Metabolic influence on oxidative stress in astrocytes and neuronal SHSY5Y cells

Introduction:

Glucose is proposed to be the main metabolite in astrocytes neuronale cells SH-SY5Y metabolizing lactate. We measured cellular ATP, glutathione (GSH), lactate, reactive oxygen species (ROS), NAD(P)H oxidase activity and lactate dehydrogenase (LDH) activity in both cell type.

Methods:

Neuroblastoma SH-SY5Y cells were cultured with 15% fetal calf serum, 1% penicillin-streptomycin and 1% glutamine in a 5% CO2 atmosphere. Dishes were incubated with 3 ml glucose (5 mM) or lactate (5 mM) krebs ringer containing medium for 24 hours. One ml of cells was used for ROS measurement. Resting cells were ultrasonic treated to analyze ATP, GSH, and NAD(P)Hoxidases.

Results:

ATP concentration is significant diminished in Astrocytes. The ATP concentration in neuronal SH-SY5Y cells do not differ between glucose or lactate metabolism. Lactate as substrate enhanced reduced glutathione content. ROS formation is significant lower in SH-SY5Y cells comparing to astrocytes. Cytosolic NAD(P)H oxidases activity in astrocytes were 3 to 4 fold higher in astrocytes. Neuronal cells showed a lower NAD(P)H oxidase activity, a 2 to 3 fold higher breakdown of lactate in neuronale cells was observed.

Conclusions:

Glucose is substrate of choise in astrocytes, Lactate diminish ATP concentration in astrocytes, the antoxidative capacity of GSH and enhance reactive oxygen species. Lactate metabolism improves ATP concentration and the oxdative capacity in SH-SY5Y cells by more efficacy of LDH.