肉苁蓉总苷经PI3K-Akt信号通路对大鼠脑缺血再灌注损伤的神经保护作用及机制研究  

Neuroprotective Effect and Mechanism of Glycosides of Cistanche on Cerebral Ischemia-Reperfusion Injury in Rats Based on PI3K-Akt Signaling Pathway

作　　者：王璐[1] 张士滨 吕雪[2] 杨占君[3] 贾建新[3] 吴丽娥[1] WANG Lu;ZHANG Shibin;LYU Xue;YANG Zhanjun;JIA Jianxin;WU Li’e(Department of Neurology,The First Affiliated Hospital of Baotou Medical College,Baotou 014040,China;Graduate School of Baotou Medical College,Baotou 014040,China;Department of Human Anatomy,Baotou Medical College,Baotou 014040,China)

机构地区：[1]包头医学院第一附属医院神经内科,包头014040 [2]包头医学院研究生学院 [3]包头医学院人体解剖学教研室

出　　处：《中国卒中杂志》2024年第11期1321-1332,共12页Chinese Journal of Stroke

基　　金：国家自然科学基金项目(81860215);内蒙古自治区自然科学基金项目(2021MS08010)。

摘　　要：目的探究肉苁蓉总苷通过激活PI3K-Akt信号通路对大鼠脑缺血再灌注损伤(cerebral ischemiareperfusion injury,CIRI)的保护作用及机制。方法60只雄性Wistar大鼠随机分为假手术组、模型组、肉苁蓉总苷组、尼莫地平组。采用改良线栓法构建大鼠CIRI模型。采用Zea-Longa神经功能评分评估各组大鼠中枢神经系统缺损状况,2,3,5-氯化三苯基四氮唑染色计算各组模型大鼠脑梗死面积,原位末端转移酶标记法染色检测细胞凋亡率,免疫荧光染色分析凋亡相关因子的表达情况,蛋白免疫印迹以及荧光定量PCR检测各组大鼠凋亡相关因子及PI3K-Akt信号通路相关分子表达水平。结果与假手术组相比,CIRI模型大鼠神经系统缺损症状严重,神经功能评分升高(P<0.05),运动能力减退,脑梗死面积增大,凋亡细胞增多,促凋亡因子B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)相关X蛋白(Bcl-2 as sociated X,Bax)和细胞色素C(cytochrome C,CytC)表达增加(P<0.05),Bcl-2、Bcl-2/Bax、磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)、磷酸化蛋白激酶B(phospho-protein kinase B,p-Akt)、p-Akt/Akt表达降低(P<0.05)。与模型组相比,肉苁蓉总苷能够促进CIRI模型大鼠运动功能的恢复,减小脑梗死面积,调控神经细胞凋亡,抑制Bax、CytC的表达(P<0.05),促进Bcl-2、Bcl-2/Bax、PI 3K、p-Akt、p-Akt/Akt的表达。结论肉苁蓉总苷可能通过激活PI3K-Akt信号通路,抑制神经细胞凋亡,对CIRI模型大鼠发挥神经保护作用。Objective To investigate the protective effects and mechanisms of glycosides of cistanche(GCs)on cerebral ischemia-reperfusion injury(CIRI)in rats by activating PI3K-Akt signaling pathway.Methods Sixty male Wistar rats were randomly divided into the sham group,the model group,the GCs group and the nimodipine group.The modified embolism method was used to construct a rat model of CIRI.The deficits in the central nervous system of rats in each group were assessed by the Zea-Longa neurological function score.The area of cerebral infarction of rats in each group was calculated by 2,3,5-triphenyltetrazolium chloride staining.The apoptosis rate was detected by terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining.The expression of apoptosis-related factors was analyzed by immunofluorescence staining.Western blot and fluorogenic quantitative PCR were used to detect the expression levels of apoptosis-related factors and molecules related to the PI3K-Akt signaling pathway in each group of rats.Results Compared with the sham group,the CIRI model rats exhibited severe neurological deficit symptoms,increased neurological function scores(P<0.05),reduced motor function,enlarged cerebral infarction area,increased apoptotic cells,and elevated expression of pro-apoptotic factors B-cell lymphoma-2(Bcl-2)associated X protein(Bax)and cytochrome C(CytC)(P<0.05).The expression levels of Bcl-2,Bcl-2/Bax,phosphoinositide 3-kinase(PI3K),phospho-protein kinase B(p-Akt),and p-Akt/Akt were decreased(P<0.05).Compared with the model group,GCs promoted the recovery of motor function,reduced the area of cerebral infarction,regulated neuronal apoptosis,inhibited the expression of Bax and CytC(P<0.05),and promoted the expression of Bcl-2,Bcl-2/Bax,PI3K,p-Akt,and p-Akt/Akt in CIRI model rats.Conclusions GCs may exert neuroprotective effects on CIRI model rats by activating the PI3K-Akt signaling pathway and inhibiting neuronal apoptosis.

关 键 词：PI3K-AKT信号通路 脑缺血再灌注 肉苁蓉总苷 细胞凋亡 神经系统 

分 类 号：R74[医药卫生—神经病学与精神病学]