Role of autophagy in apoptosis in cortical neurons of rats in early phase of oxygen-glucose deprivation / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the role of autophagy in apoptosis in cortical neurons of rats in the early phase of oxygen-glucose deprivation (OGD).Methods Female Sprague-Dawley rats at 16-18 days of gestation,weighing 350-400 g,were sacrificed after being anesthetized with pentobarbital sodium.The fetal rats were obtained and the primary cortical neurons were isolated and seeded in culture plates.The neurons were then divided into 4 groups (n =7 each) using a random number table:control group (group C),OGD 30 min group,OGD 1 h group,OGD 2 h group,and OGD 3 h group,OGD + autophagy inducer rapamycin group (OGD + R group),and OGD + autophagy inhibitor 3-methyladenine (3-MA) group (OGD + 3-MA group).The neurons were incubated in sugar-free DMEM medium containing 5％ CO2 and 95％ N2 for 30 min,1,2 and 3 h in OGD 30 min,1 h,2 h,and 3 h groups,respectively.Rapamycin (final concentration of 100 nmol/L) and 3-MA (final concentration of 10 mmol/L) were added to sugar-free DMEM medium in OGD + R and OGD + 3-MA groups,respectively,and the neurons were then incubated for 1 h.The CCK-8 assay was used to detect the neuronal viability after the end of OGD.The expression of caspase-3 and LC3 Ⅱ was measured by Western blot.Results Compared with C group,the survival rates of cortical neurons were significantly decreased in the other groups,the expression of LC3 Ⅱ was up-regulated in OGD 30 min,1 h,2 h and 3 h groups,no significant change was found in caspase-3 expression in OGD + R group,and the expression of caspase-3 was up-regulated in the other groups.Compared with OGD 1 h group,the survival rates of cortical neurons were significantly decreased in OGD2 h,OGD3 h and OGD + 3-MA groups,the survival rates of cortical neurons were increased,and the expression of caspase-3 was down-regulated in OGD + R group,the expression of LC3 Ⅱ was up-regulated in OGD + 3-MA group,and the expression of LC3 Ⅱ was down-regulated in OGD2 h and 3 h groups.Conclusion Autophagy initiated in the early phase of OGD can inhibit apoptosis in cortical neurons of fetal rats.