Effects Of Low Temperature On Apoptosis And Expression Of FoxO3a In PC12 Cells Induced By Oxygen Glucose Deprivation

ObjectiveBy training oxygen glucose deprivation PC 12 cells Under the conditions of temperature reduction and testing the FoxO3a expression of oxygen glucose deprivation PC 12, elaborating low temperature may protect the cells from the PI3K/Akt/FoxO3a pathway.MethodsCulturing PC 12 cells and preparing OGD (oxygen glucose deprivation) model by the method of chemical oxygen glucose deprivation. Then the PC 12 cells were divided into 6 groups after the oxygen glucose deprivation, which that 3 experimental groups and 3 control groups.3 experimental groups were added with LY294002 reagent, and the control group was not added. The experiment group and the control group were put into 3 temperature sections for culture:①Room temperature group (37 ℃ incubator culture 24h); ②Mild hypothermia group (32 ℃ incubator culture 24h);③Deep low temperature group (27 ℃ incubator culture 24h). MTT method was used to measure the cell survival rate and apoptosis index of Hoechst 33258 staining method..Protein imprinting method was used to detect the expression of FoxO3a protein in each experimental group and control group, and the results were analyzed by SPSS17.0.Results1. MTT was used to measure cell viability:the temperature in the control group cells survival rate were higher than experimental group and had statistical significance (P< 0.05); with the decrease of temperature. In the control group cells increased the survival rate of, the difference is statistically significant (P< 0.05); while the cell survival rate was not statistically significant (P> 0.05).2. Apoptosis rate of cells was measured by Hoechst staining method. Normal temperature section:There was no significant difference between the experimental group and the control group (P>0.05). Mild hypothermia and deep low temperature section:The apoptosis rate of the experimental group was higher than that of the control group and the difference was statistically significant (P< 0.05); Comparison within the experimental group:The apoptosis rate of cells decreased with the decrease of temperature but the difference was not statistically significant (P> 0.05); Compared with the control group:The apoptosis rate of cells decreased with the decrease of temperature and the difference was statistically significant (P< 0.05).3. Expression of FoxO3a protein detected by protein imprinting method. The expression of FoxO3a protein in the experimental group was higher than that in the control group, the difference was statistically significant (P< 0.05). There was no significant difference in the expression of FoxO3a protein in the experimental group (P> 0.05), but the expression of FoxO3a decreased with the decrease of the temperature in the control group, and the expression of P was statistically significant (P< 0.05).Conclusion1. The OGD model of PC 12 cell is good for vitro ischemia and hypoxia with easy to operate, simulating the process of ischemia and hypoxia.2. Decreased temperature can affect the expression of FoxO3a in oxygen glucose deprived PC 12 cells.3. low temperature can protect cells through the PI3K/Akt/FoxO3a pathway...