Anmeidan Regulates ROS/PARP-1/AIF Pathway to Alleviate Neuronal Damage in Sleep Deprivation Model
Objective To investigate the effects of Anmeidan on hippocampal neuron apoptosis and oxidative stress in a sleep deprivation model.Methods Totally 60 rats were randomly divided into the control group,model group,Anmeidan group,and melatonin group,with 15 rats in each group.The Anmeidan group received Anmeidan at the dosage of 18.18 g·kg-1·d-1,while the melatonin group received melatonin at the dosage of 100 mg·kg-1·d-1.The control group and model group were given an equal volume of normal saline.The behavioral changes of the rats were evaluated using the Ethovision video analysis system.HE staining was used to observe the hippocampal neurons,transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons,Tunnel staining was used to observe the neuronal apoptosis,and biochemical assays were conducted to measure the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-PX)in the serum.Immunofluorescence staining was used to detect the expression levels of hippocampal B-cell lymphoma-2(Bcl-2),Bcl-2 associated X(BAX),Caspase 3,Caspase 9,and reactive oxygen species(ROS).Western Blot was used to detect the expression of poly ADP-ribose polymerase 1(PARP-1)and apoptosis-inducing factor(AIF).Results Compared with the control group,the model group showed a decrease in time in central and number of entries into central(P＜0.001,P＜0.01).The number of hippocampal neurons decreased,with a disorganized and sparse arrangement,and an increased index of neuronal apoptosis(P＜0.001).The activities of serum SOD and GSH-PX were reduced,while the MDA content was increased(P＜0.001).The expression of BAX,Caspase 3,Caspase 9,PARP-1,and AIF,as well as the ROS content in the hippocampus,were up-regulated,while the Bcl-2 protein level was down-regulated(P＜0.001,P＜0.01).Compared with the model group,the Anmeidan group and melatonin group showed an increase in time in central and number of entries into central(P＜0.01,P＜0.05).The index of neuronal apoptosis was decreased(P＜0.01),and the levels of serum SOD and GSH-PX were increased,while the MDAcontent was decreased(P＜0.001,P＜0.01,P＜0.05).The expression of BAX,Caspase 3,Caspase 9,PARP-1,and AIF,as well as the ROS content in the hippocampus were down-regulated,while the Bcl-2 protein level was up-regulated(P＜0.001,P＜0.01).Conclusion Anmeidan can improve oxidative stress damage and apoptosis levels in the sleep deprivation model,and the mechanism may be related to regulating the ROS/PARP-1/AIF signaling pathway.
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