白藜芦醇靶向mTORC2/Rictor抑制HUVECs的增殖迁移及管腔形成  

Inhibitory effects of resveratrol on the proliferation,migration and angiogenesis of HUVECs through targeting mTORC2/Rictor

作　　者：李明珠 刘丽乔[2] 刘卓琦[2] 王群[3] LI Mingzhu;LIU Liqiao;LIU Zhuoqi;WANG Qun(First Clinical Medical College Nanchang University,Nanchang 330006,China;Dept.of Biochemistry&Molecular Biology,Basic Medical College Nanchang University,Nanchang 330006,China;Dept.of Cardiovascular Surgery,First Affiliated Hospital of Nanchang University,Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一临床医学院,江西南昌330006 [2]南昌大学基础医学院生物化学与分子生物学教研室,江西南昌330006 [3]南昌大学第一附属医院心胸外科,江西南昌330006

出　　处：《基础医学与临床》2018年第4期445-450,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81760078);江西省教育厅基金(700649003)

摘　　要：目的探讨白藜芦醇对腺病毒介导的Rictor基因转染人脐静脉内皮细胞(HUVECs)的增殖、迁移及管腔形成的影响。方法将PCR扩增得到的目的基因Rictor定向克隆入GV314载体获得重组质粒,经Ad Max包装系统与辅助包装质粒在HEK 293T细胞中重组构建Rictor过表达腺病毒载体(Ad-Rictor),不含目的基因的Ad-Null为阴性对照组。分离培养HUVECs后分别用Ad-Rictor及Ad-Null重组腺病毒感染细胞,另设空白对照组及白藜芦醇干预组Ad-Rictor+Res。感染后荧光显微镜及Western blot检测重组蛋白表达;CCK8、划痕实验和血管形成实验观察HUVECs增殖、迁移及血管形成能力。结果重组腺病毒Ad-Rictor及Ad-Null构建成功。与对照组相比,Ad-Rictor感染HUVECs显著上调基因Rictor的表达,提高了HUVECs的增殖活力,迁移和体外管腔形成能力(P<0.05);白藜芦醇干预显著抑制了Rictor过表达情况下HUVECs的增殖、迁移和体外管腔形成(P<0.05)。结论白藜芦醇可以靶向mTORC2/Rictor抑制人脐静脉内皮细胞增殖、迁移及管腔形成。Objective To investigate the effect of resveratrol on the proliferation,migration and angiogenic ability of HUVECs mediated by Rictor over-expression adenovirus.Methods The Rictor was obtained through PCR and cloned into GV314 plasmid to construct recombinant plasmid,then co-transfected 293T cells with helper plasmids to obtain Rictor overexpressing adenoviral particles(Ad-Rictor),the vector without target gene Ad-Null was set as the negative control group.Ad-Rictor and Ad-Null were infected HUVECs respectively,we also set up blank control group and resveratrol-intervention group(Ad-Rictor+Res).The expression of recombinant protein was detected by fluorescence microscopy and Western blot.CCK-8 assay,wound healing and matrigel assay were performed to assess the proliferation,migration and tube formation of HUVECs.Results We constructed Ad-Rictor and Ad-Null which may infect HUVECs and express Rictor protein efficiently.Ad-Rictor could significantly improve the proliferation,migration and lumen formation(P<0.05),resveratrol intervention may significantly inhibit these functions induced by Ad-Rictor(P<0.05).Conclusions Resveratrol inhibits the proliferation,migration and angiopoietic ability of HUVECs through targeting mTORC2/Rictor.

关 键 词：白藜芦醇 静脉移植物再狭窄 mTORC2 RICTOR 

分 类 号：R318.11[医药卫生—生物医学工程]