title: Inhibition of the NF-κB pathway by R65 ribozyme gene via adeno-associated virus serotype 9 ameliorated oxidized LDL induced human umbilical vein endothelial cell injury.
abstract: OBJECTIVE
  NF-κB signaling plays a central role in the regulation of inflammatory responses in atherosclerosis. R65 ribozyme gene suppresses activation of NF-κB pathway, therefore we studied whether R65 gene therapy can ameliorate oxidized low-density lipoprotein (ox-LDL) induced human umbilical vein endothelial cells (HUVECs) injury.
  
  
  RESULTS
  Recombinant adeno-associated virus serotype 9 (rAVV9) vector was used to transfect the R65 ribozyme gene (rAVV9-R65) into HUVECs then following ox-LDL stimulation, expression of NF-κB p65 and p50 subunits, inflammatory mediators and cell apoptosis were examined. First, rAVV9-enhanced green fluorescent protein (eGFP)-R65 at 1×10(7) v.g./cell multiplicity of infection reached a long-lasting and significant increase in R65 gene expression. Second, ox-LDL treatment led to time- and dose-dependent activation of NF-κB pathway, and enhanced inflammatory response and cell death evidenced by increased expression of nuclear NF-κB p65 and p50 subunits, greater production of tumor necrosis factor α, interleukin-6 and von willebrand factor and 20.57% increased apoptotic HUVECs. Third, over-expression of R65 gene was 2-fold increased in HUVECs attenuated ox-LDL induced unclear accumulation and expression of p65 subunit and ameliorated inflammation and cell death (all P < 0.05).
  
  
  CONCLUSIONS
  rAAV9-mediated R65 ribozyme gene transfection in cultured HUVECs effectively inhibits ox-LDL induced activation of NF-κB and production of inflammatory cytokines and prevents cell apoptosis.
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhai
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Qing-Jie
authors.initials: QJ
authors.lastname: Chen
authors.affiliation: The Institution of Clinical Research, First Affiliated Hospital of Xinjiang Medical University Urumqi, China ; Baker IDI Heart and Diabetes Institute Melbourne, Australia.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Gao
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Ma
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Bang-Dang
authors.initials: BD
authors.lastname: Chen
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Zi-Xiang
authors.initials: ZX
authors.lastname: Yu
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: Li
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiang
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Xie
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Xinjiang Medical University Urumqi, China ; Xinjiang Key Laboratory of Cardiovascular Disease Research Urumqi, China.
authors.forename: Yi-Ning
authors.initials: YN
authors.lastname: Yang
mesh_terms: D050197:Atherosclerosis; D015153:Blotting, Western; D000229:Dependovirus; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics*
chemical_list: D008077:Lipoproteins, LDL; D016328:NF-kappa B; D016337:RNA, Catalytic; C099252:oxidized low density lipoprotein
keywords: NF-κB; Ribozyme gene; adeno-associated virus; endothelial; inflammation