姜黄素在糖基化低密度脂蛋白诱导的血管平滑肌细胞脂质聚积中的作用机制研究  

Study on action mechanism of curcumin on lipid deposition of vascular smooth muscle cells induced by glycosylated low-density lipoprotein

作　　者：郑伟 朱明明 陈欣 柳丹 白磊 路可欣 李珊[2] 武福云[2] ZHENG Wei;ZHU Mingming;CHEN Xin;LIU Dan;BAI Lei;LU Kexin;LI Shan;WU Fuyun(Department of Integrated Medicine,Affiliated Dongfeng Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China;Institute of Basic Medical Sciences,Hubei University of Medicine,Shiyan,Hubei 442000,China;School of Medicine and Nursing,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院附属东风医院综合医疗科,湖北十堰442000 [2]湖北医药学院基础医学研究所,湖北十堰442000 [3]湖北医药学院药护学院,湖北十堰442000

出　　处：《重庆医学》2019年第3期377-381,共5页Chongqing medicine

基　　金：湖北省教育厅科学研究计划项目(D20142106);湖北医药学院(药护学院)大学生创新创业训练计划项目(201713249013);十堰市科技局项目(16Y59)

摘　　要：目的研究姜黄素在糖基化低密度脂蛋白(AGE-LDL)诱导的血管平滑肌细胞脂质聚积中的作用机制。方法体外制备并检测AGE-LDL,将AGE-LDL及不同浓度的姜黄素作用于大鼠血管平滑肌细胞系A7r5,分别设置空白对照组、AGE-LDL处理组、10μmol/L姜黄素处理组、20μmol/L姜黄素处理组和10、20μmol/L姜黄素+3-MA处理组;Western blot检测B类I型清道夫受体(SRB1)、糖原合成酶激酶3β(GSK3β)、磷酸化糖原合成酶激酶3β(p-GSK3β)、哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化哺乳动物雷帕霉素靶蛋白(pmTOR)、自噬相关蛋白Beclin和SQSTM1/p62蛋白表达水平;油红O染色检测脂质聚积程度。结果体外糖基化修饰的LDL硫代巴比妥反应显著降低、电泳迁移率明显增加,表明AGE-LDL构建成功。Western blot实验结果显示,与空白对照组相比,姜黄素处理组明显上调AGE-LDL诱导的SRB1的表达,并具有一定的浓度依赖效应。姜黄素还能够抑制p-GSK3β和p-mTOR的表达水平,增加细胞的自噬水平(beclin表达升高和p62表达下降)。油红O染色实验证实,AGE-LDL处理组脂质聚积明显,而姜黄素各处理组脂质聚积减弱,但3-MA各处理组脂质聚积进一步增加。结论姜黄素可促进血管平滑肌细胞SRB1表达,抑制其脂质聚积,机制可能与通过GSK-3β/mTOR途径增加自噬相关蛋白表达有关。Objective To investigate the mechanism of curcumin on glycosylated low density lipoprotein(AGE-LDL)induced lipid deposition in vascular smooth muscle cells.Methods AGE-LDL was prepared in vitro and measured.Then rat smooth muscle cell line A7r5 was treated by AGE-LDL and different concentrations of curcumin,and four groups were set up respectively,including the blank control group,AGE-LDL treatment group,10μmol/L curcumin treatment group,20μmol/L curcumin treatment group and 10,20μmol/L curcumin+3-MA treatment group.The protein expression levels of SRB1,GSK3β,p-GSK3β,mTOR,p-mTOR,beclin and p62 were detected by western blot.The oil red staining was used to detect the lipid deposition.Results In vitro AGE modified thiobarbital reaction was significantly reduced and electrophoretic mobility rate was significantly increased,indicating that AGE-LDL was successfully constructed.The western blot experiment analysis showed that compared with the blank control group,the curcumin treatment group obviously up-regulated the expression of SRB1 induced by AGE-LDL induced compared with the blank control group and had a concentration-dependent effect.Curcumin inhibited expression level of p-GSK3βand p-mTOR and enhanced the level of cellular autophagy(increased beclin expression and decreased p62 expression).The oil-red staining experiment verified that the lipid deposition in the AGE-LDL treatment group was obvious,while which in the various curcumin treatment groups was weakened,but the lipid deposition in the various 3-MA treatment group was further increased.Conclusion Curcumin could facilitate the expression of smooth muscle cell SRB1,inhibit its lipid deposition.Its mechanism might be related with increasing autophagy related protein expression through GSK-3β/mTOR pathway.

关 键 词：姜黄素 肌 平滑 血管 自噬 脂质聚集 

分 类 号：R543.3[医药卫生—心血管疾病]