CAJ | 학술논문

目的探讨抑制AMP-活化蛋白激酶(AMP-activated protein kinase,AMPK)活性对小鼠脑缺血再灌注后胱天蛋白酶(caspase)-3表达和神经细胞凋亡的影响.方法 36只雄性C57BL/6小鼠,按随机数字表法分为假手术组、缺血再灌注组和AMPK抑制剂组,每组12只.线栓法建立小鼠大脑中动脉闭塞模型,AMPK抑制剂组在插入线栓时腹腔注射AMPK抑制剂Compound C(20 mg/kg),假手术组和缺血再灌注组在相同时间点腹腔注射等体积生理盐水.脑缺血再灌注24 h后应用免疫组化染色法检测胱天蛋白酶-3表达水平,应用TUNEL法检测神经细胞凋亡.结果 AMPK抑制剂组皮质胱天蛋白酶-3阳性细胞[(7.16±5.85)个/高倍视野对(14.36±7.85)个/高倍视野;t=2.548,P=0.018]和TUNEL阳性细胞[(58.86±9.65)个/高倍视野对(81.00±12.21)个/高倍视野;拇4.928,P＜0.001]数量均显著少于缺血再灌注组.结论抑制AMPK活性可下调脑缺血再灌注后缺血皮质胱天蛋白酶-3表达,减少神经细胞凋亡.
목적 탐토억제AMP-활화단백격매(AMP-activated protein kinase,AMPK)활성대소서뇌결혈재관주후광천단백매(caspase)-3표체화신경세포조망적영향.방법 36지웅성C57BL/6소서,안수궤수자표법분위가수술조、결혈재관주조화AMPK억제제조,매조12지.선전법건립소서대뇌중동맥폐새모형,AMPK억제제조재삽입선전시복강주사AMPK억제제Compound C(20 mg/kg),가수술조화결혈재관주조재상동시간점복강주사등체적생리염수.뇌결혈재관주24 h후응용면역조화염색법검측광천단백매-3표체수평,응용TUNEL법검측신경세포조망.결과 AMPK억제제조피질광천단백매-3양성세포[(7.16±5.85)개/고배시야대(14.36±7.85)개/고배시야;t=2.548,P=0.018]화TUNEL양성세포[(58.86±9.65)개/고배시야대(81.00±12.21)개/고배시야;무4.928,P＜0.001]수량균현저소우결혈재관주조.결론 억제AMPK활성가하조뇌결혈재관주후결혈피질광천단백매-3표체,감소신경세포조망.
Objective To investigate the effects of inhibition of adenosine monophosphate-activated protein kinase (AMPK) activity on caspase-3 expression and neuronal apoptosis after cerebral ischemiareperfusion in mice.Methods A total of 36 male C57BL/6 mice were randomly divided into a sham operation group,an ischemia-reperfusion group and an AMPK inhibitor group (n =12 in each group).A model of middle cerebral artery occlusion (MCAO) was induced by the suture method.AMPK inhitor Compound C (20 mg/kg) was injected intraperitoneally whie inserting sutures in the AMPK inhibitor group.The equivalent volumes of normal saline solution was injected intraperitoneally at the same tine points in the sham operation group and the ischemia-reperfusion group.At 24 h after cerebral ischemia-reperfusion,immunohistochemical staining was use to detect the expression levels of caspase-3.TUNEL method was used to detect neuronal apoptosis in mice.Results Compared with the ischemia-reperfusion group,the numbers of the cortical caspase-3 positive cells (7.16 ±5.85 vs.14.36 ±7.85;t =2.548,P =0.018) and the TUNEL positive cells (58.86 ±9.65 vs.81.00 ± 12.21;t =4.928,P ＜0.001) per high-power field in the AMPK inhibitor group were decreased significantly.Conclmions Inhibition of AMPK activity downregulates the expression ofcaspase-3 after cerebral ischemia-reperfusion,and reduces neuronal apoptosis.