Effects of autophagy on apoptosis in rat hippocampal neurons following cardiac arrest and cardiopulmonary resuscitation
Objective To explore the effect of autophagy on apoptosis in rat hippocampal neurons following cardiac arrest and cardiopulmonary resuscitation (CA/CPR).Methods Forty Sprague-Dawley (SD) rats were randomly divided into 4 groups:sham,CA/CPR model,rapamycin (CA/CPR + Rapa) and 3-Methyladenine (CA/CPR + 3-MA) groups.The CA/CPR model was established by the asphyxial method.The rapamycin (autophagy agonist) or 3-methyladenine (autophagy inhibitor) were used to treated the corresponding group,respectively.The neurological deficit score (NDS) was used to evaluate the neurological function of CA/CPR rats.The TUNEL staining method was used to detect the apoptosis rate of hippocampal neurons of CA/CPR rats.The reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method were used to detect the expression levels of microtubule-associated protein light chain 3 (LC3),Beclin-1,Bax,Bcl-2 and Caspase-3 mRNA and protein in hippocampus of CA/CPR rats.Results Compared with the sham group,the NDS of rats was decreased in the CA/CPR group;the apoptosis rate of the hippocampal neurons in CA/CPR group was increased;the expression levels of LC3,Beclin-1,Caspase-3,Bax were up-regulated,and Bcl-2 was down-regulated in the hippocampus of CA/CPR group (P ＜ 0.05,P ＜ 0.01).Compared with the model group,Rapa decreased NDS of CA/CPR rats,promoted the apoptosis rate of the hippocampal neurons,up-regulated the expression levels of LC3,Beclin-1,Caspase-3 and Bax in hippocampal tissue,and down-regulated the expression of Bcl-2;3-MA increased CA/CPR rats' NDS,inhibited the apoptosis rate of the hippocampal neurons,down-regulated the expression levels of LC3,Beclin-1,Caspase-3 and Bax in hippocampal tissue,and up-regulated the expression of Bcl-2 (P ＜ 0.05,P ＜0.01).Conclusion The increased level of autophagy may promote the apoptosis of CA/CPR rats' hippocampal neurons.The inhibited level of autophagy may suppress the apoptosis of CA/CPR rats' hippocampal neurons.The interaction of these two effects may be involved in the pathological process of CA/CPR.
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