Ablation of NF-kappaB expression by small interference RNA prevents the dysfunction of human umbilical vein endothelial cells induced by high glucose.
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.
Chen
Gang
G
Department of Endocrinology, Fujian Medical University, Fujian, China. chengang18@yahoo.com
Shen
Xiaoyan
X
Yao
Jin
J
Chen
Feng
F
Lin
Xu
X
Qiao
Yufang
Y
You
Tingting
T
Lin
Fenghui
F
Fang
Xiaowen
X
Zou
Xin
X
Lin
Lixiang
L
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
D016328:NF-kappa B; D034741:RNA, Small Interfering; D005947:Glucose