞标志物CD 68和MPO用于确认白细胞浸润。VEGF刺激的MMP-9活性主要在脑微血管周围表达。L-NMMA抑制脑血管生成（p<0.05），尤其是那些与VEGF诱导的MMP-9活化相关的微血管（p<0.02），与强力霉素的作用相当。L-NIL具有相似的抑制作用。3-NT证实了脑中的NO水平。与lacZ对照组相比，VEGF增加了诱导的脑血管生成病灶中的炎性细胞浸润，尤其是巨噬细胞。抑制NO的产生降低MMP-9的活性和VEGF刺激的脑中的局灶性血管生成。特异性和非特异性抑制NOS导致类似的减少，表明VEGF刺激的脑MMP活性和血管生成主要是通过iNOS介导的，iNOS是介导炎症反应的特异性NOS亚型。
Vascular endothelial growth factor (VEGF) can induce matrix metalloproteinase (MMP)-9 activities and focal angiogenesis. We hypothesized that VEGF activation of cerebral MMP-9 would require nitric oxide (NO) participation. We compared the in vivo effects of: (1) NG-monomethyl-L-arginine (L-NMMA), a non-specific NO synthase (NOS) inhibitor; (2) L-N6-(1-iminoethyl)lysine (L-NIL), an inducible NOS (iNOS) selective inhibitor; and (3) doxycycline, a known non-specific inhibitor of MMP in the mouse brain, using in situ zymography and endothelial marker CD31. 3-nitrotyrosine (3-NT) was used as a surrogate for NO activity. Inflammatory cell markers CD68 and MPO were used to confirm leukocyte infiltration. VEGF-stimulated MMP-9 activity expressed primarily around cerebral microvessels. L-NMMA suppressed cerebral angiogenesis (p<0.05), especially those microvessels associated with MMP-9 activation (p<0.02) induced by VEGF, comparable to the effect of doxycycline. L-NIL showed similar inhibitory effects. 3-NT confirmed NO levels in the brain. Compared to the lacZ control, VEGF increased inflammatory cell infiltration, especially macrophages, in the induced brain angiogenic focuses. Inhibition of NO production decreased MMP-9 activity and focal angiogenesis in the VEGF-stimulated brain. Both specific and non-specific inhibition of NOS resulted in similar reductions, suggesting that VEGF-stimulated cerebral MMP activity and angiogenesis are predominantly mediated through iNOS, a specific NOS isoform mediating inflammatory responses.