CAJ | 학술논문

目的探讨缓激肽延迟后处理对心肺复苏后大鼠海马CA1区神经元的保护效应.方法实验在福建医科大学神经生物学中心进行.24只SD大鼠随机(随机数字法)分为假手术组(Sham组)、心搏骤停后自主循环恢复(restoration of spontaneously circulation,ROSC)组及缓激肽后处理(bradykinin postconditioning,BR-P)组.以窒息法诱导建立心搏骤停模型,在心搏停止6min后开始心肺复苏.于成功复苏后2d,对BR-P组和ROSC组大鼠分别经腹腔注射缓激肽(150 μg/kg)和等量生理盐水.心肺复苏后3d,对各组大鼠进行神经功能缺损评分(neurological deficit scores, NDS)后取材,取鼠脑进行DAPI/Bax、Bcl-2、Caspase-3免疫荧光双标染色,在共聚焦显微镜下观察各组海马CA1区神经元凋亡及Bax、Bcl-2、Caspase-3蛋白表达情况.计量资料用均数±标准差((x)±s)表示,组间比较采用单因素方差分析.结果与Sham组比较,ROSC组NDS评分降低(60.13vs.80.00,P=0.000)、CA1区凋亡神经元增多、Bax、Caspase-3蛋白表达显著增强(56.70vs.37.39,P =0.000;43.46vs.37.29,P=0.004)、而Bcl-2蛋白表达减弱(41.90vs.47.52,P=0.008).BR-P组NDS评分高于ROSC组(69.75vs.60.13,P=0.000)、凋亡神经元减少、Bax、Caspase-3蛋白表达减弱(51.34vs.56.70,P=0.046;38.90vs.43.46,P=0.028)、而Bcl-2蛋白表达增强(46.04vs.41.90,P=0.048).结论缓激肽后处理可诱导Bax、Caspase-3蛋白表达减弱,Bcl-2蛋白表达增强,减轻神经元凋亡,从而对心肺复苏后脑缺血-再灌注损伤起保护效应.
목적 탐토완격태연지후처리대심폐복소후대서해마CA1구신경원적보호효응.방법 실험재복건의과대학신경생물학중심진행.24지SD대서수궤(수궤수자법)분위가수술조(Sham조)、심박취정후자주순배회복(restoration of spontaneously circulation,ROSC)조급완격태후처리(bradykinin postconditioning,BR-P)조.이질식법유도건립심박취정모형,재심박정지6min후개시심폐복소.우성공복소후2d,대BR-P조화ROSC조대서분별경복강주사완격태(150 μg/kg)화등량생리염수.심폐복소후3d,대각조대서진행신경공능결손평분(neurological deficit scores, NDS)후취재,취서뇌진행DAPI/Bax、Bcl-2、Caspase-3면역형광쌍표염색,재공취초현미경하관찰각조해마CA1구신경원조망급Bax、Bcl-2、Caspase-3단백표체정황.계량자료용균수±표준차((x)±s)표시,조간비교채용단인소방차분석.결과 여Sham조비교,ROSC조NDS평분강저(60.13vs.80.00,P=0.000)、CA1구조망신경원증다、Bax、Caspase-3단백표체현저증강(56.70vs.37.39,P =0.000;43.46vs.37.29,P=0.004)、이Bcl-2단백표체감약(41.90vs.47.52,P=0.008).BR-P조NDS평분고우ROSC조(69.75vs.60.13,P=0.000)、조망신경원감소、Bax、Caspase-3단백표체감약(51.34vs.56.70,P=0.046;38.90vs.43.46,P=0.028)、이Bcl-2단백표체증강(46.04vs.41.90,P=0.048).결론 완격태후처리가유도Bax、Caspase-3단백표체감약,Bcl-2단백표체증강,감경신경원조망,종이대심폐복소후뇌결혈-재관주손상기보호효응.
Objective To evaluate the protective effect of delayed bradykinin post-conditioning on hippocampus CA1 neurons of rat following cardiopulmonary resuscitation.Methods A total of 24 male SD rats were randomly (random number) divided into sham group,restoration of spontaneous circulation group (ROSC) and bradykinin post-conditioning group (BR-P),using random number method.Cardiac arrest was induced by suffocation method with vecuronium bromide injection to paralyze the respiratory movement.Cardiopulmonary resuscitation was performed 6 min after cardiac arrest.Two days after ROSC,bradykinin (150 μg/kg) was administrated intraperitoneally in BR-P group and the equivalent volume of saline was injected instead in ROSC group.Three days after ROSC,neurological function of rats was evaluated with neurological deficit score (NDS).And then brain tissue was taken for DAPI/ Bax,Bcl-2 and Caspase-3 with double immunofluorescence staining and neuron apoptosis and Bax,Bcl-2 and Caspase-3 expression in CA1 region of hippocampus were assessed.The results were expressed as ((x) ± s) and one way analysis of variance was employed for comparison among groups.Results Compared with sham group,ROSC group exhibited a lower NDS score (60.13 vs.80.00,P ＜0.01),higher Bax and Caspase-3 expression in CA1 region (56.70 vs.37.39,P ＜ 0.01;43.46 vs.37.29,P ＜ 0.01) and lower expression of Bel-2 (41.90 vs.47.52,P＜0.01).Compared to ROSC group,there were higher NDS score (69.75 vs.60.13,P ＜ 0.01),lower Bax and Caspase-3 expression (51.34 vs.56.70,P ＜ 0.05;38.90 vs.43.46,P ＜ 0.05) and enhanced Bcl-2 expression (41.90 vs.47.52,P ＜ 0.05) in BR-P group.Conclusions Bradykinin postconditioning can exert neuroprotective effects on rats after CPR and the neuroprotection is likely associated with down-regulation of Bax and Caspase-3,and activation of Bacl-2,inhibiting neuron apoptosis in CA1 region.