海松子提取物减轻阿尔茨海默病线虫Aβ毒性及机制探讨
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摘要
目的:本实验探讨海松子提取物(EKps)对阿尔茨海默病(AD)转基因秀丽隐杆线虫体内β-淀粉样蛋白(β-amyloid,Aβ)毒性的影响及作用机制。方法:线虫设空白对照组、EKps组和EGb761阳性对照组。利用瘫痪实验比较各组AD线虫的Aβ毒性,硫磺素s染色测定各组AD线虫体内Aβ表达量的变化,胡桃醌诱导的氧化应激实验分析线虫的抗氧化应激能力,用标记谷胱甘肽s转移酶-4(GST-4::GFP)的线虫观察线虫体内GST-4表达量。结果:较之对照组,EKps延缓了由Aβ毒性导致的AD线虫瘫痪(P <0. 000 1),减少了AD线虫体内Aβ的沉积(P <0. 000 1),也增强了胡桃醌诱导下线虫的抗氧化应激能力(P=0. 001),并增加了线虫体内GST-4的表达(P <0. 000 1)。结论:EKps具有减少Aβ聚集从而减轻AD线虫体内Aβ毒性的作用,机制可能与增强线虫的抗氧化应激能力相关。
Objective: This study aimed to discuss the effect of the extract of korean pine seed (EKps) on the toxicity of β-amyloid (Aβ) in an Alzheimer's disease (AD) model of Caenorhabditis elegans (C. elegans) and explore its mechanism. Methods: A blank control group,EKps group and EGb761 positive control group were set.The Aβ toxicity in the AD C. elegans was compared by paralysis test. Changes of Aβ expression in the AD C. elegans in each group were measured by thioflavin-s staining. The oxidative stress experiment was induced by juglone to observe whether EKps can enhance the ability of anti-oxidative stress. C. elegans labeled with glutahtione s transferase-4 (GST-4: : GFP) were used for determining the expression of GST-4. Results: Aβ-induced paralysis in the AD C. elegans is alleviated by EKps (P < 0. 000 1). EKps also reduced the desposits of Aβ in the AD C. elegans (P < 0. 000 1) and enhanced the ability of anti-oxidative stress (P = 0. 001). In addition,EKps increased the expression of GST-4 in C. elegans (P < 0. 000 1). Conclusion: EKps alleviated the toxicity of Aβ in the AD C. elegans through reducing the deposition of Aβ,which may be related to enhancing the ability of C. elegans to resist oxidative stress.
Objective: This study aimed to discuss the effect of the extract of korean pine seed (EKps) on the toxicity of β-amyloid (Aβ) in an Alzheimer's disease (AD) model of Caenorhabditis elegans (C. elegans) and explore its mechanism. Methods: A blank control group,EKps group and EGb761 positive control group were set.The Aβ toxicity in the AD C. elegans was compared by paralysis test. Changes of Aβ expression in the AD C. elegans in each group were measured by thioflavin-s staining. The oxidative stress experiment was induced by juglone to observe whether EKps can enhance the ability of anti-oxidative stress. C. elegans labeled with glutahtione s transferase-4 (GST-4: : GFP) were used for determining the expression of GST-4. Results: Aβ-induced paralysis in the AD C. elegans is alleviated by EKps (P < 0. 000 1). EKps also reduced the desposits of Aβ in the AD C. elegans (P < 0. 000 1) and enhanced the ability of anti-oxidative stress (P = 0. 001). In addition,EKps increased the expression of GST-4 in C. elegans (P < 0. 000 1). Conclusion: EKps alleviated the toxicity of Aβ in the AD C. elegans through reducing the deposition of Aβ,which may be related to enhancing the ability of C. elegans to resist oxidative stress.
引文
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[17]董雯,王守安,陈秀娇,等.白藜芦醇在阿尔茨海默病防治中作用机制研究进展[J].中国现代应用药学,2018,35(11):1745-1748.
[18]杨婷婷,周红静,曾晨叶,等.新型天麻素衍生物对阿尔茨海默病模型小鼠的保护作用[J].中国现代应用药学,2019,36(5):537-541.
[19]李林,臧彩霞,郑远鹏,等.中药栀子提取物GJ-4对亚急性衰老致痴呆小鼠学习记忆的保护作用[J].中国新药杂志,2017,26(11):1289-1295.
