基于磷脂酰肌醇3-激酶/蛋白激酶B信号通路探讨黄芪甲苷对过氧化氢诱导的PC12细胞凋亡的保护作用  

Effects of astragaloside Ⅳ against hydrogen peroxide-induced apoptosis of PC12 cells via the PI3K/Akt signaling pathway

作　　者：邓刚 周康 付建辉 DENG Gang;ZHOU Kang;FU Jianhui(Department of Neurosurgery,Tonglu County First People's Hospital,Hangzhou,Zhejiang 311500,China)

机构地区：[1]杭州市桐庐县第一人民医院神经外科,杭州311500

出　　处：《浙江中西医结合杂志》2024年第7期604-610,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的研究黄芪甲苷对过氧化氢(H_(2)O_(2))诱导的大鼠肾上腺嗜铬细胞瘤细胞(PC12)凋亡的保护作用及机制。方法本实验分为正常组、模型组及黄芪甲苷低、中、高(0.4、0.6、0.8μmol/L)剂量组,使用不同浓度黄芪甲苷干预经H_(2)O_(2)诱导的PC12细胞。使用2,7-二氯荧光素二乙酸酯(DCFH-DA)荧光探针染色检测PC12细胞内活性氧(ROS)含量,赫斯特荧光染色液(Hoechst33258)染色检测PC12细胞相对凋亡率,蛋白质印迹法(Western blot)检测PC12细胞凋亡相关蛋白及磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路关键蛋白的表达。结果与正常组比较,模型组PC12细胞经H_(2)O_(2)诱导后细胞存活率[(48.53±8.31)%比(100.00±5.61)%,P<0.01]显著下降,细胞内ROS相对含量[(579.77±22.31)%比(100.00±9.76)%,P<0.01]、细胞相对凋亡率[(453.80±49.10)%比(100.00±10.89)%,P<0.01]显著升高,凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)相对表达率[(31.87±3.46)%比(100.00±12.38)%,P<0.01]显著下降,半胱氨酸蛋白酶-3(Caspase-3)[(205.73±9.40)%比(100.00±10.33)%,P<0.01]、Bcl-2相关x蛋白(Bax)[(302.80±19.50)%比(100.00±10.61)%,P<0.01]相对表达率显著升高,磷酸化磷脂酰肌醇3-激酶(p-PI3K)[(46.07±3.78)%比(100.00±11.46)%,P<0.01]、磷酸化蛋白激酶B(p-Akt)[(35.70±2.26)%比(100.00±9.66)%,P<0.01]相对表达率显著下降。与模型组比较,经黄芪甲苷干预后的黄芪甲苷中、高剂量组细胞存活率[(65.53±5.30)%、(73.13±6.16)%比(48.53±8.31)%,P<0.05或P<0.01]显著升高,细胞内ROS相对含量[(473.23±40.24)%、(342.27±36.81)%比(579.77±22.31)%,P<0.05或P<0.01]显著下降,细胞相对凋亡率[(366.77±47.65)%、(262.33±48.01)%比(453.80±49.10)%,P<0.05或P<0.01]显著下降,Bcl-2相对表达率[(44.43±6.13)%、(68.57±9.93)%比(31.87±3.46)%,P<0.05或P<0.01]显著升高,Caspase-3[(152.57±7.74)%、(114.40±6.27)%比(205.73±9.40)%,P<0.05或P<0.01]及Bax[(247.67±18.50)%、(182.73±24.14)%比(302.80±19.50)%,P<0.05或P<0.01]相对�Objective To explore the protective effects and mechanisms of astragaloside Ⅳ on hydrogen peroxide(H_(2)O_(2))-induced apoptosis in rat adrenal pheochromocytoma cells(PC12).Methods The experiment was divided into normal,model,and low,medium,and high-dose astragalosideⅣ(0.4,0.6,0.8μmol/L)groups.Different concentrations of astragalosideⅣwere used to treat H_(2)O_(2)-induced PC12 cells.Reactive oxygen species(ROS)levels in PC12 cells were detected using 2,7-dichlorofluorescein diacetate fluorescent probe staining.The apoptosis rate of PC12 cells was assessed using Hoechst 33258 fluorescent staining.Western blot analysis was used to detect the expression of apoptosis-related proteins and key proteins in phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway in PC12 cells.Results Compared with the normal group,after being induced by H_(2)O_(2),cell survival rates of the PC12 cells in the model group were significantly decreased[(48.53±8.31)%vs.(100.00±5.61)%,P<0.01].The levels of intracellular ROS[(579.77±22.31)%vs.(100.00±9.76)%,P<0.01]and the relative apoptosis rate[(453.80±49.10)%vs.(100.00±10.89)%,P<0.01]were significantly increased.The expression of apoptosis-related protein Bcl-2 was significantly decreased[(31.87±3.46)%vs.(100.00±12.38)%,P<0.01],while the expression of Caspase-3[(205.73±9.40)%vs.(100.00±10.33)%,P<0.01]and Bax[(302.80±19.50)%vs.(100.00±10.61)%,P<0.01]were significantly increased.Additionally,the expression levels of p-PI3K[(46.07±3.78)%vs.(100.00±11.46)%,P<0.01]and p-Akt[(35.70±2.26)%vs.(100.00±9.66)%,P<0.01]were significantly decreased.Compared with the model group,after the intervention of astragalosideⅣ,the cell survival rate of the medium and high dose astragaloside Ⅳ groups was significantly increased[(65.53±5.30)%,(73.13±6.16)%vs.(48.53±8.31)%,P<0.05 or P<0.01].The levels of intracellular ROS were significantly decreased[(473.23±40.24)%,(342.27±36.81)%vs.(579.77±22.31)%,P<0.05 or P<0.01],and the relative cell apoptosis rates were signific

关 键 词：脑卒中 细胞凋亡 磷脂酰肌醇3-激酶/蛋白激酶B 黄芪甲苷 PC12 

分 类 号：R285[医药卫生—中药学]