扇贝糖胺聚糖对OX-LDL诱导血管内皮细胞氧化应激损伤的抑制作用机制研究

Study on Inhibitory Effects Mechanism of Scallop Skirt Glycosaminoglycan on Oxidative Stress Injury in Vein Endothelium Cells Induced by OX-LDL

目的:研究扇贝糖胺聚糖(SS-GAG)对人脐静脉血管内皮细胞(HUVEC)损伤的抑制作用机制。方法:试验分为阴性对照组、模型组与SS-GAG高、中、低浓度(质量浓度分别为200、100、50 mg/L)组,后3组以不同质量浓度SS-GAG培养细胞12 h后,以50μmol/L氧化低密度脂蛋白(OX-LDL)培养细胞24 h。以MTT法检测细胞活力;测定乳酸脱氢酶(LDH)活性;流式细胞仪测定活性氧(ROS)水平;实时荧光定量聚合酶链反应(RT-PCR)测定氧化低密度脂蛋白受体1(LOX-1)m RNA表达;Western blot测定NOX4蛋白表达。结果:与阴性对照组比较,模型组细胞活力减弱,LDH活性增强,ROS水平升高,LOX-1 m RNA表达增强,NOX4蛋白表达增强,差异有统计学意义(P<0.01)。与模型组比较,SS-GAG高、中、低浓度组细胞活力增强,LDH活性减弱,ROS水平降低,LOX-1 m RNA表达减弱,NOX4蛋白表达减弱,差异有统计学意义(P<0.01)。结论:SS-GAG有一定的保护血管内皮细胞作用,其机制可能与经LOX-1/NOX4途径抑制ROS产生、减轻氧化应激损伤有关。

OBJECTIVE：To investigate the inhibitory effects mechanism of scallop skirt glycosaminoglycan（SS-GAG）on inju-ry in human umbilical vein endothelium cells （HUVEC）. METHODS：In the test,there was a negative control group,a model group and the groups of SS-GAG at high,middle and low concentrations（mass concentrations of 200,100 and 50 mg/L respective-ly）. The cells in latter 3 groups were cultured in SS-GAG at different mass concentrations for 12 h,and then in 50 μmol/L oxidized low-density lipoprotein（OX-LDL）for 24 h. MTT method was used to detect cell viability and the activity of lactic dehydrogenase （LDH）,the flow cytometer to determine the level of reactive oxygen species （ROS）,real-time fluorescence quantitative poly-merase chain reaction （RT-PCR） to detect mRNA expression of lectin-like oxidized low-density lipoprotein receptor 1 （LOX-1）, and Western blot to detect NOX4 protein expression. RESULTS：Compared to the cells in the negative control group,those in the model group demonstrated lower viability,higher activity of LDH,higher level of ROS,and stronger expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance （P〈0.01）. Compared to the cells in the model group,those in the groups of SS-GAG at high,middle and low concentrations showed higher viability,lower activity of LDH,lower level of ROS and weaker expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance （P〈0.01）. CONCLUSIONS：SS-GAG can protect HUVEC to some degree by a mechanism which may be related to inhibiting ROS production via LOX-1/NOX4 pathway and relieving oxidative stress injury.