纳布啡通过激活MAPK/ERK信号通路减轻心肌缺血再灌注大鼠心脏损伤  

Activation of MAPK/ERK pathway by nalbuphine attenuates cardiac in⁃jury in rats with myocardial ischemia-reperfusion

作　　者：李涛[1] 袁振武[1] 王义凤[1] 吴仪[1] LI Tao;YUAN Zhen-wu;WANG Yi-feng;WU Yi(Department of Anesthesiology,Jingmen First People's Hospital,Jingmen 448001,China)

机构地区：[1]荆门市第一人民医院麻醉科,湖北荆门448001

出　　处：《中国病理生理杂志》2022年第5期795-801,共7页Chinese Journal of Pathophysiology

基　　金：荆门市引导性科研计划项目(No.2018YDKY044)。

摘　　要：目的:探究纳布啡(NAL)对心肌缺血再灌注(MIR)大鼠心脏损伤的影响及其与丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号通路的关系。方法:经在体冠脉阻断法构建MIR大鼠模型,随机分为MIR组、NAL组和NAL+PD98059[MAPK激酶1/2(MEK1/2)抑制剂]组,另设假手术(sham)组,每组10只。系活结30 min后,各组大鼠给予相应的生理盐水、2.5 mg/kg NAL和10 mg/kg PD98059腹腔注射。2,3,5-三苯基氯化四氮唑(TTC)、HE及TUNEL染色分别检测心肌梗死面积、病理变化及细胞凋亡情况;试剂盒检测血清肌酸激酶(CK)、心肌钙蛋白T(cTnT)和乳酸脱氢酶(LDH)水平,以及心肌丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)和总抗氧化能力(T-AOC)水平;Western blot检测心肌凋亡相关蛋白及MAPK/ERK信号通路相关蛋白水平。结果:相比于sham组,MIR组心肌纤维断裂,心肌细胞变形,可见炎症细胞浸润,心肌梗死面积增大,细胞凋亡率,心肌cleaved caspase-3蛋白水平,血清CK、cTnT和LDH水平,以及心肌MDA和ROS水平显著升高(P<0.05),而心肌SOD和T-AOC水平及Raf、MEK1/2和ERK1/2磷酸化水平显著降低(P<0.05)。相比于MIR组,NAL干预可减轻心肌病理损伤,缩小心肌梗死面积,降低细胞凋亡率和心肌cleaved caspase-3蛋白水平,降低血清CK、cTnT和LDH水平及心肌MDA和ROS水平(P<0.05),升高SOD和T-AOC水平及Raf、MEK1/2和ERK1/2磷酸化水平(P<0.05),而PD98059能在一定程度上减弱NAL的保护作用。结论:NAL可能通过激活MAPK/ERK信号通路调节氧化应激和凋亡,从而减轻MIR导致的心脏损伤。AIM:To explore the effect of nalbuphine(NAL)on the heart injury of myocardial ischemia-reperfusion(MIR)rats and its relationship with mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK)signaling pathways.METHODS:The MIR rat model was constructed by in vivo coronary artery occlusion,and then the rats were randomly divided into MIR group,NAL group and NAL+PD98059[MAPK kinase 1/2(MEK1/2)inhibitor]group,while sham rats were used as controls,with 10 rats in each group.Thirty minutes after tying the slipknot,the rats in each group were intraperitoneally given normal saline,2.5 mg/kg NAL or 10 mg/kg PD98059.2,3,5-Triphenyltetrazolium chloride(TTC)staining,HE staining and TUNEL staining were used to detect the myocardial infarction area,pathological changes and cell apoptosis.Commercial kits were used to detect the serum levels of creatine kinase(CK),cardiac troponin T(cTnT)and lactate dehydrogenase(LDH),and myocardial malondialdehyde(MDA),superoxide dismutase(SOD),reactive oxygen species(ROS)and total antioxidant capacity(T-AOC)levels.Western blot was used to detect the levels of myocardial apoptosis-related proteins and MAPK/ERK signaling pathway-related proteins.RE⁃SULTS:Compared with sham group,the myocardial fibers in MIR group were broken,and myocardial cells deformed,with inflammatory cell infiltration.The myocardial infarction area,cell apoptosis rate,cleaved caspase-3 protein,serum CK,cTnT and LDH levels,and myocardial MDA and ROS levels were increased significantly(P<0.05),while myocardial SOD and T-AOC levels,and phosphorylation levels of Raf,MEK1/2 and ERK1/2 were reduced significantly(P<0.05).Compared with MIR group,NAL intervention attenuated myocardial damage,and reduced myocardial infarction area,cell apoptosis rate,and myocardial cleaved caspase-3,CK,cTnT,LDH,MDA and ROS levels(P<0.05).It also increased myocardial SOD and T-AOC levels,and phosphorylation levels of Raf,MEK1/2 and ERK1/2(P<0.05).Treatment with PD98059 partially blocked the protective effect of NAL.CONCLUSION

关 键 词：纳布啡 心肌缺血再灌注 MAPK/ERK通路 细胞凋亡 氧化应激 

分 类 号：R542.2[医药卫生—心血管疾病] R363.2[医药卫生—内科学]