Functional role of NF-κB in expression of human endothelial nitric oxide synthase.
The transcription factor NF-κB has an essential role in inflammation in endothelial cells. Endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) prevents vascular inflammation. However, the molecular mechanism underlying NF-κB-mediated regulation of eNOS expression has not been clearly elucidated. We here found that NF-κB-activating stimuli, such as lipopolysaccharide, tumor necrosis factor-α (TNF-α), and interleukin-1β, suppressed eNOS mRNA and protein levels by decreasing mRNA stability, without affecting promoter activity. TNF-α-mediated suppression of eNOS expression, mRNA stability, and 3'-untranslated region (3'UTR) activity were inhibited by NF-κB inhibitors and Dicer knockdown, but not by p38 MAPK and MEK inhibitors, suggesting the involvement of NF-κB-responsive miRNAs in eNOS expression. Moreover, TNF-α increased MIR155HG expression and promoter activity as well as miR-155 biogenesis, and these increases were blocked by NF-κB inhibitors. Transfection with antagomiR-155 blocked TNF-α-mediated suppression of eNOS 3'UTR activity, eNOS mRNA and protein levels, and NO and cGMP production. These data provide evidence that NF-κB is a negative regulator of eNOS expression via upregulation of miR-155 under inflammatory conditions. These results suggest that NF-κB is a potential therapeutic target for preventing vascular inflammation and endothelial dysfunction induced by suppression of miR-155-mediated eNOS expression.
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Kyu-Sun
KS
Lee
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Joohwan
J
Kim
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Su-Nam
SN
Kwak
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Kwang-Soon
KS
Lee
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Dong-Keon
DK
Lee
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Kwon-Soo
KS
Ha
Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Moo-Ho
MH
Won
Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Dooil
D
Jeoung
Department Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea.
Hansoo
H
Lee
Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-752, South Korea.
Young-Guen
YG
Kwon
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-702, South Korea. Electronic address: ymkim@kangwon.ac.kr.
Young-Myeong
YM
Kim
D020413:3' Untranslated Regions; D053487:DEAD-box RNA Helicases / Q000235:genetics; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D035683:MicroRNAs / Q000502:physiology; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis*; D009570:Nitriles / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics; D013450:Sulfones / Q000494:pharmacology; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D054358:Withanolides / Q000494:pharmacology
D020413:3' Untranslated Regions; C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D053583:Interleukin-1beta; C524942:MIRN155 microRNA, human; D035683:MicroRNAs; D009570:Nitriles; D012333:RNA, Messenger; D013450:Sulfones; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D054358:Withanolides; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C408087:DICER1 protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases; C009684:withaferin A
3′UTR; Endothelial nitric oxide synthase; NF-κB; TNF-α; miR-155