Tubastatin A对猪心脏停搏复苏后脑损伤时细胞焦亡的影响
Effect of tubastatin A on pyroptosis during brain injury after cardiac arrest and resuscitation in swine
陈启江 1徐杰丰 2王才木 1吴新杰 3赵雪4
作者信息
- 1. 宁海县第一医院重症医学科,宁海 315600
- 2. 浙江大学医学院附属第二医院急诊医学科,杭州 310009
- 3. 宁海县第一医院急诊医学科,宁海 315600
- 4. 浙江大学医学院附属杭州市第一人民医院急诊医学科,杭州 310006
- 折叠
摘要
目的 评价Tubastatin A(TubA)对猪心脏停搏复苏后脑损伤时细胞焦亡的影响.方法 普通级雄性白猪22头,体质量34～39 kg,4～6月龄,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)和心脏停搏复苏+TubA组(CA-CPR+TubA组,n=8).CA-CPR组和CA-CPR+TubA组采用心脏停搏9 min、心肺复苏6 min的方法制备猪心脏停搏复苏模型.CA-CPR+TubA组于复苏成功后5 min时,经1 h股静脉泵注TubA 4.5 mg/kg(用50 ml生理盐水溶解).于造模前及复苏成功后1、2、4和24 h(T0-T4)时,采集股静脉血标本,采用ELISA法检测血清NSE和S100β蛋白浓度.于T4时行神经功能缺损评分(NDS评分),随后处死猪,取大脑皮层组织,采用 Western blot 法检测组蛋白去乙酰化酶 6(HDAC6)、caspase-3、活化型 caspase-3(cleaved caspase-3)、消皮素E(GSDME)和N端GSDME(N-GSDME)的表达,ELISA法检测高迁移率族蛋白B1(HMGBl)、IL-1β和IL-18的含量.结果 与S组比较,CA-CPR组和CA-CPR+TubA组T1-T4时血清NSE和S100β蛋白浓度升高,T4时NDS评分升高,大脑皮层组织HDAC6、caspase-3、cleaved caspase-3、GSDME 和 N-GSDME 表达上调,HMGB1、IL-1β 和 IL-18 含量升高(P＜0.05);与 CA-CPR 组比较,CA-CPR+TubA组T3和T4时血清NSE和S100β蛋白浓度降低,T4时NDS评分降低,大脑皮层组织HDAC6、caspase-3、cleaved caspase-3、GSDME 和 N-GSDME 表达下调,HMGB1、IL-1β 和 IL-18 含量降低.结论 TubA减轻猪心脏停搏复苏后脑损伤的机制可能与抑制细胞焦亡有关.
Abstract
Objective To evaluate the effect of tubastatin A(TubA)on pyroptosis during brain in-jury after cardiac arrest and resuscitation in swine.Methods Twenty-two conventional male white swine,weighing 34-39 kg,aged 4-6 months,were divided into 3 groups using a random number table:sham op-eration group(group S,n=6),cardiac arrest-cardiopulmonary resuscitation(CA-CPR)group(n=8)and CA-CPR+TubA group(n=8).The swine model of CA-CPR was established by 9 min of cardiac arrest and 6 min of cardiopulmonary resuscitation in CA-CPR group and CA-CPR+TubA group.TubA 4.5 mg/kg(in 50 ml of normal saline)was infused over 1 h via the femoral vein starting from 5 min after resuscitation in CA-CPR+TubA group.Before developing the model and at 1,2,4 and 24 h after resuscitation(T0-4),blood samples were collected from the femoral vein for determination of the concentrations of neuron specific enolase(NSE)and S100β protein in serum(by enzyme-linked immunosorbent assay).Neurological deficit score(NDS)was evaluated at T4.The animals were then sacrificed,and their brain cortex tissues were har-vested to measure the expression of histone deacetylase 6(HDAC6),caspase-3,cleaved caspase-3,gas-dermin E(GSDME)and GSDME N-terminal(N-GSDME)(by Western blot)and contents of high mobility group box 1(HMGB1),interleukin-1β(IL-1β)and IL-18(by enzyme-linked immunosorbent assay).Re-sults Compared with group S,the serum concentrations of NSE and S100β were significantly increased at T1-4,NDS was increased at T4,the expression of HDAC6,caspase-3,cleaved caspase-3,GSDME and N-GSDME in brain cortex was up-regulated,and the contents of HMGB1,IL-1β and IL-18 were increased in CA-CPR and CA-CPR+TubA groups(P＜0.05).Compared with group CA-CPR,the serum concentrations of NSE and S100β were significantly decreased at T3.4,NDS was decreased at T4,the expression of HDAC6,caspase-3,cleaved caspase-3,GSDME and N-GSDME in brain cortex was down-regulated,and the contents of HMGB1,IL-1β and IL-18 were decreased in group CA-CPR+TubA(P＜0.05).Conclusions The mechanism by which TubA alleviates brain injury after cardiac arrest and resuscitation may be related to inhibition of pyroptosis in swine.
关键词
心脏停搏/心肺复苏术/再灌注损伤/脑/细胞焦亡/Tubastatin/AKey words
Heart arrest/Cardiopulmonary resuscitation/Reperfusion injury/Brain/Py-roptosis/Tubastatin A引用本文复制引用
基金项目
宁波市自然科学基金(2022J051)
浙江省医药卫生科技计划(2020KY907)
浙江省医药卫生科技计划(2023KY305)
浙江省医药卫生科技计划(2023KY924)
宁波市科技计划(2021S123)
出版年
2024
NETL
NSTL
维普
万方数据