国际脑血管病杂志
國際腦血管病雜誌
국제뇌혈관병잡지
INTERNATIONAL JOURNAL OF CEREBROVASCULAR DISEASES
2013年
8期
624-629
,共6页
何敏%韩江全%倪白云%张险平
何敏%韓江全%倪白雲%張險平
하민%한강전%예백운%장험평
脑缺血%蛋白质丝氨酸苏氨酸激酶%磷脂酰肌醇3-激酶类%bcl-死亡相关蛋白%原癌基因蛋白质c-akt%细胞凋亡%疾病模型,动物%大鼠
腦缺血%蛋白質絲氨痠囌氨痠激酶%燐脂酰肌醇3-激酶類%bcl-死亡相關蛋白%原癌基因蛋白質c-akt%細胞凋亡%疾病模型,動物%大鼠
뇌결혈%단백질사안산소안산격매%린지선기순3-격매류%bcl-사망상관단백%원암기인단백질c-akt%세포조망%질병모형,동물%대서
Brain Ischemia%Protein-Serine-Threonine Kinases%Phosphatidy linositol 3-Kinases%bcl-Associated Death Protein%Proto-Oncogene Proteins c-akt%Disease Models,Animal%Apoptosis%Rats
目的 探讨永久性大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠脑梗死周围组织p-Akt(Ser473)、p-Bad(Serl36)表达的变化及其与细胞凋亡关系.方法 60只雄性Sprague-Dawley大鼠随机分为假手术、MCAO 3 h、MCAO 12 h、LY294002干预MCAO 3 h和 LY294002干预MCAO 12 h组,每组12只.改良线栓法制作永久性MCAO模型,LY294002干预组在模型制作前15min经侧脑室给药.采用Longa法进行神经功能评分,氯化三苯基四氮唑(2,3,5-triphenyl tetrazolium chloride,TTC)法检测梗死体积,免疫组织化学染色法检测梗死周围组织p-Akt(Ser473)和p-Bad (Ser136)表达,TUNEL法检测梗死周围组织凋亡细胞.结果 模型制作后3h,所有实验大鼠均从麻醉中清醒.假手术组神经功能缺损评分为0分,MCAO 3 h组、MCAO 12 h组、LY294002干预MCAO 3 h组和LY294002干预MCAO 12 h组分别为(2.25±0.45)、(2.92±0.99)、(3.00±0.95)和(3.02 ±0.36)分,各组间存在显著性差异(F =26.520,P=0.000).其中,LY294002干预MCAO 3 h组显著性高于MCAO 3 h组(P=0.009),LY294002干预MCAO 12 h组与MCAO 12 h组无显著性差异(P=0.354).TTC染色显示,假手术组未见梗死灶,MCAO 3 h组、MCAO 12 h组、LY294002干预MCAO 3 h组和LY294002干预MCAO 12 h组梗死体积分别为(23.4±1.4)、(40.3±1.1)、(31.9±6.0)和(44.4±3.8)mm3,各组间存在显著性差异(F=30.440,P=0.000).其中,LY294002干预MCAO 3 h组显著性大于MCAO 3 h组(P =0.002),LY294002干预MCAO 12 h组与MCAO 12 h组无显著性差异(P=0.113).与假手术组相比,MCAO 3 h组梗死周围组织p-Akt(Ser473)表达显著性增高,MCAO 12 h组显著性下降；p-Bad(Ser136)表达水平随着MCAO时间的推移呈现进行性下降,同时凋亡细胞数量进行性增多.LY294002干预后,MCAO 3 h时梗死周围组织p-Akt(Ser473)和p-Bad(Ser136)表达水平均显著性下降,凋亡细胞数量显著性增多(P＜0.05),但对MCAO后12 h时的各指标无显著性影响.结论 脑梗死早期P13K/Akt信号转导通路的激活以及该通路关键蛋白p-Akt(Ser473)的应激性升高具有脑保护作用,而脑梗死后期该通路活性的衰竭及其关键蛋白的急剧降低则与脑损伤有关.
