烟酸姜黄素酯抑制LPS诱导的HUVECs分泌炎性细胞因子与内质网应激  

Nicotinate-curcumin inhibits LPS-induced inflammatory cytokines secretion and ER stress in HUVECs

作　　者：李招兵[1] 马小峰[1] 江振涛 黄云辉[1] LI Zhao-bing;MA Xiao-feng;JIANG Zhen-tao;HUANG Yun-hui(Department of Cardiology, the Affilited 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,湖南衡阳421001

出　　处：《基础医学与临床》2021年第1期62-67,共6页Basic and Clinical Medicine

基　　金：湖南省卫生计生委科研计划(B2016139)。

摘　　要：目的探讨烟酸姜黄素酯(NC)对LPS诱导的人脐静脉内皮细胞(HUVECs)分泌细胞因子的影响及其作用机制。方法体外培养HUVECs,用不同浓度NC孵育HUVECs 24 h。Western blot检测AMPK的磷酸化;用siRNA沉默磷酸腺苷活化蛋白激酶(AMPK)表达后,观察其对脂多糖(LPS)诱导NF-κB p65磷酸化、黏附分子表达、细胞培养上清中TNF-α和MCP-1分泌,以及内质网应激标志分子IRE-1、eIF2α和CHOP表达水平的影响。结果10~20μmol/L NC处理HUVECs 24 h后,AMPK的磷酸化水平显著增高(P<0.05),同时p65磷酸化有所减轻。沉默AMPK后,NC对NF-κB的抑制效应明显减弱(P<0.05)。同时,NC处理后,LPS所致ICAM-1,VCAM-1和E选择素等黏附分子表达明显降低(P<0.05),单核巨噬细胞THP-1与HUVECs的黏附作用也明显减轻,培养上清中TNF-α和MCP-1明显减少(P<0.05)。siRNA沉默AMPK后,上述效应有所减弱(P<0.05)。NC处理后可下调LPS所致的内质网应激标志分子IRE-1、eIF2α和CHOP的表达水平。而干扰AMPK表达后,可明显削弱NC对这些分子的抑制效应(P<0.05)。结论NC可抑制LPS诱导HUVECs表达和分泌黏附分子及细胞因子,并在一定程度上抑制内质网应激,其机制可能与激活AMPK有关。Objective To investigate the effect of nicotinate-curcumin(NC)on the LPS-induced secretion of cytokines by human umbilical vein endothelial cell(HUVECs)and its mechanism.Methods HUVECs were incubated with different concentrations of NC for 24 h.Western blot was used to detect the phosphorylation of AMPK.After silencing the expression of AMPK with siRNA,the potential outcomes on the phosphorylation of NF-kappa B p65,the expression of adhesion molecules,the secretion of TNF-alpha and MCP-1 in the supernatant of LPS-induced cells,and the expression of endoplasmic reticulum stress markers IRE-1,eIF2αand CHOP were observed.ResultsThe phosphorylation of AMPK was significantly increased and p65 phosphorylation was alleviated after HUVECs treatment with the concentration of 10~20 mmol/L NC for 24 h(P<0.05).After silencing of AMPK,the inhibition effectof NC on NF-κB was significantly reduced(P<0.05).At the same time,the expressions of ICAM-1,VCAM-1 and E-selectin,and the adhesion of THP-1 cells to HUVECs,content of TNF-αand MCP-1 in supernatants were significantly reduced after NC treatment(P<0.05).After siRNA silenced AMPK,the above effects were all inhibited(P<0.05).NC down-regulated the expression of endoplasmic reticulum stress markers IRE-1,eIF2αand CHOP induced by LPS.The inhibitory effect of curcumin on these molecules was significantly reduced by interfering with the expression of AMPK(P<0.05).Conclusions NC inhibits LPS-induced expression,secretion of adhesion molecules and cytokines in HUVECs,and also inhibits endoplasmic reticulum stress.The mechanism is potentially related to the activation of AMPK.

关 键 词：烟酸姜黄素酯 磷酸腺苷活化蛋白激酶 内质网应激 

分 类 号：R453.9[医药卫生—治疗学]