自噬水平变化对心脏骤停心肺复苏后大鼠海马神经元凋亡的影响

doi:10.16098/j.issn.0529-1356.2018.01.006

解剖学报 ›› 2018, Vol. 49 ›› Issue (1): 35-40.doi: 10.16098/j.issn.0529-1356.2018.01.006

自噬水平变化对心脏骤停心肺复苏后大鼠海马神经元凋亡的影响

马磊¹ 王蓉² 李海龙³ 马晓龙¹陈伟¹杨立山^1*

1.宁夏医科大学总医院急诊科，银川 750004; 2.宁夏医科大学总医院核医学科，银川 750004; 3.宁夏回族自治区第3人民医院内科，银川 750021

收稿日期:2017-02-27 修回日期:2017-03-30 出版日期:2017-02-06 发布日期:2018-02-06
通讯作者: 杨立山 E-mail:maleijzk@163.com
基金资助:
PI3K/Akt/mTOR通路调控大鼠心肺复苏后神经元自噬的机制研究

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

MA Lei¹WANG Rong² LI Hai-long³MA Xiao-long¹CHEN Wei¹ YANG Li-shan ^1*

1.Department of Emergency, General Hospital of Ningxia Medical University, Yinchuan 750004, China; 2.Department of Nuclear Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, China; 3.Department of Internal Medicine, the Third People’s Hospital of Ningxia, Yinchuan 750021, China

Received:2017-02-27 Revised:2017-03-30 Online:2017-02-06 Published:2018-02-06
Contact: YANG Li-shan E-mail:maleijzk@163.com

摘要/Abstract

摘要：

目的探讨自噬水平变化对心脏骤停心肺复苏（CA/CPR）后大鼠海马神经元凋亡的影响。方法将40只大鼠随机分为假手术组（sham）、CA/CPR模型组（model）、雷帕霉素（Rapa）组（CA/CPR +Rapa）及3-甲基腺嘌呤（3-MA）组（CA/CPR +3-MA）。采用呼气末夹闭气管窒息法复制大鼠CA/CPR动物模型，分别给予自噬激动剂Rapa 0.2 mg/kg及自噬抑制剂3-MA 10 mg/kg进行干预。采用神经功能缺陷评分（NDS）评价CA/CPR大鼠神经功能；用TUNEL染色法检测大鼠海马神经元的凋亡变化；用RT-PCR和Western blotting法检测大鼠海马内微管蛋白轻链3（LC3）、Beclin-1、Bax、Bcl-2及Caspase-3 mRNA和蛋白的表达水平。结果与假手术组比较，CA/CPR模型组大鼠NDS评分明显降低；海马神经元TUNEL染色阳性细胞数明显增多，凋亡率显著升高；海马内LC3、Beclin-1、Caspase-3、Bax表达上调，Bcl-2表达下调（P<0.05，P<0.01）。与模型组比较，CA/CPR+Rapa大鼠NDS评分明显降低，而海马神经元凋亡率明显有所增加，海马内LC3、Beclin-1、Caspase-3、Bax表达明显上调，而Bcl-2表达则明显有所下降；CA/CPR +3-MA大鼠NDS评分明显升高，而海马神经元凋亡率下降，海马内LC3、Beclin-1、Caspase-3、Bax表达明显下调，而Bcl-2表达则有所升高（P<0.05，P<0.01）。结论 CA/CPR后自噬水平升高促进海马神经元凋亡，自噬水平降低抑制海马神经凋亡，两者相互作用共同参与CA/CPR的病理过程。

关键词: 心脏骤停, 心肺复苏, 海马神经元, 自噬, 免疫印迹法, 大鼠

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

中图分类号:

Ｒ605. 974

马磊王蓉李海龙马晓龙陈伟杨立山. 自噬水平变化对心脏骤停心肺复苏后大鼠海马神经元凋亡的影响[J]. 解剖学报, 2018, 49(1): 35-40.

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.

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自噬水平变化对心脏骤停心肺复苏后大鼠海马神经元凋亡的影响

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

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