青蒿琥酯对高糖条件下血管内皮细胞ICAM-1和MMP-9表达的影响

Effect of Artesunate on the Expression of ICAM-1 and MMP-9 in Vascular Endothelial Cells under High Glucose Condition

摘要: 目的：探讨ART对高糖条件下血管内皮细胞ICAM-1和MMP-9表达的影响。方法：将人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells, HUVEC)作为实验对象，分为葡萄糖(Glucose, G)组、40 mmol/L G + ART (G40 + ART)组，甘露醇(Mannitol, M)对照组、G40 + DMSO (DMSO)对照组，其中G组浓度梯度分为5.5 mmol/L G(G5.5)、25 mmol/L G(G25)、40 mmol/L G(G40)；M对照组浓度梯度为5.5 mmol/L G + 19.5 mmol/L M(M25)，5.5 mmol/L G + 34.5 mmol/L M(M40)，G40 + ART组中依据ART浓度梯度分为G40 + 10 ug/ml ART(10A)、G40 + 20 ug/ml ART(20A)、G40 + 40 ug/ml ART(40A)；DMSO对照组中DMSO用量与40A组中溶解ART所用DMSO体积相同。采用Western blot、细胞免疫荧光技术分别检测各组细胞间黏附分子-1(ICAM-1)、基质金属蛋白酶-9(MMP-9)蛋白表达情况。结果：1) ICAM-1、MMP-9蛋白在G25组较G5.5组表达升高(P < 0.01)，且G40组较G25组表达升高(P < 0.01)；ICAM-1、MMP-9蛋白在G40 + ART组较G40组表达下降，其中，10A组低于G40组(P < 0.01)，20A组低于10A组(P < 0.01)，40A组低于20A组(P < 0.01)。2) ICAM-1、MMP-9蛋白在G25组较M25组表达升高(P < 0.01)；G40组较M40组表达升高(P < 0.01)。3) DMSO对照组显示G40 + ART组ICAM-1、MMP-9蛋白表达低于DMSO组(P < 0.01)。结论：ICAM-1和MMP-9蛋白在高糖条件下表达升高，且具有浓度依赖性，ART可抑制高糖条件下血管内皮细胞ICAM-1和MMP-9蛋白的表达，且具有浓度依赖性，为进一步研究ART抑制视网膜新生血管形成和渗漏的机制中ICAM-1和MMP-9表达变化奠定基础。

Abstract: Objective: To investigate the effect of ART on the expression of ICAM-1 and MMP-9 in vascular en-dothelial cells under high glucose condition. Methods: Human Umbilical Vein Endothelial Cells (HUVEC) were divided into glucose (G) group, 40 mmol/L G + ART (G40 + ART) group, mannitol (M) control group, dimethyl sulfoxide (DMSO) control group. The concentration gradient of G group is 5.5 mmol/L G (G5.5), 25 mmol/L G (G25), 40 mmol/L G (G40); the concentration gradient of M control group is 5.5 mmol/L G + 19.5 mmol/L M (M25), 5.5 mmol/L G + 34.5 mmol/L M (M40); the concentration gradient of ART of G40 + ART group is G40 + 10 ug /ml ART (10A), G40 + 20 ug/ml ART (20A), G40 + 40 ug/ml ART (40A); the volume of DMSO in the DMSO control group is the same as it is in the 40A group. Western blot and cell immunofluorescence technique were used to detect the protein expression of Intercellular adhesion molecule-1 (ICAM-1) and Matrix metalloproteinase-9 (MMP-9) in each group. Results: The protein expression of ICAM-1 and MMP-9 in G25 group was higher than that in G5.5 group (P < 0.01), and it increased in G40 group compared with G25 group (P < 0.01); the protein expression of ICAM-1 and MMP-9 in G25 group was higher than that of M25 group (P < 0.01), and it increased in G40 group compared with M40 (P < 0.01); the protein expression of ICAM-1 and MMP-9 of G40 + ART group was lower than that of G40 group, in which it was lower in 20A group than that of 10A group (P < 0.01), and it was lower in 40A group compared with 20A group (P < 0.01). The DMSO control group showed that the protein expression of ICAM-1 and MMP-9 in G40 + ART was lower than that of G40 + DMSO group (P < 0.01). Conclusion: The expression of ICAM-1 and MMP-9 protein was increased under high glucose condition in a concentration-dependent manner. ART inhibited the expression of ICAM-1 and MMP-9 protein in vascular endothelial cells under high glucose condition in a concentration-dependent manner. This experiment lays the foundation for further study of the changes of ICAM-1 and MMP-9 expression in the mechanism of ART inhibiting retinal neovascularization and leakage.