Alda-1对猪心脏停搏复苏后心肌组织细胞铁死亡的影响

Effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine

目的评价Alda-1对猪心脏停搏复苏后心肌组织细胞铁死亡的影响。方法普通级国产健康雄性大白猪22头,体重35~43 kg,采用随机数字表法分为3组:假手术组(S组,n=6)、心脏停搏复苏组(CA-CPR组,n=8)和Alda-1组(n=8)。S组只进行动物准备,CA-CPR组和Alda-1组通过右心室电极放电诱发心脏停搏8 min、心肺复苏8 min的方法制备猪心脏停搏复苏模型。Alda-1组于复苏后5 min静脉注射Alda-10.88 mg/kg,另2组注射等量溶媒。于造模前及复苏后1、2和4 h(T_(0-3))时应用PiCCO法测定每搏输出量(SV)和全心射血分数(GEF)。于T_(0-3)及复苏后24 h(T_(4))时,经股静脉采集静脉血标本,应用ELISA法检测血清cTnI浓度。随后处死动物,取左室心肌组织,应用Western blot法检测长链脂酰辅酶A合成酶4(ACSL4)与谷胱甘肽过氧化物酶4(GPX4)的表达,普鲁士蓝染色法检测铁沉积水平,ELISA法检测4-羟基-2-壬烯醛(4-HNE)含量,比色法检测丙二醛(MDA)和还原型谷胱甘肽(GSH)含量。结果与S组比较,CA-CPR组和Alda-1组T_(1-3)时SV和GEF降低,T_(1-4)时血清cTnI浓度升高,心肌ACSL4表达上调,GPX4表达下调,铁沉积水平、4-HNE和MDA含量升高,GSH含量降低(P<0.05);与CA-CPR组比较,Alda-1组T_(2,3)时SV和GEF升高,T_(3,4)时血清cTnI浓度降低,心肌ACSL4表达下调,GPX4表达上调,铁沉积水平、4-HNE和MDA含量降低,GSH含量升高(P<0.05)。结论Alda-1可减轻猪心脏停搏复苏后心肌损伤,进而改善心功能障碍,其机制可能与抑制细胞铁死亡有关。

Objective To evaluate the effect of Alda-1 on ferroptosis in cardiomyocytes after cardiac arrest and cardiopulmonary resuscitation in swine.Methods Twenty-two healthy male white swine,weighing 35-43 kg,were divided into 3 groups using a random number table method:sham operation group(group S,n=6),cardiac arrest-cardiopulmonary resuscitation group(group CA-CPR,n=8)and Alda-1 group(n=8).The animals only underwent the general preparation in group S,and the swine model of cardiac arrest and cardiopulmonary resuscitation was developed by 8 min of electrically induced cardiac arrest through the pacing catheter in the right ventricle followed by 8 min of cardiopulmonary resuscitation in CA-CPR and Alda-1 groups.Alda-10.88 mg/kg was intravenously injected at 5 min after resuscitation in group Alda-1,and the equal volume of vehicle was administered instead in the other two groups.Stroke volume(SV)and global ejection fraction(GEF)were measured using PiCCO before developing the model and at 1,2 and 4 h after resuscitation(T_(0-3)).Venous blood samples were collected from the femoral vein to measure the concentrations of serum cardiac troponin(cTnI)by enzyme-linked immunosorbent assay at T_(0-3),and at 24 h after resuscitation(T_(4)).The animals were then sacrificed,and myocardial tissues in the left ventricle were harvested to measure the expression of acyl-CoA synthetase long-chain family member 4(ACSL4)and glutathione peroxidase 4(GPX4)(by Western blot),iron deposition(by Prussian blue staining),4-hydroxy-2-nonenal(4-HNE)content(by enzyme-linked immunosorbent assay),and malondialdehyde(MDA)and glutathione(GSH)contents(by colorimetry).Results Compared with group S,SV and GEF were significantly decreased at T_(1-3),the serum concentrations of cTnI were increased at T_(1-4),myocardial ACSL4 expression was up-regulated,GPX4 expression was down-regulated,iron deposition and contents of 4-HNE and MDA were increased,and the content of GSH was decreased in CA-CPR and Alda-1 groups(P<0.05).Compared with group CA-CPR,SV and GEF were significantly increased at T_(2-3),the serum concentrations of cTnI were decreased at T_(3-4),myocardial ACSL4 expression was down-regulated,GPX4 expression was up-regulated,iron deposition and contents of 4-HNE and MDA were decreased,and the content of GSH was increased in group Alda-1(P<0.05).Conclusions Alda-1 can alleviate myocardial injury after cardiac arrest and cardiopulmonary resuscitation in swine and further improve cardiac dysfunction,and the mechanism may be related to inhibition of cell ferroptosis.