鸢尾素对脂多糖诱导的内皮细胞损伤影响  

Effects of Irisin on Lipopolysaccharide-Induced Endothelial Injury

作　　者：董文晟 高义鹏 叶云佳 李康 张鑫 DONG Wensheng;GAO Yipeng;YE Yunjia;LI Kang;ZHANG Xin(Department of Geriatrics,Renmin Hospital of Wuhan University,Hubei Key Laboratory of Metabolic and Chronic Diseases,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院老年病科,代谢与相关慢病湖北省重点实验室,湖北武汉430060

出　　处：《心血管病学进展》2025年第1期71-76,共6页Advances in Cardiovascular Diseases

基　　金：湖北省自然科学基金(2023AFB099);中央高校基本科研业务费专项资金(2042023kf0046);湖北省重点实验室开放项目(2023KFZZ028);武汉大学临床医学+青年托举项目。

摘　　要：目的探讨鸢尾素(irisin)对脂多糖(LPS)诱导的内皮细胞损伤影响。方法体外培养人脐静脉内皮细胞(HUVECs)并用LPS(100 ng/mL)处理12 h建立内皮细胞损伤模型,irisin干预组在刺激前给予irisin(20 nmol/L)预处理24 h。ELISA试剂盒检测细胞培养基中irisin含量;实时定量PCR检测各组炎症指标白细胞介素(IL)-1β、IL-6、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)和氧化应激指标NADPH氧化酶2(Nox2)、核转录因子红系2相关因子2(Nrf2)、超氧化物歧化酶2(SOD2)的mRNA水平,免疫印迹法检测FNDC5蛋白表达、p65蛋白磷酸化和核转位改变;试剂盒检测乳酸脱氢酶(LDH)漏出量、细胞丙二醛(MDA)含量以及过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性;DCFH-DA荧光探针检测活性氧(ROS)水平;CCK-8法检测HUVECs存活率。结果(1)LPS处理可抑制内皮细胞irisin合成分泌(P<0.05);(2)LPS组细胞存活率明显降低、LDH漏出量增加(P<0.05),而irisin处理组则明显改善(P<0.05);(3)irisin处理组细胞炎症指标IL-1β、MCP-1和TNF-α水平明显下调,p65蛋白磷酸化和核转位水平也受到显著抑制(P<0.05),但IL-6水平不受影响(P>0.05);(4)irisin处理不改变Nrf2 mRNA水平(P>0.05),但可降低Nox2表达并增加SOD2转录,irisin处理也明显降低细胞MDA含量并增加细胞抗氧化酶CAT和GSH-Px的活性,ROS生成也显著减少(P<0.05)。结论irisin对LPS诱导的内皮细胞炎症和氧化应激损伤具有明显的保护作用。Objective To investigate the effects of irisin on lipopolysaccharide(LPS)-induced endothelial injury.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and exposed to LPS(100 ng/mL)for 12 h to generate endothelial injury model.Cells with irisin intervention were pretreated with irisin(20 nmol/L)for 24 h.Irisin level in medium was measured by ELISA kit.The mRNA levels of inflammatory markers,including interleukin(IL)-1β,IL-6,monocyte chemotactic protein 1(MCP-1),tumor necrosis factor-α(TNF-α)and oxidative stress indicators,including reduced nicotinamide adenine dinucleotide phosphate oxidase 2(Nox2),nuclear factor-erythroid 2-related factor 2(Nrf2),superoxide dismutase 2(SOD2)were detected by real-time quantitative PCR,and Western blotting was used to assess FNDC5 protein expression as well as p65 phosphorylation and nuclear translocation.The amount of lactate dehydrogenase(LDH)leakage,intracellular malondialdehyde(MDA)content,and activities of catalase(CAT)and glutathione peroxidase(GSH-Px)were measured by commercial kits.DCFH-DA fluorescent probe was used to measure the intracellular reactive oxygen species(ROS)content.CCK-8 assay was used to detect the survival rate of HUVECs.Results(1)Irisin expression and secretion were significantly decreased by LPS insult(P<0.05).(2)LPS significantly decreased cell viability and increased LDH leakage(P<0.05),which was alleviated by irisin treatment(P<0.05).(3)Cells with irisin protection exhibited reduced levels of inflammatory markers,including IL-1β,MCP-1 and TNF-α,and p65 phosphorylation as well as nuclear translocation were also inhibited(P<0.05).However,IL-6 expression was not affected by irisin(P>0.05).(4)Irisin treatment had no effect on the mRNA level of Nrf2(P>0.05),but Nox2 expression was downregulated,and SOD2 transcription was upregulated in cells with irisin incubation.Besides,irisin treatment reduced intracellular MDA content and restored the activities of CAT together with GSH-Px.ROS generation was also significantly attenuated(

关 键 词：鸢尾素 脂多糖 内皮细胞 炎症 氧化应激 

分 类 号：R54[医药卫生—心血管疾病]