Objectives : Cerebrovascular diseases seriously effect patient´s quality of life, and the treatment has, its limitation to bring functional recovery unless a patient is brought to a hospital within several hours from the onset. In this study, we tried to find a way to prolong the survival cells under hypoxic conditions using cell culture.
Methods : Cultured rat glioma cells grew in normal and high glucose medium under hypoxic conditions, with or without doxycycline. Cell viability, glucose and lactic acid concentration were measured.
Results : The cells died after 1 day of culture under hypoxic conditions in normal glucose medium without doxycycline due to the total consumption of glucose. On the contrary, the cells still survived even after 3 days of culture under the same hypoxic conditions in normal glucose medium with doxycycline despite the total consumption of all glucose. For both cases, the cause of death was not, at least, due to the decrease of pH as the pH was kept neutral during the whole period of culture. In this case, doxycycline was the cell survival factor by suppressing apoptosis, which was proved by DNA fragmentation assay. On the other hand, the cells died after 60 hours of culture under hypoxic conditions in high glucose medium without doxycycline due to the decrease of pH. The cells still survived even after 70 hours of culture under the same hypoxic conditions in high glucose medium with doxycycline due to the inhibition of decreased pH. For both cases, the cause of death was not, at least, due to the consumption of total glucose because some glucose remained in the medium at the end culture.
Conclusion : Doxycycline increased cell viability both in normal glucose and high glucose medium under hypoxic conditions, which might have an application for the treatment of patients with ischemic stroke.
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