N-甲基-D-天冬氨酸受体-丝裂原活化蛋白激酶信号通路对心脑缺血再灌注损伤后细胞凋亡的调控作用及其机制

Regulatory effect and mechanism of N-methyl-D-aspartate receptor mitogen activated protein kinase signaling pathway on apoptosis after cardiac and cerebral ischemia-reperfusion injury

目的探究N-甲基-D-天冬氨酸(NMDA)受体-丝裂原活化蛋白激酶(MAPK)信号通路对心脑缺血再灌注损伤后细胞凋亡的调控作用及其机制。方法选择健康SD大鼠40只,采用随机数字表法将其分为对照组、心脏骤停/复苏组、地卓西平马来酸盐(MK801)干预组、MEK特异性抑制剂(U0126)干预组、联合干预组,每组各8只。除对照组外,其余组大鼠建立心脏骤停-心肺复苏模型,在复苏72 h取各组血样本进行血清白介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)、肌酸激酶(CK)活性、丙二醛(MDA)测定,并测定各组大鼠脑梗死面积占比、心肌细胞凋亡率以及脑组织和心肌组织的N-甲基-D-天门冬氨酸受体1(NMDAR-1)、p38丝裂素活化蛋白激酶(p38MAPK)、磷酸化氨基末端蛋白激酶(p-JNK)、磷酸化细胞外信号调节激酶(p-ERK)蛋白表达情况。结果心脏骤停/复苏组的脑梗死面积占比、心肌细胞凋亡率高于对照组、MK801干预组、U0126干预组、联合干预组,差异有统计学意义(P<0.05)。联合干预组的脑梗死面积占比、心肌细胞凋亡率低于MK801干预组、U0126干预组,差异有统计学意义(P<0.05)。联合干预组复苏72 h后的血清IL-1β、TNF-α、CK-MB、MDA低于MK801干预组、U0126干预组、心脏骤停/复苏组,差异有统计学意义(P<0.05)。联合干预组复苏72 h后的心肌组织、脑组织NMDAR1、p38MAPK、p-JNK、p-ERK蛋白表达低于MK801干预组、U0126干预组、心脏骤停/复苏组,差异有统计学意义(P<0.05)。结论NMDA受体-MAPK信号通路可能参与介导心脑缺血再灌注损伤后细胞凋亡的调控过程,其机制是通过调节机体炎症反应实现的。

Objective To explore the regulatory effect and mechanism of N-methyl-D-aspartic acid(NMDA)receptor Mitogen activated protein kinase(MAPK)signaling pathway on apoptosis after cardiac and cerebral ischemia-reperfusion injury.Methods A total of 40 cases of healthy SD rats were selected for the study.They were divided into control group,cardiac arrest/resuscitation group,MK801 intervention group,U0126 intervention group and combined intervention group by random number table method,with 8 cases in each group.In addition to the control group,the rats in other groups established cardiac arrest cardiopulmonary resuscitation model.After 72 h of resuscitation,blood samples from each group were taken for serum interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),creatine kinase(CK)activity,malondialdehyde(MDA)were measured.The proportion of cerebral infarction area,the apoptosis rate of myocardial cells and the expression of N-methyl-D-aspartate receptor 1(NMDAR-1),p38 mitogen activated protein kinase(p38MAPK),phosphorylated amino terminal protein kinase(p-JNK)and phosphorylated extracellular signal regulated kinase(p-ERK)proteins in brain and myocardial tissue were measured.Results The proportion of cerebral infarction area and myocardial apoptosis rate in cardiac arrest/resuscitation group were higher than those of the control group,MK801 group,U0126 group and combined intervention group,and the differences were statistically significant(P<0.05).The proportion of cerebral infarction area and myocardial apoptosis rate in combined intervention group were lower than those of the MK801 group and U0126 group,and the differences were statistically significant(P<0.05).After 72 h of resuscitation,the activities of IL-1β,TNF-α,CK and MDA in combined intervention group were lower than those of the MK801 intervention group,U0126 intervention group and cardiac arrest/resuscitation group,the differences were statistically significant(P<0.05).After 72 h of resuscitation,the protein expressions of NMDAR1,p38MAPK,P-JNK and P-ERK in myocardial tissue and brain tissue in combined intervention group were lower than those of the MK801 intervention group,U0126 intervention group and CARDIAC arrest/resuscitation group,and the differences were statistically significant(P<0.05).Conclusion NMDA receptor MAPK signaling pathway may be involved in the regulation of apoptosis after cardiac and cerebral ischemia-reperfusion injury,and its mechanism is realized by regulating the body's inflammatory response.