CAJ | 학술논문

目的探讨反复双侧颈总动脉闭塞所致血管性痴呆(vascular dementia,VaD)小鼠海马Akt和口糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK3β)蛋白表达.方法 48只健康雄性C57B1/6小鼠随机分为正常组、假手术组和模型组,每组16只.模型组采用反复3次间断阻断双侧颈总动脉建立小鼠VaD模型,假手术组只分离双侧颈总动脉但不阻断,正常组不接受任何处理.术后4周利用水迷宫实验和跳台实验观测各组小鼠行为学改变,采用HE染色观察海马组织病理学变化,采用蛋白质印迹法检测海马Akt、p-Akt(Ser473)、GSK3β、p-GSK3β(Ser9)蛋白表达.结果水迷宫实验中,模型组学习阶段和记忆阶段游完全程时间延长(学习阶段:F=19.389,P＜0.05;记忆阶段:F=27.929,P＜0.05),错误次数增加(学习阶段:F=7.228,P＜0.05;记忆阶段:F =21.189,P＜0.05)；跳台实验中,模型组小鼠学习阶段反应时间延长(F=19.162,P＜0.05)、错误次数增加(F=6.562,P＜0.05),记忆阶段潜伏时间缩短(F=10.634,P＜0.05)、错误次数增加(F=12.890,P＜0.05).同时,HE染色显示模型组海马CA1区神经元减少,神经胶质细胞增生；p-Akt(Ser473)(F=37.849,P＜0.05)和p-GSK3β(Ser9)(F=67.725,P＜0.05)蛋白表达在术后4周较假手术组显著上调,而各组间Akt(F=1.004,P＞0.05)和GSK3β(F=0.329,P＞0.05)总蛋白表达无显著差异.结论反复双侧颈总动脉闭塞可造成小鼠学习记忆障碍,海马组织受损严重.磷酸化Akt和GSK3β表达可能参与了VaD的机制.
목적 탐토반복쌍측경총동맥폐새소치혈관성치태(vascular dementia,VaD)소서해마Akt화구당원합성매격매-3β(glycogen synthase kinase-3β,GSK3β)단백표체.방법 48지건강웅성C57B1/6소서수궤분위정상조、가수술조화모형조,매조16지.모형조채용반복3차간단조단쌍측경총동맥건립소서VaD모형,가수술조지분리쌍측경총동맥단불조단,정상조불접수임하처리.술후4주이용수미궁실험화도태실험관측각조소서행위학개변,채용HE염색관찰해마조직병이학변화,채용단백질인적법검측해마Akt、p-Akt(Ser473)、GSK3β、p-GSK3β(Ser9)단백표체.결과 수미궁실험중,모형조학습계단화기억계단유완전정시간연장(학습계단:F=19.389,P＜0.05;기억계단:F=27.929,P＜0.05),착오차수증가(학습계단:F=7.228,P＜0.05;기억계단:F =21.189,P＜0.05)；도태실험중,모형조소서학습계단반응시간연장(F=19.162,P＜0.05)、착오차수증가(F=6.562,P＜0.05),기억계단잠복시간축단(F=10.634,P＜0.05)、착오차수증가(F=12.890,P＜0.05).동시,HE염색현시모형조해마CA1구신경원감소,신경효질세포증생；p-Akt(Ser473)(F=37.849,P＜0.05)화p-GSK3β(Ser9)(F=67.725,P＜0.05)단백표체재술후4주교가수술조현저상조,이각조간Akt(F=1.004,P＞0.05)화GSK3β(F=0.329,P＞0.05)총단백표체무현저차이.결론 반복쌍측경총동맥폐새가조성소서학습기억장애,해마조직수손엄중.린산화Akt화GSK3β표체가능삼여료VaD적궤제.
Objective To investigate the expressions of protein kinase B (PKB/Akt) and glycogen synthase kinasc-3β in the hippocampus in mice with vascular dementia (VaD) induced by repetitive bilateral common carotid artery occlusion.Methods Forty-eight healthy adult male C57B1/6 mice were randomly allocated into 3 group:normal group,sham operation group,and model group (n =16 in each group).A mouse VaD model was induced by intermittent blocking the bilateral common carotid artery for 3 times in the model group.The sham group only separated the bilateral common carotid artery,but did not block it.The normal group did not receive any treatment.The behavioral changes of the mice were observed using the water maze and step-down tests at 4 weeks after procedure.HE staining was used to observe the histopathological changes of hippocampal tissue.The Western blotting was used to detect the expressions of Akt,p-Akt (Ser473),GSK3β and p-GSK3β (Ser9) proteins.Results In the water maze test,the time of swimming the entire distance was prolonged at the learning stage and memory stage (learning stage:F =19.389,P ＜0.05; memory stage:F =27.929,P ＜ 0.05),the number of errors increased (learning stage:F =7.228,P ＜ 0.05; memory stage:F =21.189,P＜0.05) in the model group.In the step-down test,the response time was prolonged (F=19.162,P ＜0.05) at learning stage and the number of errors increased (F =6.562,P ＜ 0.05),the latency time was shortened (F=10.634,P＜0.05) and the number of errors increased (F=12.890,P＜0.05) in the model group.At the same time,HE staining showed the reduction of neurons and the proliferation of glial cells in the hippocampal CA1 region in the model group; p-Akt (Ser473) (F=37.849,P＜0.05) and p-GSK3β (Ser9)(F =67.725,P ＜0.05) protein expressions were up-regulated significantly (F =37.849,P ＜0.05; F =67.725,P＜0.05) at 4 weeks after procedure compared to those in the sham operation group,while there were no significant differences in Akt (F =1.004,P ＞0.05) and GSK3β(F =0.329,P ＞0.05) total protein expressions among all groups.Conclusions The repetitive bilateral common carotid artery occlusion may result in learning and memory impairment and severe damage in the hippocampus in mice.The Akt and GSK3β expressions may be involved in the mechanism of VaD.