TY - JOUR
T1 - Inhibition of prolyl hydroxylase domain-containing protein suppressed lipopolysaccharide-induced TNF-α expression
AU - Takeda, Kotaro
AU - Ichiki, Toshihiro
AU - Narabayashi, Eriko
AU - Inanaga, Keita
AU - Miyazaki, Ryohei
AU - Hashimoto, Toru
AU - Matsuura, Hirohide
AU - Ikeda, Jiro
AU - Miyata, Toshio
AU - Sunagawa, Kenji
PY - 2009/12
Y1 - 2009/12
N2 - OBJECTIVE-: Prolyl hydroxylase domain-containing proteins (PHDs) play pivotal roles in oxygen-sensing system through the regulation of α-subunit of hypoxia-inducible factor (HIF), a key transcription factor governing a large set of gene expression to adapt hypoxia. Although tissue hypoxia plays an essential role in maintaining inflammation, the role of PHDs in the inflammatory responses has not been clearly determined. Here, we investigated the role of PHDs in lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) induction in macrophages. METHODS AND RESULTS-: Northern blot analysis and ELISA revealed that LPS-induced TNF-α upregulation was strongly suppressed by PHD inhibitors, dimethyloxallyl glycine (DMOG), and TM6008 in RAW264.7 macrophages. DMOG suppressed LPS-induced TNF-α upregulation in HIF-1α-depleted cells and HIF-1α overexpression failed to suppress the induction of TNF-α. DMOG rather suppressed LPS-induced NF-κB transcriptional activity. Downregulation of Phd1 or Phd2 mRNA by RNA interference partially attenuated LPS-induced TNF-α induction. DMOG also inhibited LPS-induced TNF-α production in peritoneal macrophages as well as human macrophages. CONCLUSIONS-: PHD inhibition by DMOG or RNA interference inhibited LPS-induced TNF-α upregulation in macrophages possibly through NF-κB inhibition, which is independent of HIF-1α accumulation. This study suggests that PHDs are positive regulators of LPS-induced inflammatory process, and therefore inhibition of PHD may be a novel strategy for the treatment of inflammatory diseases.
AB - OBJECTIVE-: Prolyl hydroxylase domain-containing proteins (PHDs) play pivotal roles in oxygen-sensing system through the regulation of α-subunit of hypoxia-inducible factor (HIF), a key transcription factor governing a large set of gene expression to adapt hypoxia. Although tissue hypoxia plays an essential role in maintaining inflammation, the role of PHDs in the inflammatory responses has not been clearly determined. Here, we investigated the role of PHDs in lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) induction in macrophages. METHODS AND RESULTS-: Northern blot analysis and ELISA revealed that LPS-induced TNF-α upregulation was strongly suppressed by PHD inhibitors, dimethyloxallyl glycine (DMOG), and TM6008 in RAW264.7 macrophages. DMOG suppressed LPS-induced TNF-α upregulation in HIF-1α-depleted cells and HIF-1α overexpression failed to suppress the induction of TNF-α. DMOG rather suppressed LPS-induced NF-κB transcriptional activity. Downregulation of Phd1 or Phd2 mRNA by RNA interference partially attenuated LPS-induced TNF-α induction. DMOG also inhibited LPS-induced TNF-α production in peritoneal macrophages as well as human macrophages. CONCLUSIONS-: PHD inhibition by DMOG or RNA interference inhibited LPS-induced TNF-α upregulation in macrophages possibly through NF-κB inhibition, which is independent of HIF-1α accumulation. This study suggests that PHDs are positive regulators of LPS-induced inflammatory process, and therefore inhibition of PHD may be a novel strategy for the treatment of inflammatory diseases.
KW - Hypoxia
KW - Hypoxia-inducible factor
KW - Inflammation
KW - Prolyl hydroxylase domain-containing protein
KW - Tumor necrosis factor-alpha
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U2 - 10.1161/ATVBAHA.109.196071
DO - 10.1161/ATVBAHA.109.196071
M3 - Article
C2 - 19762779
AN - SCOPUS:73849101811
VL - 29
SP - 2132
EP - 2137
JO - Arteriosclerosis, Thrombosis, and Vascular Biology
JF - Arteriosclerosis, Thrombosis, and Vascular Biology
SN - 1079-5642
IS - 12
ER -