和厚朴酚对ox-LDL诱导的血管内皮细胞损伤的保护作用及其机制研究  

Protective effect of honokiol on vascular endothelial cell injury induced by oxidized low density lipoprotein and a study on mechanism

作　　者：朱为勇 唐元升[2] 盖延红 李越凡 尹刚 Zhu Weiyong;Tang Yuansheng;Gai Yanhong;Li Yuefan;Yin Gang(Department of Cardiovascular Medicine,Center Hospital of Qingdao City,Qingdao 266042,China)

机构地区：[1]青岛市中心医院心血管内科,青岛266042 [2]山东省立医院心血管内科,济南250021

出　　处：《中国循证心血管医学杂志》2018年第9期1079-1083,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：山东省科技发展计划项目(2016GSF118061)

摘　　要：目的探讨和厚朴酚对氧化低密度脂蛋白(ox-LDL)所诱导的人脐静脉血管内皮细胞(HUVECs)损伤的保护作用及其机制。方法体外培养HUVECs,采用细胞增殖-毒性检测(CCK8试剂盒法)检测不同浓度(0.1、1、10、50、100μmol/L)和厚朴酚对正常HUVECs的毒性作用;采用100 mg/L ox-LDL诱导24 h建立HUVECs损伤细胞模型,同时给予不同浓度和厚朴酚,CCK8法检测细胞增殖,流式细胞仪检测活性氧(ROS)水平,比色法测定细胞上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)以及谷胱甘肽(GSH)的水平;采用流式细胞仪检测凋亡水平;免疫印迹法检测凋亡相关蛋白和内质网应激(ERS)相应的信号通路改变。结果低浓度(0.1、1、10μmol/L)和厚朴酚对HUVECs细胞无毒性作用,高浓度(50、100μmol/L)和厚朴酚干预下细胞增殖能力减弱。与对照组比较,ox-LDL组细胞增殖受到明显抑制(P<0.05);而不同浓度和厚朴酚(0.1、1、10μmol/L)处理后,细胞增殖能力逐渐恢复(P<0.05)。Ox-LDL组较对照组ROS水平明显增加(P<0.05);而ROS水平随着厚朴酚浓度增加不断减少(P<0.05)。同时,与ox-LDL组比较,和厚朴酚处理后细胞上清中SOD、CAT和GSH水平增加,MDA水平降低(P均<0.05)。与ox-LDL组比较,和厚朴酚处理组HUVECs凋亡水平明显下降(P<0.05),Bcl-2/Bax比率增加,cleaved caspase3表达下降(P均<0.05)。而且与对照组比较,ox-LDL组内质网应激相关蛋白p-elF2α、p-PERK、ATF4和CHOP的表达增加,而和厚朴酚处理后上述蛋白表达均显著下降(P均<0.05)。结论和厚朴酚可能通过抑制氧化应激和内质网应激保护ox-LDL诱导的内皮细胞损伤。Objective To investigate the protective effect of honokiol on injury of human umbilical vein endothelial cells(HUVECs)induced by oxidized low density lipoprotein(ox-LDL)and study the mechanism.Methods HUVECs were culture in vitro,and cytotoxicity of honokiol,in difference doses(0.1μmol/L,1μmol/L,10μmol/L,50μmol/L and 100μmol/L)on HUVECs was detected by using cell counting kit-8(CCK-8).HUVECs were reduced by ox-LDL(100 mg/L)for 24 h for establishing HUVECs injury model,and then treated with honokiol in difference doses.HUVECs proliferation was detected by using CCK-8,level of reactive oxygen species(ROS)was detected by using flow cytometer,and levels of superoxide dismutase(SOD),malondialdehyde(MDA),catalase(CAT)and glutathione(GSH)were detected by using chromatoptometry.HUVECs apoptosis level was detected by using flow cytometer,and changes of apoptosis-related proteins and endoplasmic reticulum stress(ERS)-corresponded signal pathway were detected by using Western blotting assay.Results The low doses of honokiol(0.1μmol/L,1μmol/L and 10μmol/L)had no cytotoxic effect on HUVECs.HUVECs proliferation was reduced after intervention with high-dose of honokiol(50μmol/L and 100μmol/L).Compared with control group,HUVECs proliferation was significantly inhibited in ox-LDL group(P<0.05),and after treated with honokiol in different doses,HUVECs proliferation recovered gradually(P<0.05).The level of ROS increased significantly in ox-LD group compared with control group(P<0.05),and level of ROS decreased as the increase of honokiol doses(P<0.05).Meanwhile levels of SOD,CAT and GSH increased and MDA level decreased in all honokiol treatment groups compared with ox-LDL group(all P<0.05).HUVECs apoptosis level decreased significantly(P<0.05),Bcl-2/Bax increased and expression of cleaved caspase3 decreased(all P<0.05)in all honokiol treatment groups compared with ox-LDL group.The expressions of ERS-related proteins(p-elF2α,p-PERK,ATF4 and CHOP)increased in ox-LDL group compared with control group,and decreased signific

关 键 词：动脉粥样硬化 内皮细胞 和厚朴酚 氧化应激 内质网应激 

分 类 号：R543.5[医药卫生—心血管疾病]