白藜芦醇联合间充质干细胞改善损伤内皮细胞的代谢记忆效应

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

背景:白藜芦醇联合间充质干细胞是否能够进一步改善高糖诱导内皮细胞产生的不良记忆目前鲜有报道。目的:验证不同浓度的白藜芦醇联合间充质干细胞对高糖诱导人脐静脉内皮细胞损伤代谢记忆效应的保护作用。方法:将人脐静脉内皮细胞分为10组:正常对照组、甘露醇对照组、高糖组、白藜芦醇低剂量组(0.1μmol/L)、白藜芦醇中剂量组(1μmol/L)、白藜芦醇高剂量组(10μmol/L)、干细胞组、白藜芦醇低剂量+干细胞组、白藜芦醇中剂量+干细胞组及白藜芦醇高剂量+干细胞组。分别于5.5 mmol/L葡萄糖培养第1,4,6天收集细胞培养上清液、提取细胞核蛋白,应用ELISA法检测细胞培养上清中血管细胞黏附分子1、单核细胞趋化蛋白1、纤溶酶原激活剂抑制剂1的表达水平,以Western blot法检测细胞内核因子κB的蛋白水平。结果与结论:与正常对照组相比,高糖可诱导内皮细胞核因子κB表达上调,同时血管细胞黏附分子1、单核细胞趋化蛋白1和纤溶酶原激活剂抑制剂1水平升高,且在糖浓度恢复正常后仍持续上升。与高糖组相比,白藜芦醇及其联合干细胞干预后可以使内皮细胞核因子κB表达及血管细胞黏附分子1、单核细胞趋化蛋白1和纤溶酶原激活剂抑制剂1水平呈白藜芦醇剂量依赖性降低。干细胞组上述指标与高糖组比差异无显著性意义,甘露醇组与正常对照组相比差异无显著性意义。结果提示白藜芦醇可能通过核因子κB通路降低血管细胞黏附分子1、单核细胞趋化蛋白1和纤溶酶原激活剂抑制剂1水平,改善高糖代谢记忆效应介导的内皮细胞损伤。而间充质干细胞发挥内皮细胞保护作用可能不仅有赖于核因子κB通路。

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 pmol/L）; middle-dose resveratrol group （1 pmol/L）; high-dose resveratrol group （10 pmol/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 KB of human umbilical vein endothelial cells. Enzyme linked immunosorbent assay was used to detect the secretion of vascular cell adhesion molecule-I, monocyte chemoattractant protein-I, plasminogen activator inhibitor-l. RESULTS AND CONCLUSION： Compared with the normal control group, hyperglycemia memory up-regulated the expression of nuclear factor KB and increased the levels of vascular cell adhesion molecule-I, monocyte chemoattractant protein-I, plasminogen activator inhibitor-I, 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 KB and decreased the levels of vascular cell adhesion molecule-I, monocyte chemoattractant protein-I, 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 KB, vascular cell adhesion molecule-I, monocyte chemoattractant protein-I, 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 KB pathway, which may be independent in the efficacy of mesenchymal stem cells in protecting endothelial cells.