目的 探討永久性大腦中動脈閉塞(middle cerebral artery occlusion,MCAO)大鼠腦梗死週圍組織p-Akt(Ser473)、p-Bad(Serl36)錶達的變化及其與細胞凋亡關繫.方法 60隻雄性Sprague-Dawley大鼠隨機分為假手術、MCAO 3 h、MCAO 12 h、LY294002榦預MCAO 3 h和 LY294002榦預MCAO 12 h組,每組12隻.改良線栓法製作永久性MCAO模型,LY294002榦預組在模型製作前15min經側腦室給藥.採用Longa法進行神經功能評分,氯化三苯基四氮唑(2,3,5-triphenyl tetrazolium chloride,TTC)法檢測梗死體積,免疫組織化學染色法檢測梗死週圍組織p-Akt(Ser473)和p-Bad (Ser136)錶達,TUNEL法檢測梗死週圍組織凋亡細胞.結果 模型製作後3h,所有實驗大鼠均從痳醉中清醒.假手術組神經功能缺損評分為0分,MCAO 3 h組、MCAO 12 h組、LY294002榦預MCAO 3 h組和LY294002榦預MCAO 12 h組分彆為(2.25±0.45)、(2.92±0.99)、(3.00±0.95)和(3.02 ±0.36)分,各組間存在顯著性差異(F =26.520,P=0.000).其中,LY294002榦預MCAO 3 h組顯著性高于MCAO 3 h組(P=0.009),LY294002榦預MCAO 12 h組與MCAO 12 h組無顯著性差異(P=0.354).TTC染色顯示,假手術組未見梗死竈,MCAO 3 h組、MCAO 12 h組、LY294002榦預MCAO 3 h組和LY294002榦預MCAO 12 h組梗死體積分彆為(23.4±1.4)、(40.3±1.1)、(31.9±6.0)和(44.4±3.8)mm3,各組間存在顯著性差異(F=30.440,P=0.000).其中,LY294002榦預MCAO 3 h組顯著性大于MCAO 3 h組(P =0.002),LY294002榦預MCAO 12 h組與MCAO 12 h組無顯著性差異(P=0.113).與假手術組相比,MCAO 3 h組梗死週圍組織p-Akt(Ser473)錶達顯著性增高,MCAO 12 h組顯著性下降；p-Bad(Ser136)錶達水平隨著MCAO時間的推移呈現進行性下降,同時凋亡細胞數量進行性增多.LY294002榦預後,MCAO 3 h時梗死週圍組織p-Akt(Ser473)和p-Bad(Ser136)錶達水平均顯著性下降,凋亡細胞數量顯著性增多(P＜0.05),但對MCAO後12 h時的各指標無顯著性影響.結論 腦梗死早期P13K/Akt信號轉導通路的激活以及該通路關鍵蛋白p-Akt(Ser473)的應激性升高具有腦保護作用,而腦梗死後期該通路活性的衰竭及其關鍵蛋白的急劇降低則與腦損傷有關.
목적 탐토영구성대뇌중동맥폐새(middle cerebral artery occlusion,MCAO)대서뇌경사주위조직p-Akt(Ser473)、p-Bad(Serl36)표체적변화급기여세포조망관계.방법 60지웅성Sprague-Dawley대서수궤분위가수술、MCAO 3 h、MCAO 12 h、LY294002간예MCAO 3 h화 LY294002간예MCAO 12 h조,매조12지.개량선전법제작영구성MCAO모형,LY294002간예조재모형제작전15min경측뇌실급약.채용Longa법진행신경공능평분,록화삼분기사담서(2,3,5-triphenyl tetrazolium chloride,TTC)법검측경사체적,면역조직화학염색법검측경사주위조직p-Akt(Ser473)화p-Bad (Ser136)표체,TUNEL법검측경사주위조직조망세포.결과 모형제작후3h,소유실험대서균종마취중청성.가수술조신경공능결손평분위0분,MCAO 3 h조、MCAO 12 h조、LY294002간예MCAO 3 h조화LY294002간예MCAO 12 h조분별위(2.25±0.45)、(2.92±0.99)、(3.00±0.95)화(3.02 ±0.36)분,각조간존재현저성차이(F =26.520,P=0.000).기중,LY294002간예MCAO 3 h조현저성고우MCAO 3 h조(P=0.009),LY294002간예MCAO 12 h조여MCAO 12 h조무현저성차이(P=0.354).TTC염색현시,가수술조미견경사조,MCAO 3 h조、MCAO 12 h조、LY294002간예MCAO 3 h조화LY294002간예MCAO 12 h조경사체적분별위(23.4±1.4)、(40.3±1.1)、(31.9±6.0)화(44.4±3.8)mm3,각조간존재현저성차이(F=30.440,P=0.000).기중,LY294002간예MCAO 3 h조현저성대우MCAO 3 h조(P =0.002),LY294002간예MCAO 12 h조여MCAO 12 h조무현저성차이(P=0.113).여가수술조상비,MCAO 3 h조경사주위조직p-Akt(Ser473)표체현저성증고,MCAO 12 h조현저성하강；p-Bad(Ser136)표체수평수착MCAO시간적추이정현진행성하강,동시조망세포수량진행성증다.LY294002간예후,MCAO 3 h시경사주위조직p-Akt(Ser473)화p-Bad(Ser136)표체수평균현저성하강,조망세포수량현저성증다(P＜0.05),단대MCAO후12 h시적각지표무현저성영향.결론 뇌경사조기P13K/Akt신호전도통로적격활이급해통로관건단백p-Akt(Ser473)적응격성승고구유뇌보호작용,이뇌경사후기해통로활성적쇠갈급기관건단백적급극강저칙여뇌손상유관.
