Study on the Anti-aging Effect of Cherry Ethanol Extract Based on Caenorhabditis elegans Model
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摘要: 本文以胶东地区常见的两种樱桃品种—红灯和黄蜜为研究对象，探究并比较了樱桃的抗衰老作用，为樱桃的进一步开发利用提供依据。对两种樱桃的果肉和果核分别进行醇溶提取并初步明确成分，以DPPH自由基和羟自由基清除能力为指标评价四种成分的抗氧化活性并探究其抑制β淀粉样蛋白(Aβ）聚集的能力。以秀丽隐杆线虫为模型，测定四种成分对其寿命、运动能力、生殖能力、瘫痪缓解能力的作用效果和抑制Aβ沉积情况。结果表明：两种樱桃果肉的抗氧化活性突出，样品浓度在1 mg/mL时，黄蜜和红灯的DPPH自由基清除率分别为80.88%和83.19%，羟自由基清除率分别为88.19%和92.85%。果核相对较低，黄蜜和红灯的DPPH自由基清除率分别为20.76%和44.93%，羟自由基清除率分别为28.65%和38.75%。四种样品均对Aβ表现出抑制作用，且在线虫实验中，与空白组相比，四种样品组的线虫寿命能够延长2~4 d，增强其运动能力，对生殖能力无损害且降低线虫瘫痪率，减少Aβ沉积。综上，四种样品能够通过清除自由基，抑制Aβ的产生从而产生一定的抗衰老效果，具有潜在的应用价值。Abstract: In this paper, the anti-aging effects of two varieties of cherry, Red Light and Yellow Honey in Jiaodong region were investigated and compared , for further development and utilization of cherries. The components of pulp and kernels in the two kinds of cherries were extracted by ethanol and the antioxidant activity was evaluated by the scavenging ability of DPPH and hydroxyl radicals, the ability to inhibit the aggregation of β-amyloid protein (Aβ) was investigated. The effects of four components on the life span, locomotor ability, reproductive ability, paralysis relief ability and the inhibition of Aβ deposition of Caenorhabditis elegans were studied. The results showed that antioxidant activity of two cherry pulp was outstanding, with DPPH radical scavenging rates of 80.88% and 83.19% and hydroxyl radical scavenging rates of 88.19% and 92.85% for Yellow Honey and Red Light, respectively, when the sample concentration was 1 mg/mL. While the antioxidant activity of fruit kernels was relatively low, with DPPH and hydroxyl radical scavenging rates of Yellow Honey and Red Light were 20.76% and 44.93%, 28.65% and 38.75%, respectively. Compared with the blank group, the life span of Caenorhabditis elegans in the four groups was prolonged by 2~4 days and their locomotor ability was enhanced, and it did not damage the reproduction ability and reduced the paralysis rate of Caenorhabditis elegans and Aβ deposition. In conclusion, the four samples were able to produce certain anti-aging effects by scavenging free radicals and inhibiting the production of Aβ, which have potential applications.
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Key words:
- cherry /
- antioxidant /
- beta-amyloid /
- anti-aging /
- Caenorhabditis elegans
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图 1 四种样品醇提物中总多酚和总黄酮含量
Figure 1. Total polyphenols and total flavonoids in four sample ethanol extracts
注：图中不同小写字母表示差异显著 P<0.05；图7同。
图 2 四种样品醇提物清除DPPH能力和羟自由基（·OH）能力
Figure 2. The ability of four sample ethanol extracts extracts to scavenge DPPH and hydroxyl radicals (·OH)
图 3 ThT荧光检测四种样品醇提物对Aβ的聚集影响
Figure 3. ThT fluorescence detection of the effect of four sample alcohol extracts on the aggregation of Aβ
注：具有不同侧标的数据之间具有显著性差异，P<0.05。
图 4 四种样品醇提物对秀丽隐杆线虫寿命的影响
Figure 4. Effects of four sample alcohol extracts on the lifespan of Caenorhabditis elegans
图 5 四种样品醇提物对秀丽隐杆线虫运动能力的影响
Figure 5. Effects of four sample alcohol extracts on motility of Caenorhabditis elegans
图 6 四种样品醇提物对秀丽隐杆线虫生殖能力的影响
Figure 6. Effects of four sample alcohol extracts on reproductive capacity of Caenorhabditis elegans
注：*表示与同一天空白组相比具有显著性差异，P<0.05。
图 7 四种样品醇提物对秀丽隐杆线虫瘫痪的影响
Figure 7. Effects of four sample alcohol extracts on paralysis of Caenorhabditis elegans
