A study of capillary pericyte viability on extracellular matrix produced by endothelial cells in high glucose
A study of capillary pericyte viability on extracellular matrix produced by endothelial cells in high glucose
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DOI：
10.1007/s00125-003-1043-6
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发表时间：
2003-03-01
期刊：
影响因子：
8.2
通讯作者：
Porta, M
中科院分区：
文献类型：
--
作者：
Beltramo, E;Buttiglieri, S;Porta, M
目的/假设。基底膜增厚和周细胞的选择性损失发生在糖尿病视网膜病变的早期。正如我们先前所示，周细胞粘附在高己糖浓度的内皮细胞产生的细胞外基质上受损，我们旨在验证粘附的改变是否会影响周细胞的活力和复制。通过在含有28 mmol/l D-葡萄糖（有或没有蛋白糖化抑制剂硫胺素或氨基胍）和高达28 mmol/l的D-半乳糖或L-葡萄糖的培养基中生长人脐静脉内皮细胞获得条件细胞外基质。去除内皮后，牛视网膜周细胞在这些基质上生长，在单独的实验中，在层粘连蛋白、纤连蛋白或IV型胶原上生长。分别于第7天、第2天、第18 h、第2天和第7天检测细胞计数和DNA合成情况，检测细胞周期穿越情况，检测细胞凋亡情况。在高D-葡萄糖和D-半乳糖中产生的基质上，周细胞计数和DNA合成减少，而在L-葡萄糖中获得的基质减少DNA合成，但不计数。硫胺素和氨基胍纠正降低周细胞活力时，加入到高D-葡萄糖。细胞周期和凋亡不受影响的周细胞生长在不同的条件矩阵。层粘连蛋白，纤连蛋白和IV型胶原蛋白没有修改周细胞replication.Conclusions/interpretation。减少周细胞计数可能取决于受损的初始粘附细胞外基质产生的内皮细胞在高浓度己糖，而不是受损的复制或活力。改变的细胞-基质相互作用可能促进糖尿病视网膜病变中的周细胞脱落，独立于高葡萄糖对周细胞复制的影响。
Aims/hypothesis. Thickening of the basement membrane and selective loss of pericytes occur early in diabetic retinopathy. As we showed previously that pericyte adhesion is impaired on extracellular matrix produced by endothelial cells in high hexose concentrations, we aimed to verify if altered adhesion could influence pericyte viability and replication.Methods. Conditioned extracellular matrices were obtained by growing human umbilical vein endothelial cells in media containing 28 mmol/l D-glucose, with or without the inhibitors of protein glycation thiamine or aminoguanidine, and D-galactose or L-glucose up to 28 mmol/l. Having removed the endothelium, bovine retinal pericytes were grown on these matrices and, in separate experiments, on laminin, fibronectin or type IV collagen. Pericyte viability and replication were measured by cell counts and DNA synthesis after 7 days, cell cycle traversal after 2 days and apoptosis after 18 h, 2 days and 7 days.Results. Pericyte counts and DNA synthesis were reduced on matrices produced in high D-glucose and D-galactose, whilst matrix obtained in L-glucose reduced DNA synthesis but not counts. Both thiamine and aminoguanidine corrected reduced pericyte viability when added to high D-glucose. Cell cycle and apoptosis were not affected by growing pericytes on different conditioned matrices. Laminin, fibronectin and type IV collagen did not modify pericyte replication.Conclusions/interpretations. Reduced pericyte counts could depend on impaired initial adhesion to the extracellular matrix produced by endothelium in high hexose concentrations, rather than impaired replication or viability. Altered cell-matrix interactions might facilitate pericyte dropout in diabetic retinopathy, independently of the effects of high glucose on pericyte replication.