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

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(1μM)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+60μg/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。

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 24/48 h, 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.