摘要
目的:探讨脱蛋白后琐琐葡萄多糖(DVTP)对Aβ1-42 诱导的阿尔兹海默症细胞模型氧化损伤的影响.方法:将HT22细胞分为对照组,模型组(Aβ1-42)及Aβ1-42+DVTP(20、40、80 μg/mL)组,使用CCK8检测细胞存活率,在光镜、透射电镜下观察细胞形态和线粒体结构变化,DCFH-DA和JC-10荧光探针法测定细胞中活性氧(ROS)及线粒体膜电位水平,同时利用RT-qPCR和Western blot分析Keap1、Nrf2、HO-1、NQO1、GCLC的mRNA和蛋白表达水平.结果:与对照组相比,模型组细胞存活率下降,细胞形态及线粒体结构损伤程度较高;ROS水平升高,线粒体膜电位下降;Keap1 mRNA及蛋白表达水平升高,Nrf2、HO-1、NQO1、GCLC表达水平降低.同模型组相比,DVTP预处理后细胞存活率显著提升,细胞形态及线粒体结构损伤程度较低;ROS水平降低,线粒体膜电位升高;Keapl mRNA及蛋白表达水平下降,Nrf2、HO-1、NQO1、GCLC表达水平升高.结论:DVTP可通过调节抗氧化酶的表达水平改善Aβ1-42 诱导的氧化损伤,有望成为一种治疗AD的候选药物.
Abstract
Objective:To investigate the effect of deproteinized polysaccharides from Vitis vinifera L.Suosuo(DVTP)on ox-idative damage induced by Aβ1-42 in Alzheimer's disease cell model.Methods:HT22 cells were divided into the control group,the model group(Aβ1-42),and the Aβ1-42+DVTP(20,40,80 μg/mL)groups.Cell viability was measured using the CCK8 assay.Cell morphology and mitochondrial structure changes were observed under light and transmission electron microscopy.ROS levels and mitochondrial membrane potential were assessed using DCFH-DA and JC-10 fluorescent probes,respectively.RT-qPCR and Western blot were used to analyze the mRNA and protein expression levels of Keap1,Nrf2,HO-1,NQO1,and GCLC.Results:Compared to the control group,the model group exhibited decreased cell viability and increased damage to cell morphology and mi-tochondrial structure.ROS levels were elevated,and mitochondrial membrane potential was reduced.The mRNA and protein ex-pression levels of Keap1 were increased,while those of Nrf2,HO-1,NQO1,and GCLC were decreased.In comparison to the model group,DVTP pretreatment significantly improved cell viability,reduced damage to cell morphology and mitochondrial structure,decreased ROS levels,and increased mitochondrial membrane potential.The mRNA and protein expression levels of Keap1 were reduced,while those of Nrf2,HO-1,NQO1,and GCLC were increased.Conclusion:DVTP can mitigate Aβ1-42-in-duced oxidative damage by regulating the expression levels of antioxidant enzymes,suggesting its potential as a candidate drug for the treatment of Alzheimer's disease.
基金项目
国家自然科学基金(81960764)
新疆医科大学博士科研启动金项目(XJMUYJ20210603)
万方数据