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Kaczynski 2010 FENS Abstr

From Bioblast
Kaczynski L, Goralska J, Sliwa A, Zapala B, Czech U, Awsiuk M, Staszel T, Dembinska-Kiec A (2010) The protective effect of L-arginine in LN-18 glioblastoma cells. FENS Abstr vol.5,050.67.

Link: FENS Abstracts

Kaczynski L, Goralska J, Sliwa A, Zapala B, Czech U, Awsiuk M, Staszel T, Dembinska-Kiec A (2010)

Event:

Background: L-arginine (2-amino-5-guanidino-pentanoic acid) is an important amino acid for birds, carnivores and mammals. Its metabolism is complex and is only partially known. L-arginine is a substrate for nitric oxide (NO) which penetrates freely across cell membranes. The enhanced generation of NO may reduce necrosis and apoptosis in ischemia/reperfusion-induced injury in rat liver. NO donors also protect the ischemic heart from apoptosis and mitochondrial dysfunction via PKG-mediated blockage of mitochondrial permeability transition pores and subsequenced cytochrome c release. It is anticipated that NO through cGMP-dependent mechanisms can activate survival paths in hippocampal neurons and prevent apoptosis. Aims: The study of the mitochondria-related antiapoptotic influence of L-arginine against proapoptotic, proinflammatory, and metabolic stressor staurosporine in human glioblastoma LN-18 (brain cell line). Methods: The cultured glioblastoma LN-18 cells were preincubated with L-arginine (L-arg; 0.1, 0.3 or 1 mM) for 24 hours and were challenged with staurosporine (STS; 0.025 microM) for the last four hours of incubation. Measurement of the mitochondrial respiration rates was performed using Oxygraph-2k (OROBOROS®). The ATP generation was followed by using luminescence method with Luciferase/Luciferine (ATP Lite, Parkin Elmer) commercial kits. Additionally the changes in the internal mitochondrial membrane potential by the high-content cell analysis confocal fluorescent microscopy which allows prolonged imaging of live cells in controlled environment (BD Pathway 855 Bioimager) are presently performed with the usage of TMRM (for active mitochondria staining) and Hoechst (for nucleus staining). Results: L-arginine per se at 0.3 mM to 1 mM increased ATP generation, but this effect was reduced by the presence of the proapoptotic staurosporine. Conclusions: L-arginine seems to increase ATP generation in LN-18 cells, however this effect could not overcome the influence of staurosporine. The possible influence on mitochondrial membrane potential will be presented.

Keywords: Neurotoxicity, inflammation, neuroprotection, human glioblastoma LN-18 (brain cell line)

O2k-Network Lab: PL Krakow Dembinska-Kiec A


Labels: MiParea: Respiration 

Stress:Ischemia-reperfusion  Organism: Human  Tissue;cell: Neuroblastoma  Preparation: Intact cells  Enzyme: Complex V;ATP synthase  Regulation: ATP production 


HRR: Oxygraph-2k