丹酚酸B通过下调β-catenin抑制高糖诱导的内皮细胞间叶转化

    Salvianolic acid B inhibited high glucose-induced endothelial-mesenchymal transition through a down-regulation of β-catenin

      摘要
    • 摘要: 目的 模拟腹透液高糖环境，探索丹酚酸B在高糖诱导的内皮细胞间叶转化（endothelial-mesenchymal transition，EndMT）中的作用及β-catanin通路在其中可能的机制。方法 人脐静脉内皮细胞分3组：对照组（dulbecco modified eagle medium，DMEM），高糖组（含139 mmol/L葡萄糖DMEM）、高渗组（含等渗甘露醇DMEM），体外培养24 h、48 h后，蛋白质印迹法（western blot，WB）检测内皮细胞血小板-内皮细胞黏附分子（platelet endothelial cell adhesion molecule-1，CD31）、血管内皮钙黏蛋白（vascular endothelial cadherin，VE-cadherin）、α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、I型胶原蛋白（collagen I）及β-catenin蛋白表达。采用β-catenin特异性抑制剂XAV-939（1 μM）干预内皮细胞，WB再检测CD31、VE-cadherin、α-SMA、collagen I蛋白表达。另设丹酚酸B组（高糖+60 μg/mL丹酚酸B），细胞培养24 h、48 h后，WB检测CD31、VE-cadherin、α-SMA、collagen I及β-catenin蛋白表达。结果 在高糖环境下，内皮细胞CD31、VE-cadherin表达下降，α-SMA、collagen I表达上调。高糖可刺激内皮细胞β-catenin表达。选择性抑制β-catenin后，内皮细胞CD31、VE-cadherin的表达有所上调，α-SMA、collagen I表达下降。丹酚酸B下调高糖环境下内皮细胞β-catenin的表达，使高糖环境下内皮细胞CD31、VE-cadherin表达上调，α-SMA、collagen I表达下降。结论 高浓度葡萄糖上调β-catenin，诱导内皮细胞向间叶转化，丹酚酸B则可通过抑制β-catenin表达，在一定程度上改善高糖环境下内皮细胞转分化进程。

       

      Abstract: Objective To explore the role of salvianolic acid B during phenotypic transition of endothelial cells induced by high glucose and elucidate the possible mechanism of β-catenin pathway.Methods Human umbilical vein endothelial cells were divided into 3 groups of control (DMEM),high glucose (DMEM containing 139 mmol/L glucose) and high mannitol (DMEM containing isotonic mannitol).After culturing for 24/48h,protein expressions of CD31,VE-cadherin,α-SMA,collagen I and β-catenin in endothelial cells were detected by Western blot (WB).XAV-939 (1 μM),a specific inhibitor of β-catenin,was utilized for cellular interference.Also salvianolic acid B group (high glucose plus 60 μg/mL salvianolic acid B) was designated.Results In high glucose environment,the expressions of CD31 and VE-cadherin were down-regulated while those of α-SMA and collagen I up-regulated.High glucose stimulated the cellular expression of β-catenin.After a selective inhibition of β-catenin,the expressions of CD31 and VE-cadherin were up-regulated while the expressions of α-SMA and collagen I down-regulated.Salvianolic acid B down-regulated the expression of β-catenin,up-regulated the expressions of CD31 and VE-cadherin and lowered the expressions of α-SMA and collagen I under high glucose.Conclusion High glucose can up-regulate β-catenin and induce endothelial-mesenchymal transition in endothelial cells.And salvianolic acid B ameliorates phenotypic transition of endothelial cells through inhibiting the expression of β-catenin under high glucose.

       

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