白藜芦醇对脓毒症血管内皮屏障功能的保护作用及其机制研究  

Protective effect and related mechanism of resveratrol on vascular endothelial barrier dysfunction in sepsis

作　　者：郭智东 吴锦鸿 赵雪 吴泽生 王弋[1] Guo Zhidong;Wu Jinhong;Zhao Xue;Wu Zesheng;Wang Yi(Department of Emergency Medicine,Affiliated Hangzhou First People's Hospital,Zhejiang University School of Medicine,Hangzhou 310006,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院急诊科,杭州310006

出　　处：《中华危重症医学杂志（电子版）》2021年第4期281-289,共9页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：杭州市医药卫生科技计划项目(B20200683)。

摘　　要：目的探讨白藜芦醇在脂多糖诱导的血管内皮屏障功能损伤中的保护作用及其机制。方法将人脐静脉内皮细胞(HUVECs)分成对照组、脓毒症组及干预组。脓毒症组及干预组使用100μg/mL脂多糖刺激HUVECs制作脓毒症血管内皮细胞损伤模型,干预组在建模前给予白藜芦醇进行预处理。应用细胞计数试剂盒8(CCK-8)及5-乙炔基-2’脱氧尿嘧啶核苷(EdU)细胞增殖试剂盒检测细胞增殖能力,采用划痕实验及Transwell法检测细胞迁移能力,计算各组单核细胞对内皮细胞的黏附率,采用Western-blotting检测CyclinD1、含IQ模序的三磷酸鸟苷酶活化蛋白1(IQGAP1)、VE-cadherin、claudin-5、细胞间黏附分子1(ICAM-1)及血管细胞黏附分子1(VCAM-1)的蛋白表达水平,并比较各组细胞内活性氧的含量及核因子κB(NF-κB)入核率。结果CCK-8及EdU检测发现,各组细胞增殖能力的比较,差异均有统计学意义(F=92.460,P<0.001;F=40.910,P<0.001);划痕试验和Transwell法检测发现,各组细胞迁移能力的比较,差异均有统计学意义(F=27.050,P<0.001;F=56.000,P<0.001);且对照组及干预组的增殖能力及迁移能力均显著高于脓毒症组(P均<0.05)。各组细胞间单核细胞对内皮细胞的黏附率(F=19.240,P=0.035),CyclinD1(F=115.900,P<0.001)、IQGAP1(F=116.500,P<0.001)、VE-cadherin(F=108.500,P=0.042)、claudin-5(F=117.100,P=0.028)、ICAM-1(F=43.920,P<0.001)、VCAM-1(F=41.450,P=0.046)的蛋白表达水平,细胞内活性氧的含量(F=30.960,P=0.035)及NF-κB入核率(F=33.410,P=0.028)的比较,差异均有统计学意义。且进一步两两比较发现,与对照组及干预组比较,脓毒症组细胞的CyclinD1、IQGAP1、VE-cadherin及claudin-5蛋白表达水平均显著降低,而单核细胞对内皮细胞的黏附率,ICAM-1及VCAM-1蛋白表达水平,细胞内活性氧的含量及NF-κB入核率均显著升高(P均<0.05)。结论白藜芦醇通过抑制脂多糖诱导下活性氧聚集,减弱NF-κB激活及入核来保护脓毒症�Objective To investigate the protective effect and related mechanism of resveratrol on lipopolysaccharide-induced vascular endothelial barrier dysfunction in sepsis.Methods The human umbilical vein endothelial cells(HUVECs)were divided into control,sepsis and intervention groups.The cells in the sepsis group and intervention group were stimulated with 100μg/mL lipopolysaccharide to establish a vascular endothelial cell injury model of sepsis,and the cells in the intervention group were pretreated with resveratrol before modeling.The cell reproductive activity was detected by the cell counting kit-8(CCK-8)and 5-ethynyl-2’-deoxyuridine(Ed U),the cell migration ability was detected by the scratch test and Transwell chamber,and the adhesion rate of HUVECs to THP-1 cells was calculated in each group.The protein expression levels of Cyclin D1,IQ motif containing guanosine triphosphatase activating protein 1(IQGAP1),VE-cadherin,claudin-5,intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)were examined by Western-blotting,and the content of reactive oxygen species and the transfer rate of nuclear factor-kappa B(NF-κB)into the nucleus were compared among the three groups.Results The CCK-8 and Ed U showed that the cell reproductive activity among the three group was significantly different(F=92.460,P<0.001;F=40.910,P<0.001).The scratch test and Transwell chamber revealed that the cell migration ability among the three group was significantly different(F=27.050,P<0.001;F=56.000,P<0.001).The cell reproductive activity and cell migration ability in the control group and intervention group were higher than those in the sepsis group(all P<0.05).The adhesion rate of HUVECs to THP-1 cells(F=19.240,P=0.035),the protein expression of Cyclin D1(F=115.900,P<0.001),IQGAP1(F=116.500,P<0.001),VE-cadherin(F=108.500,P=0.042),claudin-5(F=117.100,P=0.028),ICAM-1(F=43.920,P<0.001)and VCAM-1(F=41.450,P=0.046),the content of reactive oxygen species(F=30.960,P=0.035)and the transfer rate of NF-κB into the

关 键 词：白藜芦醇 脓毒症 内皮 血管 

分 类 号：R459.7[医药卫生—急诊医学]