title: Delphinidin prevents hypoxia-induced mouse embryonic stem cell apoptosis through reduction of intracellular reactive oxygen species-mediated activation of JNK and NF-κB, and Akt inhibition.
abstract: Delphinidin, gallic acid, betulinic acid, and ursolic acid, which are bio-active ingredients in a variety of fruits, vegetables, and herbs, have potent antioxidant activity and various biological activities. However, it is not clear whether these bio-active ingredients can significantly contribute to the protection of embryonic stem (ES) cells from hypoxia-induced apoptosis. In the present study, hypoxia-induced ES cells apoptosis with time, which were abrogated by pretreatment with all ingredients. Hypoxia-induced ROS generation was blocked by pretreatment with all ingredients in a dose-dependent manner, with the maximum ROS scavenging effect observed for delphinidin. Hypoxia increased phosphorylation of JNK and NF-κB were blocked by pretreatment of delphinidin as well as NAC. Hypoxia decreased phosphorylation of Akt(thr308) and (ser473); these decreases were reversed by pretreatment with delphinidin or NAC. However, Akt inhibition did not affect NF-κB phosphorylation. Delphinidin attenuated the hypoxia-induced increase in Bax, cleaved caspase-9, cleaved caspase-3, and decrease in Bcl-2, which were diminished by pretreatment of Akt inhibitor. Hypoxia induced Bax translocation from the cytosol to mitochondria. Furthermore, hypoxia induced mitochondria membrane potential loss and cytochrome c release in cytosol, which were blocked by delphinidin pretreatment. Hypoxia induced cleavage of procaspase-9 and procaspase-3 which were blocked by delphinidin or SP600125, but Akt inhibitor abolished the protection effect of delphinidin. Moreover, inhibition of JNK and NF-κB abolished hypoxia-induced ES cell apoptosis and inhibition of Akt attenuated delphinidin-induced blockage of apoptosis. The results indicate that delphinidin can prevent hypoxia-induced apoptosis of ES cells through the inhibition of JNK and NF-κB phosphorylation, and restoration of Akt phosphorylation.
authors.affiliation: Department of Veterinary Physiology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-741, South Korea.
authors.forename: Bit Na
authors.initials: BN
authors.lastname: Seo
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Ryu
authors.forename: Seung Pil
authors.initials: SP
authors.lastname: Yun
authors.forename: Ji Hoon
authors.initials: JH
authors.lastname: Jeon
authors.forename: Su Shin
authors.initials: SS
authors.lastname: Park
authors.forename: Keon Bong
authors.initials: KB
authors.lastname: Oh
authors.forename: Jin-Ki
authors.initials: JK
authors.lastname: Park
authors.forename: Ho Jae
authors.initials: HJ
authors.lastname: Han
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000872:Anthocyanins / Q000494:pharmacology*; D000873:Anthracenes / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D015687:Cell Hypoxia; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D048670:MAP Kinase Kinase 4 / Q000235:genetics* / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction; D051028:bcl-2-Associated X Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D000872:Anthocyanins; D000873:Anthracenes; D000975:Antioxidants; C494827:Bax protein, mouse; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; C432165:pyrazolanthrone; D051057:Proto-Oncogene Proteins c-akt; D048670:MAP Kinase Kinase 4; D020169:Caspases; C017185:delphinidin; D010100:Oxygen; D000111:Acetylcysteine