Effects Of Hypoxia On Apoptosis And ROS, NO Level Of Cerebral Cortex Cells In SD Rats

ObjectiveTo observe the effects of hypoxia on apoptosis and test the levels of Nitric oxide, ROS of cerebral cortex cells in vitro which were treated with different hypoxic; further explore the connection between ROS, NO and hypoxia.MethodsSeparate cerebral cortex of the newborn Sprague-Dawley (SD) rats in 24h, and culture cerebral cortex cells in vitro plated into Poly-L-Lysine-coated 4 24-pores plates at 7×10⁵ cells/ml cell density(4 duplicate pores per group). The author design two oxygen concentration (10%, 0.5%) as hypoxia groups, and each concentration set corresponding normoxia group/control group(20% oxygen concentration). After cultured 24 hours, we put 24-pores plates into hypoxystation for 2 hours, and then detect the the cell apoptotic index and ROS levels by flow cytometer(FCM), NO levels measured by enzyme linked immunosorbent assay (ELISA ). We do the experiments three times.Results1. Effects of hypoxia on cerebral cortex apoptosis: Compared the apoptosis rate of hypoxia groups and that of normoxia groups, the result is that 10H group has a higher apoptosis rate than group 10C(p<0.05); There was no significant difference between two hypoxia groups, but there is a trend that the apoptosis rate induced by 0.5%O₂ is lower than that induced by 10%O₂.2. Effects of hypoxia on level of ROS: The levels of ROS reduced significantly in hypoxia groups compared with normoxia groups (p<0.05); There was no significant difference between two hypoxia groups.3. Effects of hypoxia on level of NO: The NO value were increased after hypoxia, but there was no significant difference; Compared with 0.5% O₂ group(p< 0.05).Conclusion1. 10%O₂ increase the apoptosis rate of Cerebral Cortex Cells in SD Rats.2. Both 10% O₂ and 0.5% O₂ reduce the production of reactive oxygen species of Cerebral Cortex Cells in SD Rats.3. 10%O₂ tend to induce Cerebral Cortex Cells to produce NO.