人参总蛋白对Aβ25-35诱导SH-SY5Y细胞凋亡的机制研究  

Mechanism of Ginseng Total Protein on Apoptosis of SH-SY5Y Cells Induced by Aβ25-35

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作  者:郑丽娜 赵大庆 赵文学 乔巨慧 刘颖 刘美辰 ZHENG Li-na;ZHAO Da-qing;ZHAO Wen-xue;QIAO Ju-hui;LIU Ying;LIU Mei-chen(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,Jilin,China)

机构地区:[1]长春中医药大学吉林省人参科学研究院,吉林长春130117

出  处:《食品研究与开发》2019年第5期6-11,共6页Food Research and Developent

基  金:国家自然科学基金青年科学基金项目(81703663);国家重点研发计划(2017YFC1703100).

摘  要:对人参水溶性总蛋白对β-淀粉样蛋白所致人神经母细胞瘤SH-SY5Y损伤的影响及其作用机制进行研究。使用25μmol/L老化后Aβ25-35处理SH-SY5Y细胞建立阿尔兹海默病体外模型;用人参水溶性总蛋白(6.25、12.5、25μg/mL)预保护SH-SY5Y细胞12 h,随后加入25μmol/L的Aβ25-35共同孵育24 h;CCK-8法检测细胞活力;流式细胞术检测细胞凋亡、抑制活性氧(reactive oxygen species,ROS)水平和线粒体膜电位变化;紫外分光光度法检测三磷酸腺苷(adenosine 5'-triphosphate,ATP)、丙二醛(malondialdehyde,MDA)含量、超氧化物歧化酶(superoxide dismutase,SOD)活力;蛋白免疫印迹法检测细胞色素C释放以及凋亡相关蛋白B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)表达。结果显示SH-SY5Y细胞经Aβ25-35诱导后,细胞活力显著降低,凋亡程度严重增加(P<0.000 1);不同浓度的人参总蛋白可以挽救Aβ25-35诱导所致细胞活力降低和凋亡程度增加,并呈现浓度依赖性;人参总蛋白预处理可以降低Aβ25-35诱导的线粒体超氧化物升高,ROS产生,显著抑制Aβ25-35损伤导致的线粒体膜电位(mitochondrial membrane potential,MMP)降低和三磷酸腺苷水平的下降;人参总蛋白可抑制Aβ25-35所致的细胞色素C向胞质的释放,下调Bax表达、上调Bcl-2表达。The effects of ginseng water-soluble total protein on the damage of SH-SY5Y induced by amyloidβin human neuroblastoma were studied.In vitro model of Alzheimer's disease was established by treatment of SH-SY5Y cells with 25μmol/L aging Aβ25-35.SH-SY5Y cells were pre-protected with ginseng water-soluble total protein(6.25,12.5,25μg/mL)for 12 h.25μm/L of Aβ25-35 was added for 24 h.Cell viability was detected by CCK-8 method.Flow cytometry for apoptosis,reactive oxygen species(ROS)level and mitochondrial membrane potential were detected by flow cytometry.Ultraviolet spectrophotometric determination of adenosine 5'-triphosphate(ATP)and malondialdehyde(MDA),superoxide dismutase(SOD)activity.Western blotting was used to detect cytochrome C release and apoptosis-related protein B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)expression.The results showed that the cell viability was significantly decreased and the degree of apoptosis was significantly increased in SH-SY5Y cells induced by Aβ25-35(P<0.0001).Different concentrations of ginseng total protein could reduce the decrease of cell viability and the degree of apoptosis induced by Aβ25-35,and it was concentration dependent.Pretreatment with ginseng total protein could reduce the increase of mitochondrial superoxide induced by Aβ25-35,inhibit the production of ROS,and significantly inhibit the decrease of mitochondrial membrane potential(MMP)induced by Aβ25-35 injury,and decrease in the level of adenosine-triphosphate(ATP).Ginseng total protein inhibited the release of Cytochrome C to cytoplasm induced by Aβ25-35,down-regulated Bax expression and up-regulated Bcl-2 expression.

关 键 词:人参 阿尔兹海默病 细胞凋亡 β-淀粉样蛋白 线粒体 

分 类 号:Q421[生物学—神经生物学;生物学—生理学]

 

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