图 8 四种样品醇提物对秀丽隐杆线虫体内Aβ蛋白沉积情况的影响（20 µm）
Figure 8. Effect of alcoholic extracts of four samples on the deposition of Aβ protein in Caenorhabditis elegans (20 µm)
注：A：Aβ蛋白在空白组线虫体内沉积情况；B：Aβ蛋白在样品组线虫体内沉积情况。
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[1] 张明. 温室甜樱桃品种优选和高效栽培技术研究[D]. 泰安: 山东农业大学, 2014ZHANG M. Research on variety selection and high-efficiency cultivation technology of greenhouse sweet cherry[D]. Tai'an: Shandong Agricultural University, 2014. [2] 高子飞. 樱桃中几种多酚类物质清除自由基机理研究[D]. 烟台: 烟台大学, 2019.GAO Z F. Study on the free radical scavenging mechanism of several polyphenols in cherries[D]. Yantai: Yantai University, 2019. [3] 刘英, 雒新艳, 谭延庆, 等. 寒亭区主要甜樱桃品种果实品质比较研究[J]. 北方果树,2021(2):10−13. [LIU Y, LUO X Y, TAN Y, et al. Comparative study on fruit quality of main sweet cherry varieties in Hanting District[J]. North Fruit Trees,2021(2):10−13. doi: 10.16376/j.cnki.bfgs.2021.02.003[LIU Y, LUO X Y, TAN Y, et al. Comparative study on fruit quality of main sweet cherry varieties in Hanting District[J]. North Fruit Trees, 2021(2): 10-13. doi: 10.16376/j.cnki.bfgs.2021.02.003. [4] 梁秋萍, 严学迎. 基于熵权TOPSIS法的不同品种甜樱桃营养品质综合评价[J]. 食品研究与开发,2021,42(16):59−64. [LIANG Q P, YAN X Y. Comprehensive evaluation of nutritional quality of different varieties of sweet cherry based on entropy weight TOPSIS method[J]. Food Research and Development,2021,42(16):59−64. doi: 10.12161/j.issn.1005-6521.2021.16.009[LIANG Q P, YAN X Y. Comprehensive evaluation of nutritional quality of different varieties of sweet cherry based on entropy weight TOPSIS method[J]. Food Research and Development, 2021, 42(16): 59-64. doi: 10.12161/j.issn.1005-6521.2021.16.009 [5] 闫慧明. 樱桃多酚提取物对DSS诱导的小鼠溃疡性结肠炎保护作用研究[D]. 重庆: 西南大学, 2021YAN H M. The protective effect of cherry polyphenol extract on DSS-induced ulcerative colitis in mice[D]. Chongqing: Southwest University, 2021. [6] BECONCINI D, FELICE F, FABIANO A, et al. Antioxidant and anti-inflammatory properties of cherry extract: Nanosystems-based strategies to improve endothelial function and intestinal absorption[J]. Foods,2020,9(2):207. doi: 10.3390/FOODS9020207 [7] 董渭雪. 樱桃黄酮组分及降尿酸作用研究[D]. 汉中: 陕西理工大学, 2020DONG W X. Study on cherry flavonoid components and urate-lowering effects[D]. Hanzhong: Shaanxi University of Technology, 2020. [8] 郭遥遥, 刘洋, 王沙沙, 等. 樱桃核提取物抗疲劳、耐缺氧及镇痛作用的研究[J]. 青岛科技大学学报(自然科学版),2018,39(4):28−32. [GUO Y Y, LIU Y, WANG S S, et al. Study on anti-fatigue, hypoxia tolerance and analgesic effects of cherry kernel extract[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition),2018,39(4):28−32. doi: 10.16351/j.1672-6987.2018.04.004[GUO Y Y, LIU Y, WANG S S, et al. Study on anti-fatigue, hypoxia tolerance and analgesic effects of cherry kernel extract[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition), 2018, 39(4): 28-32. doi: 10.16351/j.1672-6987.2018.04.004. [9] 公衍玲, 郭遥遥, 刘洋青. 樱桃核乙醇提取物体外抗氧化及降糖降脂研究[J]. 青岛科技大学学报(自然科学版),2017,38(6):14−17. [GONG Y L, GUO Y Y, LIU Y Q. Study on antioxidant and hypoglycemic and lipid lowering of cherry kernel ethanol extract in vitro[J]. Journal of Qingdao University of Science and Technology (Natural Science Edition),2017,38(6):14−17. doi: 10.16351/j.1672-6987.2017.06.002[GONG Y L, GUO Y Y, LIU Y Q. Study on antioxidant and hypoglycemic and lipid lowering of cherry kernel ethanol extract in vitro[J]. Journal of Qingdao University of Science and Technology(Natural Science Edition), 2017, 38(6): 14-17. doi: 10.16351/j.1672-6987.2017.06.002. [10] 王竞珮. 