左归双降方对葡萄糖、胰岛素、低密度脂蛋白诱导人脐静脉内皮细胞损伤的干预作用

The Effects of Zuogui Shuangjiang Fang on the Vascular Endothelial Cells Injured by Glucose, Insulin and Low-Density Lipoprotein

目的 观察左归双降方提取液对葡萄糖(Glu)、胰岛素(Ins)、氧化低密度脂蛋白(ox-LDL)联合诱导下血管内皮细胞损伤的干预作用与作用机理。方法 以人脐带静脉内皮细胞株 ECV-304 为受试对象,以不同浓度的Glu、Ins、ox-LDL 联合作用 24 h,建立了一种糖尿病血管并发症的体外内皮细胞损伤模型,并在此基础上观测左归双降方提取液(ZGSJF)对该细胞损伤模型的调节作用。各试验组细胞继续培养 48 h 后分别检测细胞活性及其细胞培养液中的内皮素(ET-1)、一氧化氮(NO)以及细胞粘附因子(ICAM-1)的含量。结果 细胞继续培养 48h 后,模型组较正常组细胞活力明显下降(P<0.01),ET-1 及 ICAM-1 含量明显升高(P<0.01),NO 含量明显下降(P<0.01)。而不同剂量的 ZGSJF 给药后能显著改善细胞活力(与模型组比较,P<0.05),降低 ET、ICAM-1 及升高 NO 含量(与模型组比较,P<0.01),且显著优于对照药达美康。结论 左归双降方对血管内皮细胞具有一定的保护作用,而这一作用可能是通过促进内皮细胞分泌舒张性因子 NO、减少血管收缩因子 ET-1 的分泌、抑制与炎症反应相关的细胞粘附过程等途径,从而改善糖尿病合并高血压的血管损伤。

Objective To investigate the effects of Zuogui Shuangjiang Fang (ZGSJF) on vascular endothelial cells injured by glucose, insulin and low-density lipoprotein. Methods We made Human umbilical vein endothelial cell line ECV-304 exposed to different concentration of glucose, insulin and oxidized low-density lipoprotein (ox-LDL) for 24 hours, and to investigate the effects of ZGSJF on this injured vascular endothelial cells. The cells in each group were cultured for additional 48 hours. And then, cell viability was examined, and the supernatants were used to determine the contents of endothelin-1 (ET-1), nitric oxide (NO) and intercellular adhesion molecule-1 (ICAM-1) respectively. Results After the cells in each group were cultured for additional 48 hours, we found that the viability (measured by MTT test) of the cells in model group decreased markedly (P <0.01 vs blank group), and the secretion of ET-1 and ICAM-1 in the model group was significantly higher than that of the blank group (P <0.01), but the secretion of NO in the model group was significantly lower than that of the blank group (P <0.01). After treating of different dose ZGSJF, the cell viability was significantly greater than that of the model group (P <0.05), ET-1 and ICAM-1 were significantly lower than those of the model group (P <0.01), but NO were significantly higher than those of the model group (P <0.01). Conclusion The ZGSJF had the determinate protective function to the vascular endothelial cells, and this function might be realized by advancing NO and inhibiting ET and intercellular adhesion in order to decrease diabetic vasculopathy.