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The Pharmaceutical Society of Korea (대한약학회)
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Protection of LLC-PK1 Cells Against Hydrogen Peroxide­Induced Cell Death by Modulation of Ceramide Level

  • Yoo Jae Myung (College of Pharmacy, Chungbuk National University) ;
  • Lee Youn Sun (College of Pharmacy, Chungbuk National University) ;
  • Choi Heon Kyo (College of Pharmacy, Chungbuk National University) ;
  • Lee Yong Moon (College of Pharmacy, Chungbuk National University) ;
  • Hong Jin Tae (College of Pharmacy, Chungbuk National University) ;
  • Yun Yeo Pyo (College of Pharmacy, Chungbuk National University) ;
  • Oh Seik Wan (College of Medicine, Ewha Womans University) ;
  • Yoo Hwan Soo (College of Pharmacy, Chungbuk National University)
  • Published : 2005.03.01
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Abstract

Oxidative stress has been reported to elevate ceramide level during cell death. The purpose of the present study was to modulate cell death in relation to cellular glutathione (GSH) level and GST (glutathione S-transferase) expression by regulating the sphingolipid metabolism. LLC­PK1 cells were treated with H$_2$O$_2$ in the absence of serum to induce cell death. Subsequent to exposure to H$_2$O$_2$, LLC-PK1 cells were treated with desipramine, sphingomyelinase inhibitor, and N-acetylcysteine (NAC), GSH substrate. Based on comparative visual observation with H202-treated control cells, it was observed that 0.5 $\mu$M of desipramine and 25 $\mu$M of NAC exhibited about 90 and $95\%$ of cytoprotection, respectively, against H$_2$O$_2$-induced cell death. Desipramine and NAC lowered the release of LDH activity by 36 and $3\%$ respectively, when compared to $71\%$ in H$_2$O$_2$-exposed cells. Cellular glutathione level in 500 $\mu$M H202-treated cells was reduced to 890 pmol as compared to control level of 1198 pmol per mg protein. GST P1-1 expression was decreased in H$_2$O$_2$-treated cells compared to healthy normal cells. In conclusion, it has been inferred that H$_2$O$_2$-induced cell death is closely related to cellular GSH level and GST P1-1 expression in LLC-PK1 cells and occurs via ceramide elevation by sphingomyelinase activation.

Keywords

References

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