Effects of different interventions on HT-22 hippocampal neuron injury induced by oxygen and glucose deprivation

Yang Xiaosa; Yi Tailong; Zhang Sai; Yu Zeqi; Xu Zhongwei; Tu Yue; Cheng Shixiang

doi:10.3760/cma.j.issn.1673-4777.2017.01.015

Abstract：Objective To explore the effect of insulin,estrogen,proline hydroxylase 2 (PHD-2) inhibitor IOX2 and hypoxia-inducible factor 1 α (HIF-1 α) inhibitor 2-methoxyestradiol(2-MeOE2) on HT-22 hippocampal neuron injury induced by oxygen and glucose deprivation(OGD).Methods HT-22 hippocampal neurons of rats were set up as OGD model,OGD time and reoxygenation time were determined by MTT assay.The cells were divided into blank control group,OGD group,insulin group(0.004,0.02,0.1,0.5 μmol/L),estrogen group (0.1,1,10,100 μmol/L),IOX2 group (0.001,0.01,0.1,1 μmol/L) and 2-MeOE2 group(0.05,0.5,5,50 μmol/L).The blank control group was not treated with OGD and other interventions;the OGD group had OGD model and 0.1％ dimethyl sulfoxide;the other groups had OGD models and corresponding agents during reoxygenation.Neuron injury was determined by MTT assey.Results The optimal time of OGD was 14 h,cell survival rate was (41.3 ±0.6)％.Cell doubling time at 0-3 h,3-6 h,6-9 h during reoxygenation in OGD group was significantly longer than that in blank control group[(16.8 ± 0.7) h vs (6.0 ± 1.0) h,(14.4 ± 1.8) h vs (7.8 ±2.0)h,(14.2 ± 1.1)h vs (7.2 ±0.6)h] (P ＜0.01).Cell viabilities in 0.004,0.02,0.1,0.5 μmol/L insulin groups and 0.1,1 μmol/L estrogen groups were significantly higher than those in OGD group (P ＜0.01 or P ＜ 0.05);cell vitalities in 0.05,0.5,5,50 μmol/L 2-MeOE2 groups were significantly lower than those in OGD group(P ＜0.01);cell vitalities in IOX2 groups had no significant differences compared to those in OGD group (P ＞ 0.05).Conclusions The optimal OGD time to establish OGD model was 14 h;the optimal time for drug intervention during reoxygenation was 9 h.Insulin and low concentration of estrogen can oromote cell recovery;2-MeOE2 has an adverse effect on cell growth.