Vasostatin-1上调脐静脉内皮细胞表达HO-1减轻软脂酸所致炎症反应  

Vasostatin-1 alleviates palmitic acid-induced inflammation by inducing HO-1 expression in umbilical vein endothelial cells

作　　者：李招兵[1] 马小峰[1] 江振涛 黄云辉[1] LI Zhaobing;MA Xiaofeng;JIANG Zhentao;HUANG Yunhui(Department of Cardiology,Affiliated Nanhua Hospital,University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属南华医院心内科

出　　处：《免疫学杂志》2019年第11期972-977,共6页Immunological Journal

基　　金：湖南省卫生计生委科研计划(B2016139)

摘　　要：目的研究vasostatin-1(VS-1)对软脂酸诱导调脐静脉内皮细胞所致炎症反应和氧化应激的影响,并探讨其潜在的机制。方法体外培养人脐静脉内皮细胞(HUVECs),用100 nmol/L VS-1预孵育2 h,Western blot检测HUVECs中血红素氧合酶-1(HO-1)的表达,酶学法测定其活性的变化。随后检测VS-1处理前后Nrf2的核转位情况,并采用siRNA沉默其表达观察其对HO-1表达的影响。在研究VS-1对软脂酸所致的炎症反应时,首先以VS-1孵育细胞2 h,随后加入100μmol/L软脂酸作用12 h。ELISA测定IL-6和TNF-α的分泌;荧光探针测定胞内活性氧类(ROS)的产生;DNA结合实验测定NF-κB活性的变化。结果100 nmol/L VS-1孵育HUVECs细胞6 h即可诱导HO-1的表达并上调其酶活性,并持续至18 h。同时,VS-1也能上调细胞核内Nrf2的水平并增加其DNA结合活性;采用Nrf2 siRNA沉默其表达后,HO-1的表达水平和酶活性显著下降;软脂酸处理细胞后,细胞核内p65水平显著增高,细胞培养上清中IL-6和TNF-α含量以及胞内ROS含量明显增多,NF-κB的DNA结合活性也明显增强。而经100 nmol/L VS-1孵育后,NF-κB的活性有所降低,IL-6、TNF-α和ROS含量均有所减少;采用HO-1抑制剂SnPP共同孵育细胞后,VS-1对细胞因子以及ROS的抑制效应有所减弱。结论VS-1上调脐静脉内皮细胞表达HO-1,从而减轻软脂酸所致炎症反应。To investigate the effects of vasostatin-1(VS-1)on inflammatory response and oxidative stress induced by palmitic acid in umbilical vein endothelial cells and explore its potential mechanism,the human umbilical vein endothelial cells(HUVECs)were cultured in vitro and incubated with 100 nmol/L VS-1 for 2 hours.The expression of heme oxygenase-1(HO-1)in HUVECs was detected by Western blotting,and the activity of HO-1 was determined by enzymologic method.The nuclear translocation of Nrf2 was detected before and after VS-1 treatment,and the expression of Nrf2 was silenced by siRNA to observe its effect on HO-1 expression.In order to study the inflammatory response of VS-1 to palmitic acid,cells were incubated with VS-1 for 2 hours and then treated with 100 micromol/L palmitic acid for 12 hours.The secretion of IL-6 and TNF-αwere determined by ELISA,and the production of reactive oxygen species(ROS)was determined by fluorescence probe.The activity of NF-κB was measured by DNA binding assay.Data showed that after incubation with 100 nmol/L VS-1 for 6 hours,the expression of HO-1 was induced and the activity of HO-1 was up-regulated,which lasted for 18 hours.At the same time,VS-1 could also up-regulate the level of Nrf2 in the nucleus and increase its DNA binding activity.After silencing the expression of Nrf2,the expression level and activity of HO-1 decreased significantly.After treatment with palmitic acid,the level of p65 in the nucleus increased significantly,the contents of IL-6 and TNF-αin the supernatant and ROS in the cell culture increased significantly,and the DNA binding activity of NF-κB also increased significantly.After incubation with 100 nmol/L VS-1,the activity of NF-κB decreased,the contents of IL-6,TNF-αand ROS decreased.The inhibition effect of VS-1 on cytokines and ROS decreased after co-incubation with SnPP,a HO-1 inhibitor.These results demonstrate that VS-1 can up-regulate the expression of HO-1 in umbilical vein endothelial cells,thus alleviating the inflammatory reaction induced by palmi

关 键 词：Vasostatin-1 软脂酸 血红素氧合酶-1 

分 类 号：R392.23[医药卫生—免疫学]