丁酸钠对猪心脏骤停复苏后心脑损伤的保护作用及机制研究  

Protective effect and mechanism of sodium butyrate on cardio-brain injury after cardiac arrest resuscitation in pigs

作　　者：孔莹莹 谢璐涛 卢晓驰 徐杰丰[1] 周光居[1] 张茂[1] Kong Yingying;Xie Lutao;Lu Xiaochi;Xu Jiefeng;Zhou Guangju;Zhang Mao(Department of Emergency Medicine the Second Affiliated HospitaZhejiang University School of Medicine/Zhejiang Provincial Key Laboratory for Diagnosis and Treatment of Severe Trauma and Burn/Zhejiang Provincial Acute and Critical Care Clinical Medical Research Center,Hangzhou 310009;Department of Emergency Medicine,Lishui Municipal Central Hospital,Lishui 323000,China)

机构地区：[1]浙江大学医学院附属第二医院急诊医学科/浙江省严重创伤与烧伤诊治重点实验室/浙江省急危重症临床医学研究中心,杭州310009 [2]绍兴第二医院急诊科 [3]丽水市中心医院急诊医学科,浙江丽水323000

出　　处：《中华危重症医学杂志（电子版）》2023年第5期355-362,共8页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：国家自然科学基金项目(82072126);浙江省重点研发计划项目(2021C03073、2021C03036);丽水市科技计划项目(2022SJZC014)。

摘　　要：目的:探讨丁酸钠对猪心肺复苏后心脑损伤的保护效果及作用机制。方法:采用随机数字法将24头健康国产雄性大白猪分为假手术组(n=6)、心肺复苏组(n=10)和丁酸钠组(n=8)。假手术组只行气管插管和动静脉置管,不进行诱导心室颤动及心肺复苏。心肺复苏组和丁酸钠组动物采用电刺激法诱导心室颤动9 min、心肺复苏6 min的方法制备猪心脏骤停-心肺复苏模型。于复苏成功后5 min时,丁酸钠组经股静脉泵入丁酸钠75 mg/kg维持1 h。假手术组和心肺复苏组采用同样方法给予等量等渗NaCl溶液。于心脏骤停前和复苏后1、2、4和24 h时采集静脉血样,应用酶联免疫吸附测定法检测血清心肌肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)、神经元特异性烯醇化酶(NSE)和S100钙结合蛋白B(S100B)表达水平。于复苏后24 h时,应用蛋白印迹法检测动物心肌及大脑皮层组织微管相关蛋白1轻链3(LC3)、泛素结合蛋白62(p62)蛋白表达水平,并采用TUNEL法检测细胞凋亡率。结果:心肺复苏组有9头猪复苏成功,其中7头存活至24 h;丁酸钠组有7头猪复苏成功,且均存活至24 h。3组动物24 h复苏成功情况比较,差异无统计学意义(χ^(2)=3.381,P=0.184)。复苏后1、2、4、24 h,3组动物血清cTnI(F=62.736、201.265、330.083、643.538,P均<0.001)、CK-MB(F=90.380、181.548、669.657、707.485,P均<0.001)、NSE(F=92.302、125.730、590.627、1130.372,P均<0.001)和S100B(F=42.831、152.004、295.517、1023.409,P均<0.001)水平比较,差异均有统计学意义。与假手术组相比,心肺复苏组与丁酸钠组动物复苏后出现心脑损伤,表现为复苏后2、4、24 h血清cTnI、CK-MB、NSE和S100B水平均显著升高,且心肺复苏组更高(P均<0.05)。复苏后24 h时,3组动物心肌细胞及大脑皮层细胞自噬标记物LC3(F=81.671、49.204,P均<0.001)、p62(F=127.620、65.594,P均<0.001)水平及凋亡率(F=116.750、249.105,P均<0.001)比较,差异均有统计学意义Objective:To explore the protective effect and mechanism of sodium butyrate on cardio-brain injury after cardiopulmonary resuscitation in pigs.Methods:A total of 24 large white healthy male pigs were divided into a sham operation group(n=6),a cardiopulmonary resuscitation group(n=10)and a sodium butyrate group(n=8)by a random number method.In the sham operation group,only tracheal intubation and arteriovenous catheterization were performed,and induced ventricular fibrillation and cardiopulmonary resuscitation were not performed.Cardiac arrest and cardiopulmonary resuscitation models were prepared by electrical stimulation for 9 min and cardiopulmonary resuscitation for 6 min.At 5 min after successful resuscitation,the sodium butyrate group was injected 75 mg/kg of sodium butyrate through the femoral vein for 1 h.Sham operation and cardiopulmonary resuscitation groups were given the same amount of isotonic NaCl solution by the same method.Serum levels of cardiac troponin I(cTnI),creatine kinase MB(CK-MB),neuron specific enolase(NSE)and S100 calcium-binding protein B(S100B)were measured by the enzyme-linked immunosorbent assay(ELISA)before cardiac arrest and at 1,2,4,and 24 h after resuscitation.At 24 h after resuscitation,the expression levels of microtubule-associated protein 1 light chain 3(LC3)and ubiquitin-binding protein 62(p62)in the cardiac muscle and cerebral cortex tissue were detected by western-blotting,and the apoptosis rate was detected by the TUNEL method.Results:In the cardiopulmonary resuscitation group,nine pigs were successfully resuscitated,seven of which survived to 24 h.In the sodium butyrate group,seven pigs were resuscitated successfully,and all survived to 24 h.There was no significant difference in the success of 24-h resuscitation among the three groups(χ^(2)=3.381,P=0.184).At 1,2,4 and 24 h after resuscitation,serum cTnI(F=62.736,201.265,330.083,643.538;all P<0.001),CK-MB(F=90.380,181.548,669.657,707.485;all P<0.001),NSE(F=92.302,125.730,590.627,1130.372;all P<0.001)and S100B(F=42.831,1

关 键 词：丁酸钠 心脏骤停 心肺复苏 心肌损伤 脑损伤 自噬 细胞凋亡 

分 类 号：R96[医药卫生—药理学]