CAJ | 학술논문

目的评价七氟醚预处理对大鼠心肌缺血再灌注时wnt/糖原合成酶激酶-3β(GSK-3β)/β-连环蛋白(β-catenin)信号通路的影响.方法成年健康雄性Wistar大鼠,体重220～ 280 g,建立Langendorff离体心脏灌注模型.取离体心脏模型36个,采用随机数字法,将其随机分为3组(n=12):假手术组(S组)、心肌缺血再灌注组(I/R组)和七氟醚预处理组(SP组).平衡灌注30 min后,S组继续灌注150 min;I/R组缺血30 min,再灌注120 min;SP组用含2.4％七氟醚的K-H液灌注15min,然后洗脱5 min,缺血30 min,再灌注120 min.分别于平衡灌注末和再灌注30 min时,记录HR、左室舒张末压(LVEDP)、左室发展压(LVDP)、左心室内压最大上升速率(+dp/dtmax)和左心室内压最大下降速率(-dp/dtmax).再灌注期间行心律失常评分.于再灌注60 min时,每组取3个心脏,采用Western blot法测定心肌组织wnt3a、磷酸化GSK-3β(p-GSK-3β)和β-catenin的表达水平.再灌注120min时,每组取6个心脏,采用TTC染色法测定心肌梗死体积.结果与S组比较,I/R组再灌注30min时HR、LVDP、+dp/dtmax和-dp/dtmax降低,LVEDP升高,心律失常评分升高,心肌梗死体积百分比增加,心肌组织wnt3a、p-GSK-3β和β-catenin的表达下调(P＜0.05);与I/R组比较,SP组再灌注30min时HR、LVDP、+dp/dtmax和-dp/dtmax升高,LVEDP降低,心律失常评分降低,心肌梗死体积百分比减小,心肌组织wnt3a、p-GSK-3β和β-catenin的表达上调(P＜0.05).结论七氟醚预处理可通过激活wnt/GSK-3β/β-catenin信号通路减轻大鼠心肌缺血再灌注损伤.
목적 평개칠불미예처리대대서심기결혈재관주시wnt/당원합성매격매-3β(GSK-3β)/β-련배단백(β-catenin)신호통로적영향.방법 성년건강웅성Wistar대서,체중220～ 280 g,건립Langendorff리체심장관주모형.취리체심장모형36개,채용수궤수자법,장기수궤분위3조(n=12):가수술조(S조)、심기결혈재관주조(I/R조)화칠불미예처리조(SP조).평형관주30 min후,S조계속관주150 min;I/R조결혈30 min,재관주120 min;SP조용함2.4％칠불미적K-H액관주15min,연후세탈5 min,결혈30 min,재관주120 min.분별우평형관주말화재관주30 min시,기록HR、좌실서장말압(LVEDP)、좌실발전압(LVDP)、좌심실내압최대상승속솔(+dp/dtmax)화좌심실내압최대하강속솔(-dp/dtmax).재관주기간행심률실상평분.우재관주60 min시,매조취3개심장,채용Western blot법측정심기조직wnt3a、린산화GSK-3β(p-GSK-3β)화β-catenin적표체수평.재관주120min시,매조취6개심장,채용TTC염색법측정심기경사체적.결과 여S조비교,I/R조재관주30min시HR、LVDP、+dp/dtmax화-dp/dtmax강저,LVEDP승고,심률실상평분승고,심기경사체적백분비증가,심기조직wnt3a、p-GSK-3β화β-catenin적표체하조(P＜0.05);여I/R조비교,SP조재관주30min시HR、LVDP、+dp/dtmax화-dp/dtmax승고,LVEDP강저,심률실상평분강저,심기경사체적백분비감소,심기조직wnt3a、p-GSK-3β화β-catenin적표체상조(P＜0.05).결론 칠불미예처리가통과격활wnt/GSK-3β/β-catenin신호통로감경대서심기결혈재관주손상.
Objective To evaluate the effects of sevoflurane preconditioning on wnt/glycogen synthase kinase-3 beta (GSK-3β)/β-catenin signaling pathway during myocardial ischemia-reperfusion (I/R) injury in rats in vitro.Methods Ault male Wistar rats,weighing 220-280 g,were heparinized and anesthetized with intraperitoneal 3％ pentobarbital 30 mg/kg.Their hearts were rapidly excised and perfused in a langendorff apparatus with oxygenated (95％ O2-5％ CO2) K-H solution at 37 ℃.After 15 min of equilibration,36 isolated hearts were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and sevoflurane preconditioning group (group SP).After 30 min of equilibration,the hearts were continuously perfused for 150 min in group S.The isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion.In SP group,the hearts were perfused for 15 min with K-H solution containing 2.4％ sevoflurane,followed by 5 min washout before reperfusion.At the end of equilibration and 30 min of reperfusion,HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and ± dp/dtmax were recorded.The severity of arrhythmias was assessed during reperfusion.At 60 min of reperfusion,3 hearts in each group were chosen for measurement of expression of wnt3a,phosphor-GSK-3β (p-GSK-3β) and β-catenin (by Western blot).At 120 min of reperfusion,6 hearts in each group were chosen for determination of myocardial infarct size by TTC staining.Results Compared with group S,HR,LVDP,+dp/dtmax and -dp/dtmax were significantly decreased,and LVEDP was increased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were increased,and the expression of wnt3a,p-GSK-3β and β-catenin was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+dp/dtmax and-dp/dtmax were significantly increased,and LVEDP was decreased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were decreased,and the expression of wnt3a,p-GSK-3β and β-catenin was up-regulated in group SP.Conclusion Sevoflurane preconditioning attenuates myocardial I/R injury by activating wnt/GSK-3β/β-catenin signaling pathway in isolated rat hearts.