Protective effect of endothelin antagonist (TAK-044) on neuronal cell viability in in vitro oxygen-glucose deprivation model of stroke.

The present study was carried to investigate the effect of endothelin antagonist (TAK-044) in an in vitro model of stroke using primary neuronal culture. Hypoxia in neuronal culture was induced for 3 h using oxygen glucose deprivation (OGD) model, thereafter cells were reperfused. In separate group cultures were incubated with graded concentrations of TAK-044 (0.01, 0.1 and 1 microg/microl) for different time duration i.e. 6, 12 and 24 hours after reperfusion. Percent cell viability was assessed 24 h after reperfusion using MTT assay. It was observed that percent cell viability was reduced to 13.7 +/- 0.4% in the cells under 3 h hypoxic condition as compared to the cells under normal condition (100%). TAK-044 at the concentrations of 0.1 and 1 microg/microl, but not at 0.01 microg/microl showed significant (P<0.01) improvement in the % cell viability as compared to the cells in hypoxic condition. Percent cell viability at the concentration of 0.1 and 1 microg/microl for 24 h time duration after reperfusion were 54.8 +/- 3.2% and 75.4 +/- 1.8% respectively as compared to the cells under hypoxic condition (13.7 +/- 0.4%). The results demonstrate the neuroprotective effect of TAK-044 against neuronal damage caused by hypoxia induced in neuronal culture.