Effects of autophagy on apoptosis in rat hippocampal neurons following cardiac arrest and cardiopulmonary resuscitation

doi:10.16098/j.issn.0529-1356.2018.01.006

Abstract

Abstract:

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.

Key words: Cardiac arrest, Cardiopulmonary resuscitation, Hippocampal neuron, Autophagy, Western blotting, Rat

CLC Number:

Ｒ605. 974

MA Lei WANG Rong LI Hai-long MA Xiao-long CHEN Wei YANG Li-shan. Effects of autophagy on apoptosis in rat hippocampal neurons following cardiac arrest and cardiopulmonary resuscitation[J]. Acta Anatomica Sinica, 2018, 49(1): 35-40.

References

［1］Forgacs PB, Frey HP, Velazquez A, et al. Dynamic regimes of neocortical activity linked to corticothalamic integrity correlate with outcomes in acute anoxic brain injury after cardiac arrest ［J］. Ann Clin Transl Neurol, 2017, 4(2): 119-129.

［2］Elmer J, Callaway CW. The brain after cardiac arrest ［J］. Semin Neurol, 2017, 37(1): 19-24.

［3］Talikowska M, Tohira H, Bailey P, et al. Cardiopulmonary resuscitation quality: Widespread variation in data intervals used for analysis ［J］. Resuscitation, 2016, 102(5): 25-28.

［4］Fernandez A, Solis A, Canete P, et al. Incidence and prognosis of nosocomial infection after recovering of cardiac arrest in children ［J］. Resuscitation, 2017, 113(2): 87-89.

［5］Lei LY, Sun LL, Xie L, et al. Effect of edaravone on prolonged survival and neurological function in a rat model of cardiopulmonary resuscitation after cardiac arrest ［J］. Shandong Medical Journal, 2013, 53(20): 4-6，10. (in Chinese)

雷玲艳，孙雷雷，谢露，等. 依达拉奉对心脏骤停大鼠心肺复苏后远期生存及神经功能的影响［J］. 山东医药，2013，53（20）：4-6，10.

［6］Gao L, Jauregui CE, Teng Y. Targeting autophagy as a strategy for drug discovery and therapeutic modulation ［J］. Future Med Chem, 2017, 9(3): 335-345.

［7］Wang HJ, Tan YZh. Mechanism of autophagy opens a new way for treatment of diseases ［J］. Acta Anatomica Sinica, 2017, 48(1):103-105. (in Chinese)

王海杰，谭玉珍. 细胞自噬机制开启疾病治疗新途径［J］. 解剖学报，2017，48（1）：103-105.

［8］Maino B, Paparone S, Severini C, et al. Drug target identification at the crossroad of neuronal apoptosis and survival ［J］. Expert Opin Drug Discov, 2017, 12(3): 249-259.

［9］Cui D, Shang H, Zhang X, et al. Cardiac arrest triggers hippocampal neuronal death through autophagic and apoptotic pathways ［J］. Sci Rep, 2016, 6(11): 1-14.

［10］Sun L, Guo Y, Ren PCh, et al. Neuroprotective effects of sevoflurane postconditioning on globalcerebral ischemic injury after cardiopulmonary resuscitation following asphyxia cardiac arrest in rats ［J］. Journal of Clinical Anesthesiology, 2015, 31(3): 290-294. (in Chinese)

孙丽，郭瑜，任鹏程，等. 七氟醚后处理对大鼠窒息性心脏骤停心肺复苏后全脑损伤的保护作用［J］. 临床麻醉学杂志，2015，31（3）：290-294.

［11］Xu M, Zhou GM, Wang LH, et al. Inhibiting high-mobility group box 1 (hmgb1) attenuates inflammatory cytokine expression and neurological deficit in ischemic brain injury following cardiac arrest in rats ［J］. Inflammation, 2016, 39(4): 1594-1602.

［12］Reis C, Akyol O, Araujo C, et al. Pathophysiology and the monitoring methods for cardiac arrest associated brain injury ［J］. Int J Mol Sci, 2017, 18(1): 1-18.

［13］Xiang Y, Zhao H, Wang J, et al. Inflammatory mechanisms involved in brain injury following cardiac arrest and cardiopulmonary resuscitation ［J］. Biomed Rep, 2016, 5(1): 11-17.［14］Liang P, Le W. Role of autophagy in the pathogenesis of multiple sclerosis ［J］. Neurosci Bull, 2015, 31(4): 435-444.

［15］Chen YH, Li HW, Tong XT, et al. Expression of the autophagy-related protein in hippocampus in a rat model of autism ［J］. Acta Anatomica Sinica, 2014, 45(6): 768-772. (in Chinese)

陈运华，李宏伟，童雪涛，等. 孤独症大鼠海马自噬相关蛋白的表达［J］. 解剖学报，2014，45（6）：768-772.

［16］Ding TB, Jian LG, Zhang LH, et al. Effect of rosuvastatin on expression of Beclin-1 in myocardial tissue of rats with myocardial hypertrophy ［J］. Journal of Zhengzhou University(Medical Sciences), 2016, 51(2): 251-254. (in Chinese)

丁同斌，简立国，张丽华，等. 瑞舒伐他汀对心肌肥厚大鼠心肌组织Beclin-1蛋白表达的影响［J］. 郑州大学学报(医学版)，2016，51（2）：251-254.

［17］Zhai XL, Chen MH, Xie L. Effects of tea polyphenol on neuronal injury and apoptosis in the hippocampal region of rats after cardiopulmonary resuscitation ［J］. Journal of Guangxi Medical University, 2012, 29(2): 165-168. (in Chinese)

瞿雪莉，陈蒙华，谢露. 茶多酚对心脏骤停大鼠复苏后海马神经元损伤及凋亡的影响［J］. 广西医科大学学报，2012，29（2）：165-168.

［18］Chen B, Wang G, Li W, et al. Memantine attenuates cell apoptosis by suppressing the calpain-Caspase-3 pathway in an experimental model of ischemic stroke ［J］. Exp Cell Res, 2017, 351(2): 163-172.

［19］Xi H, Fan X, Zhang Z, et al. Bax and Bcl-2 are involved in the apoptosis induced by local testicular heating in the boar testis ［J］. Reprod Domest Anim, 2017, 52(3):359-365.

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