朱江华,范焕琼
Zhu Jiang Hua,Fan Huan Qiong

• 1:广东省佛山市妇幼保健院药学部
• 2:广东省佛山市南海区第五人民医院药学科

摘要(Abstract)：

目的探讨白藜芦醇拮抗Aβ_(1-42)诱导的体外培养神经干细胞C17.2凋亡的可能机制。方法实验分组分别为:空白对照组、Aβ_(1-42)(10μmol·L~(-1))处理组、白藜芦醇干预组(白藜芦醇1,5,10,20μmol·L~(-1)+Aβ_(1-42)10μmol·L~(-1))。采用MTT法检测细胞活力;流式细胞仪检测细胞凋亡率;采用Western blotting法分别检测热休克蛋白75(Hsp75)和活化型caspase-3蛋白的表达水平。结果与Aβ_(1-42)处理组相比,白藜芦醇预处理组呈剂量依赖性地显著提高神经干细胞的存活率(P<0.05)和凋亡率(P<0.05);同样地,显著上调C17.2细胞Hsp75蛋白的表达和降低其活化型caspase-3蛋白水平(P<0.05)。而单独使用白藜芦醇对正常C17.2细胞活性影响的差异无统计学意义(P>0.05)。结论白藜芦醇对Aβ_(1-42)诱导的C17.2细胞损伤具有明显保护作用,其作用机制可能与上调C17.2细胞Hsp75蛋白的表达水平和抑制caspase-3蛋白介导的凋亡途径从而降低细胞凋亡有关。
OBJECTIVE To investigate the possible mechanism of resveratrol weakening the apoptosis of neural stem cells C17.2induced by Aβ_(1-42) in vitro. METHODS In this paper,we set six experimental groups,such as control group,Aβ_(1-42)( 10 μmol·L~(-1))treating group,resveratrol pretreating group( 1,5,10,20 μmol·L~(-1)+ Aβ_(1-42)10 μmol·L~(-1)). Cell viability was measured by MTT assay;apoptotic rate was detected by Flow cytometry,western blotting was used to test the expression of Hsp75 and activating caspase-3 protein.RESULTS Compared with Aβ_(1-42) treatment group,resveratrol pretreament group significantly increased the C17. 2 cells survival rate( P>0. 05) and apoptosis rate( P>0. 05) in a dose-dependent manner. Similarly,resveratrol significantly ameliorate the expression level of Hsp75 protein and reduced the level of activated caspase-3 protein in C17.2 cells in Aβ_(1-42) treatment group. However,there was not significant effect of resveratrol on the viability of C17. 2 cells( P > 0. 05). CONCLUSION Resveratrol has an obviously protective effect on Aβ_(1-42) induced damage of C17. 2 cells. The possible mechanism was that resveratrol up-regulated the expression of Hsp75 protein in C17.2 cells and inhibited the caspase-3 protein-associated apoptotic pathway.

关键词(KeyWords)： 白藜芦醇;阿尔茨海默病;热休克蛋白75;神经干细胞
resveratrol;Alzheimer's disease;heat shock protein-75;nueral stem cell

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 朱江华,范焕琼
Zhu Jiang Hua,Fan Huan Qiong

