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24h;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(x2=3.381,P=0.184).复苏后1、2、4、24h,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、1 130.372,P 均＜0.001)和 S100B(F=42.831、152.004、295.517、1 023.409,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.与假手术组相比,心肺复苏组与丁酸钠组动物复苏后24 h心肌组织及大脑皮层组织LC3表达均增加、p62表达均减少、细胞凋亡指数均显著升高(P均＜0.05).但与心肺复苏组相比,丁酸钠组复苏后24 h心肌组织及大脑皮层组织LC3表达均减少、p62表达均增加、细胞凋亡指数均显著降低(P均＜0.05).结论 丁酸钠对心脏骤停复苏后心脑损伤具有保护作用,机制可能与抑制细胞自噬及凋亡有关.
Abstract
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(x2=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=92.302,125.730,590.627,1 130.372;all P＜0.001)and S100B(F=42.831,152.004,295.517,1 023.409;all P＜0.001)were observed and compared in the three groups,and the differences were statistically significant.Compared with the sham operation group,the cTnⅠ,CK-MB,NSE and S100B levels in serum were significantly increased at 2,4 and 24 h after resuscitation in the cardiopulmonary resuscitation group and sodium butyrate group,which were highest in the cardiopulmonary resuscitation group(all P＜0.05).At 24 h after resuscitation,the levels of autophagy markers LC3(F=81.671,49.204;both P＜0.001)and p62(F=127.620,65.594;both P＜0.001)and the apoptosis rate(F=116.750,249.105;both P＜0.001)in cardiomyocytes and cerebral cortex cells were observed and compared in the three groups,and the differences were statistically significant.Compared with the sham operation group,the LC3 expression and apoptosis index were increased,and the p62 expression was decreased in myocardium and cerebral cortex of pigs in the cardiopulmonary resuscitation group and sodium butyrate group at 24 h after resuscitation(all P＜0.05).However,compared with the cardiopulmonary resuscitation group,the LC3 expression and apoptosis index were decreased,and the p62 expression was increased in the myocardium and cerebral cortex in the sodium butyrate group at 24 h after resuscitation(all P＜0.05).Conclusion Sodium butyrate has a protective effect on cardio-brain injury after resuscitation of cardiac arrest,and the mechanism may be related to inhibition of autophagy and apoptosis.
NETL
NSTL
维普
万方数据