甜樱桃果多酚的提取、纯化及其抗氧化、抗哀老活性研究[D]. 太原: 山西农业大学, 2020WANG J P. Extraction and purification of polyphenols of sweet cherry fruit and their antioxidant and anti-aging activities[D]. Taiyuan: Shanxi Agricultural University, 2020. [11] 李雯霞, 于司达, 诸葛斌, 等. 芍药花提取物对果蔬腐败菌的抑菌活性及其在樱桃番茄保鲜中的应用[J]. 食品与发酵工业,2018,44(2):228−232. [LI W X, YU S D, ZHUGE B, et al. Antibacterial activity of peony flower extract on spoilage bacteria in fruits and vegetables and its application in cherry tomato preservation[J]. Food and Fermentation Industry,2018,44(2):228−232. doi: 10.13995/j.cnki.11-1802/ts.015384[LI W X, YU S D, ZHUGE B, et al. Antibacterial activity of peony flower extract on spoilage bacteria in fruits and vegetables and its application in cherry tomato preservation[J]. Food and Fermentation Industry, 2018, 44(2): 228-232. doi: 10.13995/j.cnki.11-1802/ts.015384. [12] 曹小燕, 杨海涛. 超声辅助双水相提取油菜籽粕黄酮及抗氧化性研究[J]. 中国调味品,2019,44(4):52−56, 63. [CAO X Y, YANG H T. Ultrasound-assisted two-phase extraction of flavonoids from rapeseed meal and its antioxidant activity[J]. China Condiments,2019,44(4):52−56, 63.[CAO X Y, YANG H T. Ultrasound-assisted two-phase extraction of flavonoids from rapeseed meal and its antioxidant activity[J]. China Condiments, 2019, 44(4): 52-56;63. [13] YAN J K, MA H L, PEI J J, et al. Facile and effective separation of polysaccharides and proteins from Cordyceps sinensis mycelia by ionic liquid aqueous two-phase system[J]. Separation and Purification Technology,2014,135:278−284. doi: 10.1016/j.seppur.2014.03.020 [14] ALI B. Antioxidant and free radical-scavenging activities of smooth hound (Mustelus mustelus) muscle protein hydrolysates obtained by gastrointestinal proteases[J]. Food Chemistry,2009(114):1198−1205. [15] 杨明琰, 张晓琦, 沈俭, 等. 超氧化物歧化酶两种邻苯三酚自氧化测定活力方法的比较[J]. 微生物学杂志,2006(3):40−42. [YANG M Y, ZHANG X Q, SHEN J, et al. Comparison of two catetriol autooxidation determination methods for superoxide dismutase[J]. Chinese Journal of Microbiology,2006(3):40−42.[YANG M Y, ZHANG X Q, SHEN J, et al. Comparison of two catetriol autooxidation determination methods for superoxide dismutase[J]. Chinese Journal of Microbiology, 2006(3): 40-42. [16] 孔令娜. 阿尔茨海默病中β-淀粉样蛋白寡聚体的光学检测和解聚研究[D]. 上海: 华东师范大学, 2020KONG L N. Optical detection and depolymerization of beta-amyloid oligomers in Alzheimer's disease [D]. Shanghai: East China Normal University, 2020. [17] 朱立猛. 壳寡糖对阿尔茨海默病的作用效果评价及其机制初探[D]. 北京: 中国科学院大学(中国科学院过程工程研究所), 2021: 67−68ZHU L M. Evaluation of the effect of chitosan oligosaccharide on Alzheimer's disease and its mechanism[D]. Beijing: University of Chinese Academy of Sciences (Institute of Process Engineering, Chinese Academy of Sciences), 2021: 67−68. [18] 冯颖. 两种多酚类物质对Aβ42聚集和毒性的影响及Aβ1-16毒性和炎症反应的研究[D]. 沈阳: 中国医科大学, 2010FENG Y. The effects of two polyphenols on the aggregation and toxicity of Aβ42 and the toxicity and inflammatory response of Aβ1-16[D]. Shenyang: China Medical University, 2010. [19] 肖超, 闫啸, 关怡新, 等. 小分子伴侣对淀粉样蛋白β聚集抑制作用研究[J]. 高校化学工程学报,2021,35(2):316−323. [XIAO C, YAN X, GUAN Y X, et al. Inhibitory effect of small molecular chaperones on amyloid β aggregation[J]. Journal of Chemical Engineering in Universities,2021,35(2):316−323.[XIAO C, YAN X, GUAN Y X, et al. Inhibitory effect of small molecular chaperones on amyloid β aggregation[J]. Journal of Chemical Engineering in Universities, 2021, 35(2): 316-323. [20] 王凤, 肖楚翔, 刘淑珍, 等. 