Objective To investigate the relationship between the expression changes of p-Akt (Ser473),p-Bad (Ser136) and the cell apoptosis in peri-infarction tissue of permanent middle cerebral artery occlusion (MCAO) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into sham operation,MCAO 3 h,MCAO 12 h,LY294002 intervention MCAO 3 h,and LY294002 intervention MCAO 12 h groups (n =12 in each group).A permanent MCAO model was induced by the modified suture method.At 15 minutes before modeling,the rats in the LY294002 intervention MCAO groups were administered via lateral ventricle.The neurological function score was scored by using Zea Longa method.2,3,5-triphenyltetrazolium chloride (TTC) staining was used to detect infarct volume.Immunohistochemical staining was use to detect pAkt (Ser473) and p-Bad (Ser136) expressions in peri-infaretion tissue.TUNEL assay was used to detect apoptotic cells in peri-infarction tissue.Results Three hours after modeling all the experimental rats awoke from anesthesia.The neurological deficit score in the sham operation group was 0,and the scores of the MCAO 3 h,MCAO 12 h,LY294002 intervention MCAO 3 h and LY294002 intervention MCAO 12 h groups were 2.25 ± 0.45,2.92 ± 0.99,3.00 ± 0.95,and 3.02 ± 0.36,respectively.There were significant differences among all groups (F =26.520,P =0.000).The score of the LY294002 intervention MCAO 3 h group was significantly higher than that of the MCAO 3 h group (P =0.009).There was no significant difference between the LY294002 intervention MCAO 12 h group and the MCAO 12 h group (P =0.354).TTC staining showed that no infarct was observed in the sham operation group.The infarct volumes of the MCAO 3 h,MCAO 12 h,LY294002 intervention MCAO 3 h and LY294002 intervention MCAO 12 h groups were 23.4 ± 1.4,40.3 ± 1.1,31.9 ±6.0 and 44.4 ±3.8 mm3,respectively.There were significant differences among the groups (F =30.440,P =0.000).The score of the LY294002 intervention MCAO 3 h group was significantly greater than that of the MCAO 3 h group (P =0.002).There was no significant difference between the LY294002 intervention MCAO 12 h group and the MCAO 12 h group (P=0.113).Compared with the sham operation group,the p-Akt (Ser473) expression in peri-infarction tissue in the MCAO 3 h group was significantly high,and it was significantly decreased in the MCAO 12 h group; the expression level of p-Bad (Ser136) showed a progressive decline with the passage of MCAO time,at the same time,the number of apoptotic cells increased progressively.After the LY294002 intervention,the expression levels of p-Akt (Ser473) and p-Bad (Ser136) in peri-infarction tissue decreased significantly at 3 h after MCAO,and the number of apoptotic cells increased significantly (P ＜0.05),but there was no significant effect on each index at 12 h after MCAO.Conclusions The activation of the PI3K/Akt signal transduction pathway in early cerebral infarction and the stress elevation of the key protein p-Akt (Ser473) of this pathway have brain protection,while the failure of this pathway activity and the drastical decrease of its key protein in the late cerebral infarction are associated with brain injury.
