CAJ | 학술논문

目的：评价磷酸腺苷活化蛋白激酶（ AMPK）在老龄小鼠脑缺血再灌注时海马神经元凋亡中的作用。方法健康清洁级雄性C57∕BL6小鼠72只，18～20月龄，体重34～36 g，采用随机数字表法分为3组（ n＝24）：假手术组（ S组）、脑缺血再灌注组（ I∕R组）和AMPK抑制剂Compound C组（ CC组）。采用夹闭双侧颈总动脉15 min后再灌注的方法制备脑缺血再灌注损伤模型。 CC组于夹闭双侧颈总动脉即刻腹腔注射Compound C 20 mg∕kg，I∕R组于相同时点给予等容量生理盐水。于再灌注48 h时处死，取海马组织，HE染色观察海马CA1区神经元病理学结果；采用TUNEL法检测神经元凋亡率；采用Western blot法检测海马磷酸化AMPKα（ p?AMPKα）和caspase?3的表达。结果I∕R组和CC组海马CA1区神经元发生病理学损伤，锥体细胞排列紊乱，细胞核固缩浓染。与S组比较，I∕R组和CC组再灌注48 h时神经元凋亡率升高，p?AMPKα及caspase?3表达上调（ P＜0．05）。与I∕R组比较，CC组海马p?AMPKα表达下调（ P＜0．05），神经元凋亡率和caspase?3表达差异无统计学意义（ P＞0．05）。结论 AMPK与老龄小鼠脑缺血再灌注时海马神经元凋亡无明显关系。
목적：평개린산선감활화단백격매（ AMPK）재노령소서뇌결혈재관주시해마신경원조망중적작용。방법건강청길급웅성C57∕BL6소서72지，18～20월령，체중34～36 g，채용수궤수자표법분위3조（ n＝24）：가수술조（ S조）、뇌결혈재관주조（ I∕R조）화AMPK억제제Compound C조（ CC조）。채용협폐쌍측경총동맥15 min후재관주적방법제비뇌결혈재관주손상모형。 CC조우협폐쌍측경총동맥즉각복강주사Compound C 20 mg∕kg，I∕R조우상동시점급여등용량생리염수。우재관주48 h시처사，취해마조직，HE염색관찰해마CA1구신경원병이학결과；채용TUNEL법검측신경원조망솔；채용Western blot법검측해마린산화AMPKα（ p?AMPKα）화caspase?3적표체。결과I∕R조화CC조해마CA1구신경원발생병이학손상，추체세포배렬문란，세포핵고축농염。여S조비교，I∕R조화CC조재관주48 h시신경원조망솔승고，p?AMPKα급caspase?3표체상조（ P＜0．05）。여I∕R조비교，CC조해마p?AMPKα표체하조（ P＜0．05），신경원조망솔화caspase?3표체차이무통계학의의（ P＞0．05）。결론 AMPK여노령소서뇌결혈재관주시해마신경원조망무명현관계。
Objective To evaluate the role of AMP?activated protein kinase ( AMPK) in neuronal apoptosis in the hippocampus of aged mice during cerebral ischemia?reperfusion ( I∕R ) . Methods Seventy?two male C57∕BL6 mice, aged 18-20 months, weighing 34-36 g, were randomly divided into 3 groups ( n=24 each ) using a random number table: sham operation group ( S group ) , I∕R group and AMPK inhibitor compound C group ( CC group) . The cerebral ischemia was produced by 15 min transient occlusion of bilateral common carotid arteries followed by reperfusion. In group CC, compound C 20 mg∕kg was injected intraperitoneally immediately after occlusion, while the equal volume of normal saline was given in I∕R group. At 48 h of reperfusion, the mice were sacrificed, and the brains were immediately harvested for examination of pathologic changes in hippocampal CA1 region and for determination of neuronal apoptosis ( using TUNEL) and expression of phosphor?AMPKα ( p?AMPKα) and caspase?3 ( by Western blot) . Results In I∕R and CC groups, the examination showed the pathologic changes in hippocampal CA1 region, the disordered arrangement of pyramidal cells and the nucleus condensation. Compared with S group, the neuronal apoptotic rate was significantly increased, and the expression of p?AMPK and caspase?3 was up?regulated at 48 h of reperfusion in I∕R and CC groups. Compared with I∕R group, the expression of p?AMPK was significantly down?regulated, and no significant change was found in the neuronal apoptotic rate and caspase?3 expression in CC group. Conclusion AMPK is not involved in neuronal apoptosis in the hippocampus of aged mice during cerebral I∕R.