首页|基于AMPK/SIRT1通路探究苦参碱对阿尔茨海默病Aβ25-35诱导PC12细胞氧化损伤和凋亡的影响

基于AMPK/SIRT1通路探究苦参碱对阿尔茨海默病Aβ25-35诱导PC12细胞氧化损伤和凋亡的影响

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目的 探讨苦参碱对阿尔茨海默病β-淀粉样蛋白(Aβ)25-35诱导PC12细胞凋亡和氧化损伤的影响,并分析其与AMP活化蛋白激酶(AMPK)/沉默信息调节因子(SIRT1)通路的作用机制.方法 采用噻唑蓝(MTT)法检测不同浓度苦参碱对PC12细胞生长的影响,最终选择0.5、1.0和1.5 mmol/L苦参碱进行实验.将PC12细胞分为Con组(空白培养细胞),Aβ25-35组(20μmol/L的Aβ25-35处理细胞),Aβ25-35+Matrine-L组、Aβ25-35+Matrine-M组、Aβ25-35+Matrine-H组(20 μmol/L的Aβ25-35和0.5 mmol/L、1.0 mmol/L、2.0 mmol/L苦参碱处理细胞).用MTT法检测细胞增殖,流式细胞术检测细胞凋亡和活性氧(ROS)含量,酶联免疫吸附法检测超氧化的物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达水平,Western blotting检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved caspase-3)、AMPK/SIRT1通路相关蛋白表达.结果 与Con组相比,Aβ25-35组24 h、48 h的吸光度(OD)值、Bcl-2蛋白表达、SOD和GSH-Px活性均降低,凋亡率、Bax、cleaved caspase-3蛋白表达、ROS含量、IL-6、IL-1β、TNF-α表达增加,p-AMPK、SIRT1蛋白表达上调(均P<0.05);与Aβ25-35组相比,Aβ25-35+Matrine-L组、Aβ25-35+Matrine-M组、Aβ25-35+Matrine-H组24 h、48 h的OD值、Bcl-2蛋白表达、SOD和GSH-Px活性增加(P<0.05),凋亡率、Bax、cleaved caspase-3蛋白表达、ROS含量、IL-6、IL-1β、TNF-α表达降低,p-AMPK、SIRT1蛋白表达下调(P<0.05).结论 苦参碱可能通过抑制AMPK/SIRT1通路激活,减轻Aβ25-35诱导的PC12细胞凋亡和氧化损伤.
Effect of matrine on oxidative damage and apoptosis of PC12 cells via AMPE/SIRT1 pathway in Alzheimer disease induced by Aβ25-35
Objective To investigate the effect of matrine on apoptosis and oxidative damage of PC12 cells induced byβ-amyloid protein(Aβ)25-35 in Alzheimer disease and to analyze the mechanism of its interaction with the AMP-activated protein kinase(AMPK)/silenced information regulator(SIRT1)pathway.Methods The effect of matrine on the growth of PC12 cells was determined using MTT.Matrine concentrations of 0.5,1.0,and 1.5 mmol/L were selected for the experiment.PC12 cells were divided into the Con group(blank culture cells),the Aβ25-35 group(20 μmol/L Aβ25-35 treated cells),the Aβ25-35+Matrine-L group,the Aβ25-35+Matrine-M group,and the Aβ25-35+Matrine-H group(20μmol/L Aβ25-35 were treated with 0.5 mmol/L,1.0 mmol/L,and 2.0 mmol/L matrine).Cell proliferation was detected using MTT.The detection of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)using enzyme-linked immunosorbent assay,expression of B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),cleaved caspase-3 containing cysteine,and AMPK/SIRT1 pathway-associated proteins were detected using Western blotting.Results Compared to the Con group,OD value,Bcl-2 protein expression,SOD,and GSH-Px activity decreased at 24 h and 48 h;the apoptosis rate,Bax,cleaved caspase-3 protein expression,ROS content,IL-6,IL-1β,and TNF-α expression increased,and P-AMPK and SIRT1 protein expression were up-regulated in the Aβ25-35 group(all P<0.05).Compared to the Aβ25-35 group,OD value,Bcl-2 protein expression,SOD,and GSH-Px activities in the Aβ25-35+Matrine-L group,the Aβ25-35+Matrine-M group,and the Aβ25-35+Matrine-H group increased at 24 h and 48 h.Apoptosis rate,Bax,cleaved caspase-3 protein expression,and ROS content decreased.IL-6,IL-1β,and TNF-αexpression decreased,and p-AMPK,and SIRT1 protein expression were down-regulated(all P<0.05).Conclusion Matrine may reduce the apoptosis and oxidative damage of PC12 cells induced by Aβ25-35 by inhibiting the AMPK/SIRT1 pathway.

matrineAlzheimer diseasecell apoptosisoxidative stressinflammatory response

刘景祎、李富慧、宋彦

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南阳市中心医院神经内科,河南 南阳 473000

南阳市第二人民医院神经内科,河南 南阳 473000

苦参碱 阿尔茨海默病 细胞凋亡 氧化应激 炎症反应

河南省医学科技攻关计划(联合共建)项目

LHGJ20191467

2024

中国医科大学学报
中国医科大学

中国医科大学学报

CSTPCD北大核心
影响因子:1.421
ISSN:0258-4646
年,卷(期):2024.53(8)