基于NOD样受体3炎性小体通路对利拉鲁肽在氧化低密度脂蛋白诱导内皮细胞损伤的作用机制研究  

Mechanism of Liraglutide in Oxidized Low-density Lipoprotein Induced Endothelial Cell Injury Based on NOD-like Receptor 3 Inflammasome Pathway

作　　者：陈玲[1] 徐锐[1] 程新春[1] 张占英[1] 徐红[1] CHEN Ling;XU Rui;CHENG Xinchun;ZHANG Zhanying;XU Hong(Geriatric Medical Center,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院老年医学中心,新疆维吾尔自治区乌鲁木齐830001

出　　处：《中国全科医学》2025年第5期601-606,共6页Chinese General Practice

基　　金：新疆维吾尔自治区自然科学基金资助项目(2022D01C635);新疆维吾尔自治区人民医院院内基金项目(20200202)。

摘　　要：背景动脉粥样硬化是世界范围内引起心脑血管疾病最主要的原因,炎症是目前研究热点,其中NOD样受体3(NLRP3)是研究最为深入的炎症小体。胰高糖素样肽1(GLP-1)受体激动剂有抗动脉粥样硬化作用,具体机制尚不明确。目的研究利拉鲁肽通过拮抗氧化低密度脂蛋白(ox-LDL)诱导的内皮细胞损伤的作用机制。方法2022-03-25—05-19培养人脐静脉内皮细胞(HUVEC),取HUVEC加空白血清作为对照组,100μg/mL的ox-LDL干预HUVEC 48 h作为模型组,100μg/mL的ox-LDL干预HUVEC 24 h后分别加入100、200、400 nmol/L利拉鲁肽处理24 h作为利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组。CCK-8法计算细胞增殖率。通过扫描电镜观察焦亡细胞形态。检测乳酸脱氢酶(LDH)活力。酶联免疫吸附试验(ELISA)检测白介素(IL)-1β、IL-18表达水平。蛋白质免疫印迹试验(Western blot)检测NLRP3、接头蛋白凋亡相关斑点样蛋白(ASC)、天冬氨酸蛋白水解酶1(Caspase-1)、焦亡执行蛋白(GSDMD)、N端结构域的焦亡执行蛋白(N-GSDMD)表达水平。结果模型组、利拉鲁肽低浓度组和利拉鲁肽中浓度组细胞增殖率低于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组细胞增殖率高于模型组(P<0.05)。细胞扫描电镜结果示模型组细胞焦亡明显,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组细胞焦亡情况明显改善。模型组、利拉鲁肽低浓度组LDH活力高于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组低于模型组(P<0.05)。模型组、利拉鲁肽低浓度组IL-1β表达水平高于对照组,利拉鲁肽中浓度组、利拉鲁肽高浓度组IL-1β表达水平低于模型组(P<0.05);模型组IL-18表达水平高于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组IL-18表达水平低于模型组(P<0.05)。模型组NLRP3、ASC、Caspase-1Background Atherosclerosis is the primary cause of cardiovascular and cerebrovascular diseases worldwide,and inflammation is a current research focus,with NOD-like receptor 3(NLRP3)being the most intensively studied inflammasome.GLP-1 receptor agonists have shown anti-atherosclerotic effects,but the underlying mechanisms remain unclear.Objective To investigates the mechanism of liraglutide in antagonizing oxidized low-density lipoprotein(ox-LDL)induced endothelial cell injury.Methods From March 25 to May 19,2022,Human umbilical vein endothelial cells(HUVEC)were cultured,and HUVEC with blank serum was served as the control group,100μg/mL ox-LDL treated HUVEC for 48 hours was served as the model group.Liraglutide was added in concentrations of 100 nmol/L,200 nmol/L,and 400 nmol/L to the HUVECs treated with ox-LDL for 24 hours,forming low,medium,and high concentration liraglutide groups,respectively.Cell proliferation rates were calculated using the CCK-8 method.Pyroptotic cell morphology was observed by using scanning electron microscopy.Lactate dehydrogenase(LDH)activity was measured.The expression levels of interleukin(IL)-1βand IL-18 were detected using enzyme-linked immunosorbent assay(ELISA).Western blot was used to assess the expression levels of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),caspase-1,gasdermin D(GSDMD),and N-terminal GSDMD(N-GSDMD).Results Cell proliferation rate in the model group and both low and medium concentration liraglutide groups were lower than the control group,while the rate in all liraglutide-treated groups were higher than the model group(P<0.05).Scanning electron microscopy showed obvious pyroptosis in the model group cells,which was significantly reduced in all liraglutide-treated groups.LDH activity in the model group and the low concentration liraglutide group was higher than the control group,while it was lower in all liraglutide-treated groups compared to the model group(P<0.05).IL-1βlevel in the model group and the low concentration liraglutide g

关 键 词：动脉粥样硬化 利拉鲁肽 内皮细胞 氧化低密度脂蛋白 NOD样受体3 

分 类 号：R543.5[医药卫生—心血管疾病]