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

作　　者：马磊 王蓉[2] 李海龙 马晓龙 陈伟[1] 杨立山[1] 

机构地区：[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

出　　处：《解剖学报》2018年第1期35-40,共6页Acta Anatomica Sinica

基　　金：宁夏自然科学基金(NZ16269)

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。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） or3-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（ RTPCR） 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,Rapadecreased 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 ’ hip

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 

分 类 号：R605.974[医药卫生—急诊医学]