Effects of oxygen and glucose deprivation-reperfusion on autophagy and apoptosis of PC12 cells

CHEN Ai; CHEN Zhi; SHEN Xing

doi:10.3969/j.issn.1002-266X.2016.44.006

Abstract：Objective To observe the effects of oxygen and glucose deprivation-reperfusion ( OGD/OGD-R) on auto-phagy and apoptosis of PC12 cells, and to evaluate the effective time window of OGD/OGD-activated autophagy-regulated neuron injury repair .Methods The high differentiated PC12 cells were cultured under hypoxia (1%O2 , 5%CO2 , 94%N2) and then randomly divided into 4 groups which were respectively cultured for 0 h (OGD 0 h group), 1 h(OGD 1 h group), 2 h (OGD 2 h group), 3 h (OGD 3 h group), followed by persistent culture in high glucose Dulbecco′s modified Eagle′s medium (DMEM) which supplemented with 10 %fetal bovine serum for another 0, 2, 4, 8, 12 h, respectively. The cell viability was analyzed by CCK-8.The expression of microtubule-associated protein 2-light chain 3 ( LC3Ⅱ) and Caspase-3 gene of PC12 cells in each group at different time was detected RT-PCR.Results At each oxygen-glucose reperfusion time point , the cell viability of OGD 1 h group was higher than that of OGD 2 h and OGD 3 h group, the differ-ence was statistically significant between these groups (all P<0.05).The cell viability of each OGD group gradually de-creased as the reperfusion duration till to 4, 8 and 12 h, the difference was statistically significant between these time points (all P<0.05), except the comparison between 0 h and 4 h (all P>0.05).The relative expression of LC3Ⅱgene at reperfusion of 4 h ( R4H) was lower than that of 0 h in each reperfusion groups including the control group .The relative ex-pression of Caspase-3 gene at R4H was lower than that of R0 h in OGD 1 h and OGD 2 h groups respectively .The relative expression of LC3Ⅱand Caspase-3 gene at R12H in the OGD 1 h and OGD 2 h groups was higher than that at R 4H and R8H, and the difference between groups and within groups was statistically significant (all P<0.05).Conclusions OGD-R can induce the autophagy and apoptosis of PC 12 cells.The penumbra repair time window is advised to be less than 4 hours after oxygen and glucose deprivation .