Effect of Iduna protein overexpression on PARP-1/AIF pathway during oxygen-glucose deprivation-induced damage to hippocampal neurons in neonatal rats

Abstract

Objective To evaluate the effect of Iduna protein overexpression on poly(ADP-ribose) polymerase 1 (PARP-1)/apoptosis-inducing factor (AIF) pathway during oxygen-glucose deprivation (OGD)-induced damage to hippocampal neurons in neonatal rats. Methods Primarily cultured hippocampal neurons of healthy Sprague-Dawley rats born within 24 h were isolated and cultured.Hippocampal neurons were divided into 4 groups (n=40 each) using a random number table: control group (group C), OGD group (group O), negative control lentivirus group (group NL) and Iduna protein overexpression group (group IO). OGD was induced by incubating the neurons in glucose-free medium and hypoxic environment.Negative control lentivirus and recombinant lentivirus overexpressing Iduna were added at 48 h before establishing the model in NL and IO groups, respectively.Neurons were cultured in the normal culture medium for 24 h in group C. The cell survival rate was detected using methyl thiazolyl tetrazolium assay, the lactic dehydrogenase (LDH) leakage rate was measured using colorimetric method, the comet assay was used to detect DNA content in the tail of neurons, and the expression of PARP-1, cytochrome C (Cyt c) and AIF in the nucleus was detecteded by Western blot. Results Compared with group C, the survival rate of neurons was significantly decreased, the LDH leakage rate and DNA content in the tail of neurons were increased, and the expression of PARP-1, Cyt c and AIF was up-regulated in the other three groups (P 0.05). Compared with the group NL, the survival rate of neurons was significantly increased, and the LDH leakage rate and DNA content in the tail of neurons were decreased, and the expression of PARP-1, AIF and Cyt c was down-regulated in the group IO (P<0.05). Conclusion Iduna protein overexpression reduces OGD-induced damage to hippocampal neurons through inhibiting PARP-1/AIF pathway in neonatal rats. Key words: Poly(ADP-ribose) polymerases; Apoptosis inducing factor; Ubiquitin-protein ligases; Hypoxia-ischemia, brain; Infant, newborn; Hippocampus; Neurons