AKT通路在大鼠弥漫性轴索损伤中的作用  

Role of AKT signal pathway in diffuse axonal injury of rats

作　　者：金涛[1] 李东波 杨涛[1] 李从进[1] 宋锦宁[3] 王松林[1] JIN Tao;LI Dongbo;YANG Tao;LI Congjin;SONG Jinning;WANG Songlin(Department of Neurosurgery,Ankang City Central Hospital,Ankang 725000,China;Department of Neurosurgery,Ankang Hospital of Traditional Chinese Medicine;Department of Neurosurgery,First Affiliated Hospital of Xi’an Jiaotong University)

机构地区：[1]陕西省安康市中心医院神经外科,安康725000 [2]陕西省安康市中医院神经外科 [3]西安交通大学第一附属医院神经外科

出　　处：《山西医科大学学报》2021年第5期632-637,共6页Journal of Shanxi Medical University

基　　金：教育部新世纪优秀人才支持计划资助项目(NCET-05-0831)。

摘　　要：目的研究AKT信号通路在弥漫性轴索损伤(diffuse axonal injury,DAI)中的作用。方法雄性SD大鼠48只按随机数字表法平均分为对照组、DAI 1 d组、DAI 1 d+溶剂(vehicle)组、DAI 1 d+LY294002(LY)组。对照组为正常大鼠,仅接受麻醉;利用瞬间旋转损伤装置建立大鼠DAI模型,DAI 1 d+vehicle组在造模后即刻侧脑室注射二甲基亚砜(DMSO),DAI 1 d+LY组在造模后即刻侧脑室注射LY294002(AKT通路抑制剂,以DMSO溶解)。利用免疫组化检测神经微丝蛋白-H(NF-H)的表达;利用Western blot检测cleaved Caspase-3(C-Caspase-3)、Caspase-3、Occludin-1、闭锁小带蛋白1(ZO-1)、Claudin-5、基质金属蛋白酶9(MMP-9)、β淀粉样前体蛋白(β-APP)、Tau46、胶质纤维酸性蛋白(GFAG)、Iba-1的表达;利用Western blot检测蛋白激酶B(AKT)及GSK-3β的磷酸化水平。结果与对照组相比,DAI 1 d组NF-H的表达明显升高(P<0.05);DAI 1 d组、DAI 1 d+vehicle组、DAI 1 d+LY组的Caspase-3、C-Caspase-3、MMP-9、β-APP、GFAP、Iba-1的表达升高(均P<0.05),Occludin-1、Claudin-5、ZO-1、Tau46的表达减少(均P<0.05),p-AKT及p-GSK-3β的水平降低(均P<0.05)。与DAI 1 d组相比,DAI 1 d+vehicle组Caspase-3、C-Caspase-3、MMP-9、β-APP、GFAP、Iba-1、Occludin-1、Claudin-5、ZO-1、Tau46、p-AKT及p-GSK-3β的表达均无明显差异(均P>0.05);DAI 1 d+LY组的Caspase-3、C-Caspase-3、MMP-9、β-APP、GFAP、Iba-1的表达量更高(均P<0.05),Occludin-1、Claudin-5、ZO-1、Tau46的表达量更少(均P<0.05),p-AKT、p-GSK-3β的水平更低(均P<0.05)。结论DAI后AKT通路被抑制,其通过调控下游GSK-3β的磷酸化水平破坏血脑屏障,加重轴索损伤和细胞凋亡。Objective To study the role of AKT pathway in diffuse axonal injury(DAI)in rats.Methods Forty-eight SD male rats were randomly divided into control group,DAI 1 d group,DAI 1 d+vehicle group and DAI 1 d+LY294002(LY)group according to the random number table method.Normal rats in control group were only anesthetized.DAI model was established using a lateral head-rotation device.Rats in DAI 1 d+vehicle group were injected with DMSO to lateral cerebral ventricle immediately after injury,while rats in DAI 1 d+LY group were injected with LY294002(inhibitor of AKT pathway,dissolved in DMSO)immediately after injury.The expression of neurofilament-H(NF-H)was detected by immunohistochemistry.The expression of cleaved Caspase-3(C-Caspase-3),Caspase-3,Occludin-1,zonula occludens 1(ZO-1),Claudin-5,matrix metalloprotein 9(MMP-9),β-amyloid precursor protein(β-APP),Tau46,glial fibrillary acidic protein(GFAP),Iba-1 and the phosphorylation levels of ser-ine-threonine protein kinase(AKT)and GSK-3βwere detected by Western blot.Results Compared to control group,the expression of NF-H was increased in DAI 1 d group(P<0.05).Compared to control group,the expression levels of Caspase-3,C-Caspase-3,MMP-9,β-APP,GFAP,Iba-1 were increased,while the expression levels of Occludin-1,Claudin-5,ZO-1,Tau46 were reduced(all P<0.05),and the phosphorylation levels of AKT and GSK-3βwere decreased(all P<0.05).Compared to DAI 1 d group,the expression levels of Caspase-3,C-Caspase-3,MMP-9,β-APP,GFAP,Iba-1,Occludin-1,Claudin-5,ZO-1,Tau46,p-AKT and p-GSK-3βshowed no significant changes in DAI 1 d+vehicle group(all P>0.05).Compared with DAI 1 d group,the expression levels of Caspase-3,C-Caspase-3,MMP-9,β-APP,GFAP,Iba-1 were increased in DAI 1 d+LY group,while the expression lvels of Occludin-1,Claudin-5,ZO-1,Tau46 were reduced(all P<0.05),and the phosphorylation levels of AKT,GSK-3βwere decreased in DAI 1 d+LY group(all P<0.05).Conclusion AKT pathway can be inhibited after DAI and the inhibition could disrupt blood brain barrier,aggravate axonal in

关 键 词：AKT信号通路 凋亡 胶质反应 血脑屏障 弥漫性轴索损伤 

分 类 号：R363[医药卫生—病理学]