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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

Apoptosis volume 18, pages 811–824 (2013)Cite this article

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 Aktthr308 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.

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Acknowledgments

This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ009090), Rural Development Administration, the Health Technology R&D project (A120216), Ministry of Health and Welfare, and Research Settlement Fund (550-20110054) for the new faculty of SNU, Republic of Korea.

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  1. Department of Veterinary Physiology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, 151-741, South Korea

    Bit Na Seo, Jung Min Ryu, Seung Pil Yun, Ji Hoon Jeon, Su Shin Park & Ho Jae Han

  2. Animal Biotechnology Division, National Institute of Animal Science, RDA, 143-13 Seosuwon-ro, Gwonsun-gu, Suwon, 441-706, South Korea

    Keon Bong Oh & Jin-Ki Park

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  1. Bit Na Seo
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  2. Jung Min Ryu
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Correspondence to Ho Jae Han.

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B. N. Seo and J. M. Ryu contributed equally to this study.

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Seo, B.N., Ryu, J.M., Yun, S.P. et al. 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. Apoptosis 18, 811–824 (2013). https://doi.org/10.1007/s10495-013-0838-2

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Keywords

  • Delphinidin
  • Apoptosis
  • Hypoxia
  • Embryonic stem cells