Complementary effects of extracellular nucleotides and platelet-derived extracts on angiogenesis of vasa vasorum endothelial cells in vitro and subcutaneous Matrigel plugs in vivo.
Complementary effects of extracellular nucleotides and platelet-derived extracts on angiogenesis of vasa vasorum endothelial cells in vitro and subcutaneous Matrigel plugs in vivo.
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DOI：
10.1186/2045-824x-3-4
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发表时间：
2011-02-02
期刊：
影响因子：
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通讯作者：
Gerasimovskaya EV
中科院分区：
文献类型：
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作者：
Roedersheimer M;Nijmeh H;Burns N;Sidiakova AA;Stenmark KR;Gerasimovskaya EV
血小板通过释放多种生长因子、细胞因子和核苷酸（如ATP和ADP）来促进血管稳态和血管生成。最近的报道表明，总血小板提取物和选定的血小板生长因子对治疗性血管生成具有显著的促生长作用。然而，由于内源性腺嘌呤核苷酸在血小板分离和储存过程中迅速降解，我们研究了用外源性腺嘌呤核苷酸补充血小板衍生提取物是否会增加其促血管生成作用。采用肺动脉血管内皮细胞（VVEC），观察透析血小板源性可溶性提取物和细胞外腺嘌呤核苷酸对VVEC增殖、迁移和管腔形成的影响。此外，使用体内基质胶栓测定来检查血小板提取物和腺嘌呤核苷酸对皮下放置在50只ICR小鼠中的栓的新血管形成的影响。用组织学和统计学方法评价基质胶栓塞中血管结构的数量。血小板提取物（6.4-64 μg/ml）显著诱导DNA合成，并且在64 μg/ml的浓度下对VVEC增殖具有双相作用（48小时增加，随后在60小时减少）。用血小板提取物刺激VVEC也显著（高达几倍）增加了基质胶上的细胞迁移和管形成。用细胞外ATP（100 μM）刺激VVEC显著（高达10倍）增加了Matrigel上的迁移和管形成;然而，未观察到对细胞增殖的显著影响。我们还发现，ATP适度减少血小板提取物诱导的VVEC增殖（48小时）和迁移，但增强管形成。ATP或不可水解的核苷酸（ATPγS、ADPβS、MeSATP、MeSADP）的混合物均不诱导小鼠皮下注射的Matrigel塞的血管化，然而，这些核苷酸与血小板提取物的组合显著增加Matrigel塞中的功能性毛细血管的数量。这项研究的数据表明，血小板衍生的生长因子和细胞外核苷酸代表血管生成的重要调控信号。补充血小板提取物与外源性腺嘌呤核苷酸可能揭示新的可能性治疗血管生成和组织再生的方法。
Platelets contribute to vascular homeostasis and angiogenesis through the release of multiple growth factors, cytokines and nucleotides, such as ATP and ADP. Recent reports have demonstrated a marked growth-promoting effect of total platelet extracts and selected platelet growth factors on therapeutic angiogenesis. However, since endogenous adenine nucleotides are rapidly degraded during the platelet isolation and storage, we examined whether supplementing a platelet-derived extract with exogenous adenine nucleotides would augment their pro-angiogenic effects. Pulmonary artery vasa vasorum endothelial cells (VVEC) were used to examine the effects of dialyzed platelet-derived soluble extracts and extracellular adenine nucleotides on proliferation, migration and tube formation. In addition, an in vivo Matrigel plug assay was used to examine the effects of platelet extracts and adenine nucleotides on neovascularization of plugs subcutaneously placed in 50 ICR mice. The number of vascular structures in Matrigel plugs were evaluated by histological and statistical methods. Platelet extracts (6.4-64 μg/ml) significantly induced DNA synthesis and at a concentration of 64 μg/ml had a biphasic effect on VVEC proliferation (an increase at 48 hrs followed by a decrease at 60 hrs). Stimulation of VVEC with platelet extracts also significantly (up to several-fold) increased cell migration and tube formation on Matrigel. Stimulation of VVEC with extracellular ATP (100 μM) dramatically (up to ten-fold) increased migration and tube formation on Matrigel; however, no significant effects on cell proliferation were observed. We also found that ATP moderately diminished platelet extract-induced VVEC proliferation (48 hrs) and migration, but potentiated tube formation. Neither ATP, or a mixture of non-hydrolyzable nucleotides (ATPγS, ADPβS, MeSATP, MeSADP) induced vascularization of Matrigel plugs subcutaneously injected in mice, however, the combination of these nucleotides with platelet extracts dramatically increased the number of functional capillaries in the Matrigel plugs. Data from this study suggest that platelet-derived growth factors and extracellular nucleotides represent important regulatory signals for angiogenesis. Supplementation of platelet extracts with exogenous adenine nucleotides may reveal new possibilities for therapeutic angiogenesis and tissue regeneration approaches.