Effects of docosahexaenoic acid on hippocampal neuronal apoptosis and JNK protein in rats with subarachnoid hemorrhage
Objective To investigate the potential effect and mechanism of docosahexaenoic acid(DHA)on hippocampal neuronal apoptosis and c-Jun N-terminal kinase(JNK)protein in rats with sub-arachnoid hemorrhage(SAH).Methods Forty eight SPF-grade male SD rats were randomly divided into control group,sham group,model group and DHA intervention group according to the random number table method,with 12 rats in each group.The rats in model group and DHA group were injected with autologous blood(0.3 mL)into the optic chiasma to establish the SAH model.Rats in model group were intraperitoneal-ly injected with DHA(35 mg/kg)3 hours after SAH model establishment,rats in sham operation group were injected with 0.3 mL 0.9％sodium chloride solution into the optic chiasma,and rats in control group were fed normally.Neurobehavioral function of all rats was evaluated after 24 hours.The apoptosis of neuron was observed by TUNEL staining,and the expression of phosphorylated JNK(p-JNK)and apoptosis-related pro-teins Bax and Bcl-2 was observed by Western blot.Statistical analysis was performed using GraphPad Prism 7.0 software.One-way ANOVA was used for comparison among multiple groups,and Tukey test was used for further pairwise comparison.Results(1)The differences in neurobehavioral function scores among the 4 groups of rats were statistically significant(F=103.60,P＜0.05),the neurobehavioral function scores in mod-el group(8.67±1.37)and DHA intervention group(13.67±1.51)were lower than that in control group(18.00±0.00)and sham group(17.67±0.52)(all P＜0.05),while the neurobehavioral function score in DHA intervention group was higher than that in the model group(P＜0.05).(2)The results of TUNEL stai-ning showed that there were statistical differences in the number of hippocampal neuron apoptosis among the 4 groups(F=30.76,P＜0.05),the number of hippocampal neuron apoptosis in model group(55.67±5.28)was higher than those in control group(25.83±7.06)and sham group(25.50±6.72)(both P＜0.05),the number of hippocampal neuron apoptosis in DHA intervention group(35.17±5.78)was lower than that in model group.(3)The results of Western blot showed that there were no statistical differences in the Bax pro-tein levels among the four groups(F=2.00,P＞0.05).There were statistical differences in the expression lev-els of Bcl-2,p-JNK and Bax/Bcl-2 ratio among the 4 groups(F=8.48,5.69,5.39,all P＜0.05).There was no statistical difference in Bcl-2,p-JNK protein levels and Bax/Bcl-2 ratio between the control group and sham group(all P＞0.05).The p-JNK protein levels and Bax/Bcl-2 ratio in model group((1.93±0.25),(2.05±0.86))were higher than those in sham group((1.42±0.33),(1.05±0.26))(both P＜0.05),the Bcl-2 protein level in model group(1.04±0.23)was lower than that in sham group(1.61±0.16)(P＜0.05).The p-JNK protein level and Bax/Bcl-2 ratio in DHA intervention group((1.43±0.33),(1.19± 0.30))were lower than those in model group(both P＜0.05),the Bcl-2 protein level in DHA intervention group(1.42±0.28)was higher than that in model group(P＜0.05).Conclusion DHA can reduce neuro-nal apoptosis,inhibit the activation of p-JNK and improve neurological function of SAH model rats.
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