吡格列酮对糖基化低密度脂蛋白诱导血管内皮细胞损伤的保护作用  

Protective effect of pioglitazone on glycosylated low density lipoprotein-induced injury of vascular endothelial cells

作　　者：刘城言 孙宪昌 于凤秀 朱文君 张颖 司艳红 Liu Chengyan;Sun Xianchang;Yu Fengxiu;Zhu Wenjun;Zhang Ying;Si Yanhong(Clinical Medicine College,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan,250117,China;Basic Medical College,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan,250117,China;Public Health College,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan,250117,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)临床医学院,山东济南250117 [2]山东第一医科大学(山东省医学科学院)基础医学院,山东济南250117 [3]山东第一医科大学(山东省医学科学院)公共卫生学院,山东济南250117

出　　处：《山东第一医科大学（山东省医学科学院）学报》2021年第12期894-898,共5页Journal of Shandong First Medical University ＆ Shandong Academy of Medical Sciences

基　　金：国家级大学生创新创业训练计划(201810439087);国家自然科学基金(81600681);山东省重点研发计划(2019GSF108260)。

摘　　要：目的观察吡格列酮(pioglitazone,PIO)对糖基化低密度脂蛋白(glycosylated low density lipoprotein,glc-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的保护作用,为临床应用PIO防治糖尿病血管病变提供实验依据。方法培养HUVECs,随机分为5组:对照组,模型组及PIO低、中、高剂量(1、10、100 μmol/L)组。MTT法检测细胞存活率,试剂盒检测细胞培养液中乳酸脱氢酶(lactate dehydrogen ase,LDH)活性及细胞内丙二醛(malondialdehyde,MDA)含量和超氧化物歧化酶(superoxide dismutase,SOD)活性,流式细胞仪检测细胞内活性氧(ractive oxygen species,ROS)水平,荧光定量PCR技术分析HUVECs中细胞间粘附分子-1(intercellular adhesion molecule-1,ICAM-1)、血管细胞粘附分子-1(vascular cell adhesion molecule-1,VCAM-1)mRNA表达.结果100 mg/L glc-LDL处理细胞24 h导致了HUVECs明显损伤,而PIO干预逆转了这一过程。100 μmol/L PIO使细胞存活率提高25%;10 μmol/L和100 μmol/L PIO使培养基中LDH活性分别降低18%和29%;100μmol/LPIO使细胞内ROS降低29%,MDA含量降低25%,SOD活性升高41%。此外,10μmol/LPIO使内皮细胞中VCAM-1 mRNA水平下调21%,100 μmol/L PIO使内皮细胞中IGAM-1和VCAM-1 mRNA的表达分别下调34%和32%。结论PI0通过提高细胞存活率,降低细胞膜通透性,减轻氧化应激和炎症反应,从而发挥保护血管内皮作用。Objective:To observe protective effect of pioglitazone(PIO)on glycosylated low density lipoprotein(glc-LDL)-induced injury of human umbilical vein endothelial cells(HUVECs).Methods:HUVECs were cultured and randomly divided into 5 groups:control group,model group and low,medium and high dose-PIO(1,10,100μmol/L)groups.Cell survival rate was analyzed by MTT.Lactate dehydrogenase(LDH)activity in cell culture medium,intracellular malondialdehyde(MDA)content and superoxide dismutase(SOD)activity were detected by the corresponding kits.Intracellular ROS level was analyzed by flow cytometry.The mRNA expressions of intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in HUVECs were tested by real time PCR.Results:24 h exposure of glc-LDL(100 mg/L)resulted in obvious damage of HUVECs,however,PIO treatment reversed this event.100 μmol/L PIO intervention increased cell viability by 25%.10 μmol/L and 100 μmol/L PIO decreased LDH activity in the medium by 18%and 29%respectively.100 pumol/L PIO also reduced intracellular ROS level by 29%,lowered MDA content by 25%,and enhanced SOD activity by 41%.Furthermore,10μmol/L PIO treatment downregulated VCAM-1 mRNA level by 21%and 100μmol/L PIO downregulated ICAM-1 mRNA by 34%and VCAM-1 mRNA by 32%.Conclusion:PIO treatment can protect vascular endothelial cells from glc-LDL injury in vitro by improving cell viability,reducing cell membrane permeability,alleviating oxidative stress and inhabiting inflammatory reaction.

关 键 词：吡格列酮 糖基化 低密度脂蛋白 内皮细胞 

分 类 号：R363[医药卫生—病理学]