Adrenomedullin regulates cellular glutathione content via modulation of gamma-glutamate-cysteine ligase catalytic subunit expression.
Adrenomedullin (AM) participates in a wide range of physiological and pathological processes including vasorelaxation, angiogenesis, cancer promotion, and apoptosis. Recently, it has been reported that AM protects a variety of cells against oxidative stress induced by stressors such as hypoxia, ischemia/reperfusion, and hydrogen peroxide through the phosphatidylinositol 3-kinase (PI3K)-dependent pathway. However, the molecular mechanisms underlying the pathway of cell survival against hypoxic injury are largely unknown. In an effort to investigate the survival mechanism against hypoxic injury, we studied the effects of AM on cellular levels of reactive oxygen species, well-known mediators of cell death after oxidative stress, and the mechanism involved in the regulation of reactive oxygen species levels. Here, we show that AM increases gamma-glutamate-cysteine ligase (gamma-GCL) activity under both hypoxic and normoxic conditions, resulting in an up-regulation of cellular glutathione levels to more than 2-fold higher than basal expression. In addition, we demonstrate that AM induces concentration-dependent expression of the catalytic subunit of gamma-GCL (gamma-GCLC) at the mRNA and protein levels through the activation of the gamma-GCLC promoter fragment sequence from -597 to -320. However, when treated with the PI3K inhibitors, the effects of AM on gamma-GCLC expression were completely abrogated, suggesting that a PI3K pathway linked AM with the transcriptional activation of the gamma-GCLC promoter. Taken together, our data suggests that AM participates in the regulation of cellular redox status via glutathione synthesis. These results may explain, in part, the mechanism by which AM protects cells against oxidative stress.
Kim
Jee-Youn
JY
Department of Pathology, College of Medicine, Kyung Hee University, Seoul 130-701, Korea.
Yim
Ji-Hye
JH
Cho
Jin-Ho
JH
Kim
Jin-Hwan
JH
Ko
Jeong-Hun
JH
Kim
Su-Mi
SM
Park
Seungjoon
S
Park
Jae-Hoon
JH
D053607:Adrenomedullin; D017209:Apoptosis; D015153:Blotting, Western; D016923:Cell Death; D045744:Cell Line, Tumor; D002470:Cell Survival; D017353:Gene Deletion; D005721:Glutamate-Cysteine Ligase / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D000860:Hypoxia; D008156:Luciferases / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D010455:Peptides / Q000378:metabolism / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D010455:Peptides; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D053607:Adrenomedullin; D006861:Hydrogen Peroxide; D008156:Luciferases; D019869:Phosphatidylinositol 3-Kinases; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione; D010100:Oxygen