摘要
目的:探究丁苯酞(N-butylphthalide,NBP)通过内质网应激途径对氧化低密度脂蛋白(ox-LDL)诱导内皮细胞凋亡的影响。方法:体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),给予NBP(5、10、20 mg/L)、4-苯丁酸(4-phenylbuturicacid,PBA,4 mmol/L)预处理1 h,再加入ox-LDL(100 mg/L)培养。CCK-8法检测细胞活力的变化,流式细胞术检测细胞凋亡,试剂盒检测培养基中乳酸脱氢酶(latic dehydrogenase,LDH)的水平,Hoechst 33342染色观察细胞凋亡的水平,Western blot法和RT-qPCR法检测内质网应激信号关键蛋白CHOP、GRP78的表达水平。结果:ox-LDL可以诱导HUVECs中内质网应激途径激活,从而促进细胞的凋亡;NBP呈剂量依赖性地抑制ox-LDL诱导的HUVECs损伤,细胞活力提高,凋亡率下调,同时LDH的水平降低;研究发现,NBP与PBA类似,可以抑制CHOP、GRP78的表达,抑制内质网应激的激活。结论:丁苯酞可以抵抗ox-LDL诱导的HUVECs凋亡,其机制与抑制内质网应激途径有关。
AIM : To investigate the effect of N-butylphthalide (NBP) on endothelial cell apoptosis induced by low density lipoprotein (ox-LDL) through endoplasmic reticulum stress pathway. METHODS : Human umbilical vein endothelial cells (HUVECs) were cultured in vitro , pretreated with NBP (5, 10, 20 mg/L) and 4-phenylbutyric acid (4-PBA, 4 mmol/L) for 1 hour, and then cultured with ox-LDL (100 mg/L). CCK-8 assay was used to detect cell viability, flow cytometry to detect apoptosis, LDH kit to detect the level of lactate dehydrogenase (LDH) in culture medium, Hoechst 33342 staining to observe the level of apoptosis. Western blot and RT-QPCR were used to detect the expression levels of CHOP and GRP78 in the endoplasmic reticulum stress signal. RESULTS :ox-LDL could induce the activation of ER stress pathway in HUVECs and promote the apoptosis of HUVECs. NBP inhibited the injury of HUVECs induced by ox-LDL in a dose-dependent manner, increased the cell viability, decreased the apoptosis rate and decreased the level of LDH. NBP, similar to PBA, could inhibit the expression of CHOP and GRP78 and the activation of ER stress. CONCLUSION : NBP can inhibit the apoptosis of HUVECs induced by ox-LDL, and the mechanism is related to the inhibition of ER stress pathway.
作者
官俏兵
杨毅
郭丽
韩晨阳
GUAN Qiaobing;YANG Yi;GUO Li;HAN Chenyang(The Second Hospital of Jiaxing, Jiaxing 314001, Zhejiang, China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2019年第4期397-402,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
浙江省科技厅实验动物项目(2017C37174)
浙江省卫生厅面上项目(2018KY804)
嘉兴市科技计划项目(2018AY32007)
(2017BY18023)
关键词
丁苯酞
氧化低密度脂蛋白
内质网应激
人脐静脉内皮细胞
N-butylphthalide
low density lipoprotein
endoplasmic reticulum stress
human umbilical vein endothelial cells