[20]WU Y,WU Z,BUTKO P,et al.Amyloid-beta-induced pathological behaviors are suppressed by Ginkgo biloba extract EGb761 and ginkgolides in transgenic Caenorhabditis elegans[J].JNeurosci,2006,26(50):13102-13113.
[2]XU J,WANG J,WIMO A,et al.The economic burden of dementia in China,1990-2030:implications for health policy[J].B World Health Organ,2017,95(1):18-26.
[3]APETZ N,MUNCH G,GOVINDARAGHAVAN S,et al.Natural compounds and plant extracts as therapeutics against chronic inflammation in Alzheimer's disease---a translational perspective[J].CNS Neurol Disorder,2014,13(7):1175-1191.
[4]缪希雍著.郑金生校注.神农本草经疏[M].北京:中医古籍出版社,2002:685.
[5]国家中医药管理局《中华本草》编委会.中华本草[M].上海:上海科学技术出版社,1999:2298.
[6]TAKAHASHI A,WATANABE T,FUJITA T,et al.Green tea aroma fraction reduces beta-amyloid peptide-induced toxicity in Caenorhabditis elegans transfected with human beta-amyloid minigene[J].Biosci Biotech Bioch,2014,78(7):1206-1211.
[7]ZHANG XG,WANG X,ZHOU TT,et al.Scorpion venom heatresistant peptide protects transgenic Caenorhabditis elegans from beta-amyloid toxicity[J].Front Pharmacol,2016,7:227.
[8]LIU H,LIANG F,SU W,et al.Lifespan extension by n-butanol extract from seed of Platycladus orientalis in Caenorhabditis elegans[J].J Ethnopharmacol,2013,147(2):366-372.
[9]O'REILLY LP,LUKE CJ,PERLMUTTER DH,et al.C.elegans in high-throughput drug discovery[J].Adv Drug Deliv Rev,2014(69-70):247-253.
[10]CHEN Z,ZHONG C.Oxidative stress in Alzheimer's disease[J].Neurosci Bull,2014,30(2):271-281.
[11]WANG X,WANG W,LI L,et al.Oxidative stress and mitochondrial dysfunction in Alzheimer's disease[J].Biochim Biophys Acta,2014,1842(8):1240-1247.
[12]HATANAKA H,HANYU H,HIROSE D,et al.Peripheral oxidative stress markers in individuals with Alzheimer's disease with or without cerebrovascular disease[J].J Am Geriatr Soc,2015,63(7):1472-1474.
[13]DUBININA EE,SCHEDRINA LV,NEZNANOV NG,et al.Oxidative stress and its effect on cells functional activity of alzheimer's disease[J].Biomed Khim,2015,61(1):57-69.
[14]LUQUE-CONTRERAS D,CARVAJAL K,TORAL-RIOS D.Oxidative stress and metabolic syndrome:cause or consequence of Alzheimer's disease?[J].Oxid Med Cell Longev,2014,2014:497802.
[15]MAO S,ZHOU F,HUANG W,et al.The effect of traditional stir-frying process on hydrophilic and lipophilic antioxidant capacities of pine nut kernels[J].Int J Food Sci Nutr,2015,66(8):873-880.
[16]ZHANG J,LIN W,WU R,et al.Mechanisms of the active components from Korean pine nut preventing and treating d-galactoseinduced aging rats[J].Biomed Pharmacother,2018(103):680-690.
[17]董雯,王守安,陈秀娇,等.白藜芦醇在阿尔茨海默病防治中作用机制研究进展[J].中国现代应用药学,2018,35(11):1745-1748.
[18]杨婷婷,周红静,曾晨叶,等.新型天麻素衍生物对阿尔茨海默病模型小鼠的保护作用[J].中国现代应用药学,2019,36(5):537-541.
[19]李林,臧彩霞,郑远鹏,等.中药栀子提取物GJ-4对亚急性衰老致痴呆小鼠学习记忆的保护作用[J].中国新药杂志,2017,26(11):1289-1295.
[20]WU Y,WU Z,BUTKO P,et al.Amyloid-beta-induced pathological behaviors are suppressed by Ginkgo biloba extract EGb761 and ginkgolides in transgenic Caenorhabditis elegans[J].JNeurosci,2006,26(50):13102-13113.