基于PI3K/Akt信号通路探讨酸枣仁皂苷A改善血管性认知障碍模型大鼠认知功能的作用机制  

Jujuboside A Improves Cognitive Function in Rat Model of VCI via PI3K/Akt Signaling Pathway

作　　者：黄子轩 杨硕 周家琪 张耿超 游秋云[1,2] 谭爱华 HUANG Zixuan;YANG Shuo;ZHOU Jiaqi;ZHANG Gengchao;YOU Qiuyun;TAN Aihua(Ministry of Education Engineering Research Centre of Chinese Medicine Protection Technology and New Product Development for Elderly Brain Health,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China;Huanggang Hospital of Traditional Chinese Medicine Affiliated to Hubei University of Chinese Medicine,Huanggang 438000,China)

机构地区：[1]湖北中医药大学老年脑健康中医药防护技术与新产品研发教育部工程研究中心,武汉430065 [2]湖北时珍实验室,武汉430065 [3]湖北中医药大学附属黄冈中医医院,湖北黄冈438000

出　　处：《中国实验方剂学杂志》2025年第6期107-114,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(82405523);湖北省自然科学基金项目(2022CFB962,2024AFB922);湖北省中医药管理局中医药青年人才项目(ZY2023Q025)。

摘　　要：目的:观察酸枣仁皂苷A(JuA)对血管性认知障碍(VCI)大鼠学习记忆的影响及组织病理变化,探索酸枣仁皂苷A对于治疗血管性认知障碍的作用机制。方法:SPF级雄性SD大鼠50只,从中各随机选择出10只作为假手术组和正常组,其余大鼠进行双侧颈总动脉结扎法(2-VO)建立VCI模型。将手术后情况稳定的大鼠随机平均分为模型组、酸枣仁皂苷A组(20 mg·kg^(-1))、多奈哌齐组(0.45 mg·kg^(-1))。连续灌胃4周后通过新物体识别实验和Morris水迷宫实验评价酸枣仁皂苷A对VCI模型大鼠学习记忆能力的影响;尼氏染色评价海马神经元阳性细胞数目和形态变化;实时荧光定量聚合酶链式反应(Realtime PCR)检测大鼠海马组织糖原合成酶激酶-3β(GSK-3β)、cAMP反应元件结合蛋白(CREB)、B细胞淋巴瘤-2(Bcl-2)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)mRNA相对表达量。蛋白免疫印迹法(Western blot)检测大鼠海马组织GSK-3β、磷酸化(p)-GSK-3β、p-CREB、Bcl-2、PI3K、p-PI3K、Akt、p-Akt蛋白的表达。结果:与假手术组比较,模型组大鼠学习记忆能力显著下降(P<0.01),海马CA1区神经元细胞损伤,细胞数量减少(P<0.01),海马组织中GSK-3βmRNA表达水平显著升高(P<0.01),PI3K、Akt、CREB、Bcl-2 mRNA表达水平显著降低(P<0.01),p-PI3K、p-Akt、p-GSK-3β、p-CREB、Bcl-2的蛋白表达水平显著降低(P<0.01);与模型组比较,JuA组和多奈哌齐组大鼠学习记忆能力明显提高(P<0.05,P<0.01),海马CA1区神经元细胞损伤程度降低,细胞数量增加(P<0.05,P<0.01),海马组织中GSK-3βmRNA表达水平显著降低(P<0.01),PI3K、Akt、CREB、Bcl-2的mRNA表达水平明显升高(P<0.05,P<0.01),海马组织中p-PI3K、p-Akt、p-GSK-3β、p-CREB、Bcl-2的蛋白表达水平明显升高(P<0.05,P<0.01);与假手术组比较,正常组学习记忆能力、神经元细胞数量、PI3K/Akt/GSK-3β通路相关mRNA和蛋白表达水平差异均无统计学意义。结论:JuA可以改善VCI模型Objective:To investigate the effects of jujuboside A(JuA)on the learning and memory abilities and histopathological changes in the rat model of vascular cognitive impairment(VCI)and explore the potential mechanisms by which JuA treats VCI.Methods:A total of 50 male SPF-grade SD rats were randomized into a sham operation group(n=10),a blank control group(n=10),and a modeling group(n=30).The rats in the modeling group underwent bilateral carotid artery ligation(2-VO)for the modeling of VCI.After stabilization,the VCI rats were randomized into model,JuA(20 mg·kg^(-1)),and donepezil(0.45 mg·kg^(-1))groups.After 4 weeks of gavage,the novel object recognition and Morris water maze tests were conducted to evaluate the learning and memory abilities of rats.Nissl staining was employed to evaluate the morphology and number of hippocampal neurons.Real-time PCR was employed to measure the mRNA levels of glycogen synthase kinase-3β(GSK-3β),cAMP response element-binding protein(CREB),B cell lymphoma-2(Bcl-2),phosphatidylinositol 3-kinase(PI3K),and protein kinase B(Akt)in the hippocampal tissue.Western blot was employed to quantify the protein levels of GSK-3β,p-GSK-3β,p-CREB,Bcl-2,PI3K,p-PI3K,Akt,and p-Akt in the hippocampal tissue.Result:Compared with the sham operation group,the model group exhibited declines in the learning and memory abilities(P<0.01),neuronal damage and decreased neurons in the hippocampal CA1 region(P<0.01),up-regulation in the mRNA level of GSK-3β(P<0.01),and down-regulation in the mRNA levels of PI3K,Akt,CREB,and Bcl-2,as well as the protein levels of p-PI3K,p-Akt,p-GSK-3β,p-CREB,and Bcl-2(P<0.01).In comparison to the model group,both the JuA and donepezil groups demonstrated improvements in the learning and memory abilities(P<0.05,P<0.01),with reduced neuronal damage and increased neurons(P<0.05,P<0.01).In addition,the two groups showed down-regulation in the mRNA level of GSK-3β(P<0.01)and up-regulation in the mRNA levels of PI3K,Akt,CREB,and Bcl-2 and the protein levels of p-PI3K,p-Akt,p-GS

关 键 词：酸枣仁皂苷A 血管性认知障碍 学习记忆能力 细胞凋亡 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路 

分 类 号：R285[医药卫生—中药学] R282[医药卫生—中医学] R259