类叶升麻苷调控CREG表达对oxLDL诱导的人脐静脉内皮细胞损伤的保护作用  

Protective Effect of Acteoside on oxLDL-induced Human Umbilical Vein Endothelial Cells Injury by Regulating CREG Expression

作　　者：张振丽 任启正 孟祥智 高纪林 Zhang Zhenli;Ren Qizheng;Meng Xiangzhi;Gao Jilin(Department of Internal Medicine-Cardiovascular,Qingzhou Hospital of Traditional Chinese Medicine of Shandong Province,Shandong Qingzhou 262500,China;Shandong College of Traditional Chinese Medicine;Department of Cardiology,Weifang Yidu Central Hospital;Department of Acupuncture and Moxibustion,Qingzhou Hospital of Traditional Chinese Medicine of Shandong Province)

机构地区：[1]青州市中医院心内科,山东青州262500 [2]山东中医药高等专科学校 [3]潍坊市益都中心医院心内二科 [4]青州市中医院针灸一科

出　　处：《中国药师》2020年第9期1725-1730,共6页China Pharmacist

摘　　要：目的:探讨类叶升麻苷对氧化型低密度脂蛋白(oxLDL)诱导的人脐静脉内皮细胞(HUVEC)损伤的影响及机制。方法:不同浓度(1,10,100μmol·L^-1)的类叶升麻苷作用于oxLDL诱导的HUVEC细胞,酶联免疫吸附法检测细胞中丙二醛(MDA)含量和超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GSH-Px)活力,流式细胞术检测细胞凋亡,蛋白印迹(Western blot)法检测B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)和E1A激活基因阻遏子(CREG)蛋白表达水平,实时荧光定量PCR(RT-qPCR)检测CREG mRNA表达水平。转染CREG过表达载体或CREG小干扰RNA至HUVEC细胞,构建CREG过表达或CREG表达抑制的HUVEC细胞。oxLDL诱导CREG过表达的HUVEC细胞,或类叶升麻苷作用于oxLDL诱导的CREG表达抑制的HUVEC细胞,上述相同方法检测MDA含量、SOD和GSH-Px活力、细胞凋亡率、Bcl-2和Bax蛋白表达。结果:ox-LDL诱导HUVEC细胞后,MDA含量、细胞凋亡率和Bax蛋白水平升高,SOD和GSH-Px活力、Bcl-2蛋白及CREG mRNA和蛋白水平降低(P<0.05)。类叶升麻苷或过表达CREG可降低oxLDL诱导的HUVEC细胞MDA含量、凋亡率和Bax蛋白表达,提高SOD和GSH-Px活力及Bcl-2蛋白表达(P<0.05)。类叶升麻苷可促进oxLDL诱导的HUVEC细胞CREG mRNA和蛋白表达(P<0.05),抑制CREG表达降低了类叶升麻苷对oxLDL诱导的HUVEC细胞MDA含量、SOD和GSH-Px活力、细胞凋亡率及Bax和Bcl-2蛋白表达的影响(P<0.05)。结论:类叶升麻苷通过上调CREG表达降低oxLDL诱导的HUVEC细胞氧化应激和凋亡,减轻细胞损伤。Objective:To investigate the protective effect and mechanism of acteoside on oxLDL-induced HUVEC injury.Methods:Different concentrations of acteoside(1,10 and 100μmol·L-1)were applied to oxLDL-induced HUVEC.The content of MDA and activities of SOD and GSH in the cells were detected by enzyme-linked immunosorbent assay.The cells apoptosis was detected by flow cytometry.The expressions of Bcl-2,Bax and CREG protein were detected by Western blot.The expression of CREG Mrna was detected by RT-qPCR.CREG overexpression vector or CREG small interfering RNA was transfected into HUVEC cells to construct HUVEC cells with CREG overexpression or CREG expression inhibition.After oxLDL-induced HUVEC cells with CREG overexpression or acteoside were applied to oxLDL-induced HUVEC injury with CREG expression inhibition,the same methods as above were used to detect the content of MDA,activities of SOD and GSH,apoptosis rate and the expressions of Bcl-2 and Bax protein.Results:After ox-LDL induced HUVEC cells,MDA content,apoptosis rate and Bax protein level were increased,and SOD activity,SH-Px activity,Bcl-2 protein level,and the levels of CREG mRNA and protein were decreased(P<0.05).Acteoside or over-expressing CREG could reduce the MDA content,apoptosis rate and Bax protein expression in oxLDL-induced HUVEC cells,and increase the SOD activity,GSH-Px activity and Bcl-2 protein levels(P<0.05).Acteoside could promote the expressions of CREG mRNA and protein in oxLDLinduced HUVEC cells(P<0.05),and the inhibition of CREG expression reduced the effect of cycoside on the MDA content,SOD activity,GSH activity,apoptosis rate and expressions of Bax and Bcl-2 protein in oxLDL-induced HUVEC cells(P<0.05).Conclusion:Acteoside reduces the oxidative stress and the apoptosis of oxLDL-induced HUVEC cells by up-regulating CREG expression to reduce cell damage.

关 键 词：类叶升麻苷 E1A激活基因阻遏子 人脐静脉内皮细胞 氧化应激 细胞凋亡 

分 类 号：R965.1[医药卫生—药理学]