基于Rock2信号通路探讨黄芩苷对阿尔茨海默病小鼠学习记忆功能的影响  

Effects of Baicalin on Learning and Memory Function in Alzheimer′s Disease Mice Based on Rock2 Signaling Pathway

作　　者：黄燕[1] 骆飞飞[1] 唐卉[1] 周嫱[1] 徐朝义[1] HUANG Yan;LUO Feifei;TANG Hui;ZHOU Qiang;XU Chaoyi(The Third People′s Hospital of Chengdu,Chengdu 610000,Sichuan,China)

机构地区：[1]成都市第三人民医院,成都610000

出　　处：《中西医结合心脑血管病杂志》2022年第20期3709-3716,共8页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：四川省卫生健康委员会科研课题(No.19PJ169)。

摘　　要：目的构建阿尔茨海默病模型淀粉样前体蛋白(APP)及早老素-1(PS1)双转基因小鼠,观察黄芩苷对小鼠学习记忆功能改善的影响。方法体内实验将APP/PS1转基因小鼠分为模型组与黄芩苷组,同时将野生型小鼠作为对照组。黄芩苷组连续灌胃给药28 d,剂量60 mg/kg,其他两组灌胃等体积生理盐水。之后进行为期6 d的水迷宫实验。麻醉小鼠检测海马区兴奋性突触后电位。结束后处死小鼠,分离脑组织进行免疫组化染色,蛋白免疫印迹法(Western Blot)检测相关蛋白。体外实验应用PC12细胞,首先检测不同剂量β-淀粉样蛋白(Aβ)_(25-35)对PC12细胞存活率的影响,再检测黄芩苷对Aβ_(25-35)干预PC12细胞存活率的改善作用。将PC12细胞分为对照组、模型组、黄芩苷组与黄芩苷+溶血磷脂酸组。采用免疫荧光检测Rock2表达,Western Blot检测Caspase-3等凋亡相关蛋白表达。结果体内实验中,与模型组比较,经黄芩苷治疗的APP/PS1小鼠逃避潜伏期降低(P<0.05),穿越平台次数与靶象限停留时间增加(P<0.05)。黄芩苷可提高小鼠长时程电位(P<0.05),减少海马组织Aβ蛋白沉积(P<0.05)。Western Blot检测结果显示,黄芩苷减少了海马组织Rock2蛋白与Cleaved-Caspase-3蛋白表达(P<0.05),提高了Bcl-2/Bax比例(P<0.05)。体外实验中,200μmol/L黄芩苷显著提高了30μmol/L Aβ_(25-35)干预的PC12细胞存活率(P<0.05),抑制了Rho激酶(Rock2)蛋白与Cleaved-Caspase-3蛋白表达,提高Bcl-2/Bax比例(P<0.05)。溶血磷脂酸干预逆转了黄芩苷的作用,提高了Rock蛋白与Cleaved-Caspase-3蛋白表达,降低了Bcl-2/Bax比例。结论黄芩苷可治疗阿尔茨海默病,其机制是下调Rock2蛋白表达水平,减少Aβ沉积,减少细胞凋亡,提高长时程电位,改善小鼠学习与记忆功能障碍。Objective To observe the effect of baicalin on learning and memory function in Alzheimer′s disease model amyloid precursor protein(APP)gene and presenilin-1(PS1)double transgenic mice.Methods In vivo experiments,APP/PS1 transgenic mice were divided into model group and baicalin group,and wild-type mice were set as control group.The baicalin group was gavaged for 28 days with 60 mg/kg,and the other two groups were gavaged with an equal volume of normal saline.Afterwards,a 6-day water maze experiment was conducted.Excitatory postsynaptic potentials in mouse hippocampus were detected in anesthetized animals.In the end,the animals were sacrificed,and the brain tissues were isolated for immunohistochemical staining and Western Blot detection.PC12 cells were used for in vitro experiments.First,the effects of different doses ofβ-amyloid protein(Aβ)_(25-35) on the survival rate of PC12 cells were detected,and then the effect of baicalin on the survival rate of PC12 cells after Aβ_(25-35) intervention was detected.The PC12 cells were divided into control group,model group,baicalin group,and baicalin+lysophosphatidic acid group.The expression of Rock2 was detected by immunofluorescence,and the expression of apoptosis-related proteins such as Caspase-3 was detected by Western Blot.Results In in vivo experiment,compared with the model group,the escape latency of APP/PS1 mice treated with baicalin were decreased(P<0.05),and the number of crossing the platform and the residence time in the target quadrant were increased significantly(P<0.05).Baicalin could increase the long-term potential of mice(P<0.05)and decrease the deposition of Aβprotein in hippocampus(P<0.05).Western Blot detection showed that baicalin significantly reduced the expressions of Rock2 protein and Cleaved-Caspase-3 protein(P<0.05),and increased the expression ratio of Bcl-2/Bax(P<0.05)in hippocampus.In in vitro experiments,200μmol/L baicalin significantly increased the survival rate of PC12 cells intervened with 30μmol/L Aβ_(25-35) (P<0.05),inhibit

关 键 词：阿尔茨海默病 黄芩苷 RHO激酶 β-淀粉样蛋白 学习记忆功能 小鼠 实验研究 

分 类 号：R285.5[医药卫生—中药学]