参考文献(References)：

• [1]Selkoe,D J.Developing Preventive Therapies for Chronic Diseases:Lessons Learned from Alzheimer's Disease[J].Nutrition Reviews,2008,65(12):239-243.
• [2]Sadigh-Eteghad S,Sabermarouf B,Majdi A,et al.,Amyloid-beta:a crucial factor in Alzheimer's disease[J].Medical Principles&Practice International Journal of the Kuwait University Health Science Centre,2015,24(1):1.
• [3]葛宇松,尹琳,滕伟禹,等.β-淀粉样蛋白诱导大鼠海马神经元凋亡及海马亲环素A表达的变化[J].中华神经医学杂志,2012,11(4):337-341.
• [4]杨英,张倩,王智斌,等.线粒体乙醛脱氢酶2对β淀粉样蛋白所致神经元损伤的作用及机制[J].中国老年学,2015,(5):1309-1311.
• [5]Du H,Guo L,Fang F,et al.Cyclophilin D deficiency attenuates mitochondrial and neuronal perturbation and ameliorates learning and memory in Alzheimer's disease[J].Nature Medicine,2008,14(10):1097-1105.
• [6]Ruggiero A,Aloni E,Korkotian E,et al.Loss of forebrain MTCH2decreases mitochondria motility and calcium handling and impairs hippocampal-dependent cognitive functions[J].Scientific Reports,2017,7:44401.
• [7]Rege S D,Geetha T,Griffin G D,et al.Neuroprotective effects of resveratrol in Alzheimer disease pathology[J].Frontiers in Aging Neuroscience,2014,6(6):218.
• [8]Costantino E,Maddalena F,Calise S,et al.TRAP1,a novel mitochondrial chaperone responsible for multi-drug resistance and protection from apoptotis in human colorectal carcinoma cells[J].Cancer Letters,2009,279(1):39-46.
• [9]Orallo F.Comparative studies of the antioxidant effects of cis-and trans-resveratrol[J].Current Medicinal Chemistry,2006,13(1):87-98.
• [10]Das S,Falchi M,Bertelli A,et al.Attenuation of ischemia/reperfusion injury in rats by the anti-inflammatory action of resveratrol[J].Arzneimittel-Forschung,2006,56(10):700.
• [11]杨迎暴,罗景慧,徐江平,等.白黎芦醇对f MLP诱导中性白细胞与人脐静脉内皮细胞粘附的抑制作用及机制[J].中国药理学通报,2007,23(12):1588-1593.
• [12]Baur J A,Sinclair D A.Therapeutic potential of resveratrol:the in vivo evidence[J].Nature Reviews Drug Discovery,2006,5(6):493.
• [13]Voloboueva L A,Lee S W,Emery J F,et al.Mitochondrial protection attenuates inflammation-induced impairment of neurogenesis in vitro and in vivo[J].Journal of Neuroscience the Official Journal of the Society for Neuroscience,2010,30(37):12242.
• [14]Bastianetto S C,Ménard,Quirion R.Neuroprotective action of resveratrol[J].Biochimica Et Biophysica Acta,2015,1852(6):1195.
• [15]Mendiola-Precoma J,Berumen L C,Padilla K,et al.Therapies for Prevention and Treatment of Alzheimer's Disease[J].Biomed Res Int,2016,2016(3):1-17.
• [16]Lundqvist J,El A J,Svensson C,et al.Optimisation of culture conditions for differentiation of C17.2 neural stem cells to be used for in vitro toxicity tests[J].Toxicology in vitro,2013,27(5):1565.
• [17]Chen S,Luo M,Zhao Y,et al.Fasudil Stimulates Neurite Outgrowth and Promotes Differentiation in C17.2 Neural Stem Cells by Modulating Notch Signalling but not Autophagy[J].Cellular Physiology&Biochemistry International Journal of Experimental Cellular Physiology Biochemistry&Pharmacology,2015,36(2):531-541.
• [18]段答,姜淼,叶玉勤,等.嗅鞘细胞分泌蛋白SC1促进神经元突触形成[J].中华神经外科疾病研究杂志,2015,14(3):204-207.
• [19]Xiang F.,Huang Y S,Shi X H,et al.Mitochondrial chaperone tumour necrosis factor receptor-associated protein 1 protects cardiomyocytes from hypoxic injury by regulating mitochondrial permeability transition pore opening[J].Febs Journal,2010,277(8):1929-1938.
• [20]Voloboueva L A,Duan M,Ouyang Y B,et al.Overexpression of mitochondrial Hsp70/Hsp75 protects astrocytes against ischemic injury in vitro[J].Journal of Cerebral Blood Flow&Metabolism Official Journal of the International Society of Cerebral Blood Flow&Metabolism,2008,28(5):1009.
• [21]Wang Y,Lin J,Chen Q,et al.Overexpression of mitochondrial Hsp75 protects neural stem cells against microglia-derived soluble factor-induced neurotoxicity by regulating mitochondrial permeability transition pore openingin vitro[J].International Journal of Molecular Medicine,2015,36(6):1487-1496.