CAJ | 학술논문

目的探讨急性等容量血液稀释(ANH)对大鼠脑缺血再灌注时海马组织细胞凋亡的影响.方法健康清洁级雄性SD大鼠36只,体重280 ～ 320 g,50～60 d龄,采用随机数字表法,将其随机分为3组(n=12),假手术组(S组)仅游离暴露双侧椎动脉及颈总动脉；脑缺血再灌注组(I/R组)和ANH组采用四血管阻断法制备全脑缺血再灌注损伤模型.ANH组于灼烧双侧椎动脉后24h实施ANH,经股动脉放血(经10 min),放血的同时股静脉输注6％羟乙基淀粉130/0.4氯化钠,ANH结束后10 min时夹闭双侧颈总动脉5 min.于再灌注24 h时,处死大鼠,取脑组织,分离海马,采用流式细胞术测定细胞凋亡情况,采用反转录聚合酶链反应法测定Apaf-1 mRNA及caspase-3 mRNA的表达.结果与S组比较,I/R组和ANH组海马组织细胞凋亡率升高,海马组织Apaf-1 mRNA及caspase-3mRNA表达上调(P＜0.01)；与I/R组比较,ANH组海马组织细胞凋亡率降低,海马组织Apaf-1 mRNA及caspase-3 mRNA表达下调(P＜0.01).结论 ANH可抑制大鼠全脑缺血再灌注时海马神经细胞凋亡,其机制与下调Apaf-1及caspase-3表达有关.
목적 탐토급성등용량혈액희석(ANH)대대서뇌결혈재관주시해마조직세포조망적영향.방법 건강청길급웅성SD대서36지,체중280 ～ 320 g,50～60 d령,채용수궤수자표법,장기수궤분위3조(n=12),가수술조(S조)부유리폭로쌍측추동맥급경총동맥；뇌결혈재관주조(I/R조)화ANH조채용사혈관조단법제비전뇌결혈재관주손상모형.ANH조우작소쌍측추동맥후24h실시ANH,경고동맥방혈(경10 min),방혈적동시고정맥수주6％간을기정분130/0.4록화납,ANH결속후10 min시협폐쌍측경총동맥5 min.우재관주24 h시,처사대서,취뇌조직,분리해마,채용류식세포술측정세포조망정황,채용반전록취합매련반응법측정Apaf-1 mRNA급caspase-3 mRNA적표체.결과 여S조비교,I/R조화ANH조해마조직세포조망솔승고,해마조직Apaf-1 mRNA급caspase-3mRNA표체상조(P＜0.01)；여I/R조비교,ANH조해마조직세포조망솔강저,해마조직Apaf-1 mRNA급caspase-3 mRNA표체하조(P＜0.01).결론 ANH가억제대서전뇌결혈재관주시해마신경세포조망,기궤제여하조Apaf-1급caspase-3표체유관.
Objective To investigate the effect of acute normovolemic hemodilution (ANH) on the apoptosis in hippocampal cells induced by global cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy 50-60 day old male SD rats weighing 280-320 g were randomly divided into 3 groups ( n =12 each):group sham operation (group S); group global cerebral I/R (group I/R) and group ANH.Global cerebral I/R was produced by 4-vessel technique described by Pulsinelli et al.in groups I/R and ANH.ANH was carried out at 24 h after cauterization of bilateral vertebral arteries,before occlusion of bilateral carotid arteries.Blood was withdrawn from femoral artery until Hct was reduced to 30％ and equal volume of hydroxyethyl starch 130/0.4 sodium chloride was infused into femoral vein simultaneously.Bilateral carotid arteries were blocked for 5 min at 10 min after ANH.The rats were sacrificed at 24 h of reperfusion and their hippocampi were isolated.Apoptosis was detected by flow cytometry.The expression of Apaf-1 mRNA and caspase-3 mRNA was determined by RT-PCR.Results Global cerebral I/R significantly increased apoptosis index and up-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group I/R as compared with group S.ANH significantly attenuated apoptosis and down-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group ANH compared with group I/R.Conclusion ANH can reduce hippocampal cell apoptosis induced by cerebral I/R through down-regulation of Apaf-1 and caspase-3 expression in hippocampus.