Lutty G A, Mathews M K, Merges C, McLeod D S
Wilmer Ophthalmological Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Curr Eye Res. 1998 Jun;17(6):594-607.
To evaluate the effects of adenosine and related substances on events that occur during vasculogenesis and angiogenesis, using in vitro assays.
Adenosine (ADO), inosine (INO, an adenosine catabolite), and 5'-(N-ethylcarboxamido) adenosine (NECA, an adenosine agonist) were evaluated for their effect on the proliferation of canine retinal microvascular endothelial cells (DRME), using a cell count assay. Also, these substances and ADO receptor selective agonists and antagonists were evaluated in an assay for DRME chemokinesis by measuring random migration into a wound made in a confluent cellular monolayer. Finally, the effects of these substances on DRME cord formation were evaluated in a 3-dimensional collagen gel. Bovine retinal extract (RE) was used as a positive control for all assays.
There was no effect on proliferation of DRME by any of the substances related to adenosine, but VEGF yielded a 30% stimulation of proliferation. Retinal extract, 10 microM ADO and 1.2 nM VEGF stimulation of DRME migration was 2- to 2.5-fold greater than 10 microM INO yielded. In addition, a combination of 1.2 nM VEGF with 10 microM ADO exceeded the stimulation in migration by ADO only and VEGF only. The total length of tubes formed in the presence of 10 microM ADO was comparable to that formed in the presence of RE and was 11-fold greater than with 10 microM INO. Tube length with a combination of VEGF plus ADO was 36% greater than with retinal extract. Use of selective ADO receptor antagonists suggested that tube formation and the migration response may be mediated through both adenosine A1 and A2 receptors, but use of selective ADO agonists suggests that A2 receptors may be more important than A1 for endothelial cell migration.
This in vitro analysis suggests that adenosine may stimulate retinal vasculogenesis, an event which involves migration of angioblasts and their assembly into vascular cords, prior to canalization.
通过体外试验评估腺苷及相关物质对血管发生和血管生成过程中所发生事件的影响。
使用细胞计数试验评估腺苷（ADO）、肌苷（INO，一种腺苷分解代谢产物）和5'-（N-乙基甲酰胺基）腺苷（NECA，一种腺苷激动剂）对犬视网膜微血管内皮细胞（DRME）增殖的影响。此外，通过测量随机迁移到汇合细胞单层中形成的伤口的情况，在DRME趋化试验中评估这些物质以及ADO受体选择性激动剂和拮抗剂。最后，在三维胶原凝胶中评估这些物质对DRME索形成的影响。牛视网膜提取物（RE）用作所有试验的阳性对照。
与腺苷相关的任何物质对DRME的增殖均无影响，但血管内皮生长因子（VEGF）可使增殖增加30%。视网膜提取物、10微摩尔/升的ADO和1.2纳摩尔/升的VEGF对DRME迁移的刺激作用比10微摩尔/升的INO产生的刺激作用大2至2.5倍。此外，1.2纳摩尔/升的VEGF与10微摩尔/升的ADO联合使用时，其对迁移的刺激作用超过了单独使用ADO和单独使用VEGF时的刺激作用。在存在10微摩尔/升ADO的情况下形成的管的总长度与存在RE时形成的总长度相当，并且比存在10微摩尔/升INO时形成的总长度大11倍。VEGF加ADO联合使用时的管长度比视网膜提取物时大36%。使用选择性ADO受体拮抗剂表明，管形成和迁移反应可能通过腺苷A1和A2受体介导，但使用选择性ADO激动剂表明，对于内皮细胞迁移，A2受体可能比A1受体更重要。
这种体外分析表明，腺苷可能刺激视网膜血管发生，这一过程涉及成血管细胞的迁移及其在管道形成之前组装成血管索。
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