Swelling and death of neuronal cells by lactic acid

Lactacidosis occurring in cerebral ischemia or trauma is a major mechanism of cytotoxic brain edema and brain damage. Respective effects of lactacidosis were currently analyzed in vitro by employment of the murine neuronal cell line, Neuro-2A, in order to obtain a better understanding of specific mechanisms underlying cell swelling and cell death in comparison with glial cells. The cells were suspended in a physiological medium in the presence of lactic acid at increasing concentrations. Levels of acidosis reaching from pH 6.8-5.6 were obtained while other parameters, such as osmolarity and electrolyte concentrations, were maintained in the physiological range. Assessment of cell swelling and cell viability using exclusion of propidium iodide was made by flow cytometry with employment of an advanced Coulter system. Swelling of Neuro-2A cells commenced once the pH in the medium was lowered to 6.8 or below. From this level downward, cell swelling was a function of the severity of acidosis and duration of exposure. For example, lactacidosis of pH 6.8 or 5.6 lasting 90 min led to an increase in cell volume to 109.5% or 159.6% of normal, respectively. Viability of the neuronal cells was 85% under control conditions. It remained in this range down to pH 6.2. At pH 5.6, however, cell viability decreased in a time-dependent fashion. At 90 min, only 48.9% of the neuronal cells were viable at pH 5.6. The swelling response and impairment of viability of the neuronal cells was compared with that of C6 glioma cells.(ABSTRACT TRUNCATED AT 250 WORDS)