title: Ablation of NF-kappaB expression by small interference RNA prevents the dysfunction of human umbilical vein endothelial cells induced by high glucose.
abstract: Diabetes is a major independent risk factor for cardiovascular disease and stroke. High glucose (HG) reduces endothelial cell (EC) proliferation with a concomitant increase in apoptosis. HG also induces the translocation of nuclear factor (NF)-kappaB in human umbilical vein endothelial cells (HUVECs). However, data regarding the relationship between NF-kappaB signaling and HG-induced endothelial dysfunction are limited. In the present study, we constructed an NF-kappaB-targeting RNA interference (RNAi) adenovirus vector and cultured HUVECs in 5.5, 20.5, or 30.5 mM D: -glucose or in daily alternating 5.5 or 30.5 mM D: -glucose. We assessed the effects of the NF-kappaB pathway on proliferation under HG conditions by measuring bromodeoxyuridine incorporation and conducting methyl thiazolyltetrazolium assays. We also tested apoptosis by performing flow cytometry and terminal deoxynucleotidyl transferase nick-end labeling assay. The RNAi adenovirus effectively downregulated expression of the p65 protein in HUVECs for more than 6 days. Blockage of the NF-kappaB pathway with the RNAi adenovirus substantially protected HUVECs from decreased proliferation and reduced cellular apoptosis in HG conditions. These findings may explain how hyperglycemia promotes dysfunction of ECs and could elucidate a potential new target for therapeutic interventions.
authors.affiliation: Department of Endocrinology, Fujian Medical University, Fujian, China. chengang18@yahoo.com
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Shen
authors.forename: Jin
authors.initials: J
authors.lastname: Yao
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.forename: Xu
authors.initials: X
authors.lastname: Lin
authors.forename: Yufang
authors.initials: Y
authors.lastname: Qiao
authors.forename: Tingting
authors.initials: T
authors.lastname: You
authors.forename: Fenghui
authors.initials: F
authors.lastname: Lin
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Fang
authors.forename: Xin
authors.initials: X
authors.lastname: Zou
authors.forename: Lixiang
authors.initials: L
authors.lastname: Lin
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003001:Cloning, Molecular; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D055785:Gene Knockdown Techniques; D005947:Glucose / Q000009:adverse effects / Q000494:pharmacology*; D006801:Humans; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D014471:Umbilical Veins / Q000187:drug effects / Q000378:metabolism
chemical_list: D016328:NF-kappa B; D034741:RNA, Small Interfering; D005947:Glucose