Resveratrol combined with mesenchymal stem cells improves the metabolic memory effects on endothelial cell injury

doi:10.3969/j.issn.2095-4344.2014.14.016

Abstract

Abstract:

BACKGROUND: There are rare reports concerning whether resveratrol combined with mesenchymal stem cells is able to further improve the bad memory caused by high glucose-induced endothelial cell injury.
OBJECTIVE: To explore the effects of resveratrol combined with mesenchymal stem cells on high glucose-induced human umbilical vein endothelial cell injury.
METHODS: The human umbilical vein endothelial cells were divided into 10 groups: normal control glucose; mannitol control group; high glucose group; low-dose resveratrol group (0.1 μmol/L); middle-dose resveratrol group (1 μmol/L); high-dose resveratrol group (10 μmol/L); stem cells group; low-dose resveratrol+stem cells group; middle-dose resveratrol+stem cells group; high-dose resveratrol+stem cells group. Cell supernatants were collected to extract nuclear proteins on days 1, 4, 6 after culture in 5.5 mmol/L glucose. Western blot was used to investigate the expression of nuclear factor κB of human umbilical vein endothelial cells. Enzyme linked immunosorbent assay was used to detect the secretion of vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, plasminogen activator inhibitor-1.
RESULTS AND CONCLUSION: Compared with the normal control group, hyperglycemia memory up-regulated the expression of nuclear factor κB and increased the levels of vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, plasminogen activator inhibitor-1, which could still continue to rise after culturing in normal glucose. Resveratrol alone or combined with mesenchymal stem cells down-regulated the expression of
nuclear factor κB and decreased the levels of vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, plasminogen activator inhibitor-1 in a dose-dependent manner, which were lower than those in the high glucose group. There were no significant differences in nuclear factor κB, vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, plasminogen activator inhibitor-1 levels between stem cells group and high glucose group. Resveratrol can attenuate high glucose-induced injury to endothelial cells. The metabolic memory induced by high glucose may be mediated by nuclear factor κB pathway, which may be independent in the efficacy of mesenchymal stem cells in protecting endothelial cells.

Key words: stem cells, mesenchymal stem cells, endothelial cells, umbilical veins

CLC Number:

R394.2

Ding Hui, Wang Peng, Wang Yan-gang. Resveratrol combined with mesenchymal stem cells improves the metabolic memory effects on endothelial cell injury[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(14): 2232-2237.

Figures/Tables 2

2.1 细胞形态观察人脐静脉内皮细胞融合后呈多角形或短梭形，排列紧密，边界清晰，在倒置显微镜下呈单层铺路石样。选择生长良好的第3-5代人脐静脉内皮细胞用于实验。30 mmol/L葡萄糖刺激16 h后，细胞边界略肿胀，圆缩。经白藜芦醇预处理或联合间充质干细胞干预后，随着前者浓度增加，细胞形态得以改善。随着干预天数延长，可见细胞皱缩，并有死亡细胞及细胞碎片。 2.2 内皮细胞核因子κBp65表达情况正常组细胞核内核因子κBp65表达不明显，高糖组与正常组相比核因子κB p65表达明显增多(P < 0.05)，高剂量白藜芦醇及其联合干细胞组、甘露醇组与正常组相比差异无显著性意义。与高糖组相比，白藜芦醇及联合干细胞干预后内皮细胞核因子κB p65表达呈剂量依赖性降低(P < 0.05)，而干细胞组与高糖组相比核因子κB p65水平差异无显著性意义(图1)。 2.3 内皮细胞培养上清中血管细胞黏附分子1水平人脐静脉内皮细胞于高糖环境培养16 h后，血管细胞黏附分子1水平高于正常组(P < 0.05)，甘露醇组血管细胞黏附分子1水平与正常对照组相比差异无显著性意义。恢复正常糖浓度培养后，高糖组血管细胞黏附分子1水平持续上升，高于正常组(P < 0.05)，甘露醇组与正常对照组相比血管细胞黏附分子1水平差异无显著性意义。干细胞组与高糖组相比血管细胞黏附分子1水平差异无显著性意义，而用白藜芦醇或白藜芦醇联合干细胞干预后，与高糖组相比血管细胞黏附分子1水平大致呈白藜芦醇剂量依赖性降低(P < 0.05)，但低剂量白藜芦醇组及低剂量白藜芦醇+干细胞组与高糖组比较差异无显著性意义(表1)。 2.4 内皮细胞培养上清中单核细胞趋化蛋白1水平人脐静脉内皮细胞于高糖环境培养16 h后，单核细胞趋化蛋白1水平高于正常组(P < 0.05)，甘露醇组单核细胞趋化蛋白1水平与正常对照组相比差异无显著性意义。恢复正常糖浓度培养后，高糖组单核细胞趋化蛋白1水平持续上升，高于正常组P < 0.05)，甘露醇组与正常组相比单核细胞趋化蛋白1水平差异无显著性意义。低剂量白藜芦醇组在第4天以及中、高剂量白藜芦醇单用或联合干细胞组单核细胞趋化蛋白1水平低于高糖组(P < 0.05)，低剂量白藜芦醇组、干细胞组及两者联合组在第6天单核细胞趋化蛋白1水平低于高糖组，但差异无显著性意义(表1)。 2.5 内皮细胞培养上清中纤溶酶原激活剂抑制剂1水平与正常对照组相比，高糖培养16 h后，内皮细胞上清液中纤溶酶原激活剂抑制剂1水平升高(P < 0.05)，而甘露醇组纤溶酶原激活剂抑制剂1水平与正常对照组相比差异无显著性意义。恢复正常糖浓度培养后，高糖组纤溶酶原激活剂抑制剂1水平持续上升，且高于正常组(P < 0.05)，干细胞组和与正常组相比纤溶酶原激活剂抑制剂1水平差异有显著性意义 (P < 0.05)，甘露醇组与正常组相比纤溶酶原激活剂抑制剂1水平差异无显著性意义。低、中剂量白藜芦醇组及低剂量白藜芦醇+干细胞组在第4天纤溶酶原激活剂抑制剂1水平低于高糖组，但差异无显著性意义，低、中剂量白藜芦醇组"

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