榴莲核黄酮的提取及其对秀丽隐杆线虫氧化和衰老的影响[J]. 食品科学,2021,42(9):123−129. [WANG F, XIAO C X, LIU S Z, et al. Extraction of durian riboflavonoids and their effects on oxidation and aging in Caenorhabditis elegans[J]. Food Science,2021,42(9):123−129.[WANG F, XIAO C X, LIU S Z, et al. Extraction of durian riboflavonoids and their effects on oxidation and aging in Caenorhabditis elegans[J]. Food Science, 2021, 42(9): 123-129. [21] 张娇龙. 齐墩果酸对秀丽隐杆线虫寿命的影响及分子机制研究[D]. 天津: 天津大学, 2016: 17−40ZHANG J L. Effects of oleanolic acid on the lifespan of Caenorhabditis elegans and its molecular mechanism[D]. Tianjin: Tianjin University, 2016: 17−40. [22] 吴浩. Src酪氨酸蛋白激酶抑制剂对AD秀丽隐杆线虫Aβ毒性作用的研究[D]. 长春: 吉林大学, 2021WU H. Study on the toxic effect of Src tyrosine protein kinase inhibitor on Aβ of AD Caenorhabditis elegans[D]. Changchun: Jilin University, 2021. [23] 王双慧, 梅群芳, 冯晓汀. 燕麦β-葡聚糖、多酚及黄酮的抑菌活性研究[J]. 食品研究与开发,2020,41(12):96−102. [WANG S H, MEI Q F, FENG X T. Antibacterial activity of oat β-glucans, polyphenols and flavonoids[J]. Food Research and Development,2020,41(12):96−102.[WANG S H, MEI Q F, FENG X T. Antibacterial activity of oat β-glucans, polyphenols and flavonoids[J]. Food Research and Development, 2020, 41(12): 96-102. [24] 高静. 天然抗氧化剂及其协同作用[J]. 食品安全质量检测学报,2020,11(6):1859−1864. [GAO J. Natural antioxidants and their synergistic effects[J]. Journal of Food Safety and Quality Inspection,2020,11(6):1859−1864. doi: 10.19812/j.cnki.jfsq11-5956/ts.2020.06.030[GAO J. Natural antioxidants and their synergistic effects[J]. Journal of Food Safety and Quality Inspection, 2020, 11(6): 1859-1864. doi: 10.19812/j.cnki.jfsq11-5956/ts.2020.06.030. [25] 张翠利, 付丽娜, 杨小云, 等. 活性氧自由基与细胞衰老关系的研究进展[J]. 广州化工,2015,43(19):5−7. [ZHANG C L, FU L N, YANG X Y, et al. Research progress on the relationship between reactive oxygen species and cell aging[J]. Guangzhou Chemical Industry,2015,43(19):5−7. doi: 10.3969/j.issn.1001-9677.2015.19.003[ZHANG C L, FU L N, YANG X Y, et al. Research progress on the relationship between reactive oxygen species and cell aging[J]. Guangzhou Chemical Industry, 2015, 43(19): 5-7. doi: 10.3969/j.issn.1001-9677.2015.19.003 [26] 高帆, 夏惠, 王秀, 等. 不同甜樱桃品种抗氧化物质及抗氧化活性分析[J]. 浙江农业学报,2017,29(6):926−932. [GAO F, XIA H, WANG X, et al. Analysis of antioxidant substances and antioxidant activity in different sweet cherry cultivars[J]. Zhejiang Journal of Agricultural Sciences,2017,29(6):926−932.[GAO F, XIA H, WANG X, et al. Analysis of antioxidant substances and antioxidant activity in different sweet cherry cultivars[J]. Zhejiang Journal of Agricultural Sciences, 2017, 29(6): 926-932. [27] REITER R J, PAREDES S D, KORKMAZ A, et al. Melatonin in relation to the “strong” and “weak”versions of the free radical theory of aging[J]. Advances in Medical Sciences,2008,53:119−129. [28] 李成, 宋吉雪, 刘婷婷, 等. 基于ThT荧光寿命检测蛋清溶菌酶蛋白寡聚体[J]. 高等学校化学学报,2019,40(1):90−95. [LI C, SONG J X, LIU T T, et al. Detection of egg white lysozyme protein oligomers based on ThT fluorescence lifetime[J]. Journal of Chemistry in Universities,2019,40(1):90−95.[LI C, SONG J X, LIU T T, et al. Detection of egg white lysozyme protein oligomers based on ThT fluorescence lifetime[J]. Journal of Chemistry in Universities, 2019, 40(1): 90-95. [29] 刘伟, 孙彦. β-淀粉样蛋白的聚集及其调控[J]. 化工学报,2022,73(6):2381−2396. [LIU W, SUN Y. Aggregation and regulation of β-amyloid protein[J]. CIESC Journal,2022,73(6):2381−2396.[LIU W, SUN Y. Aggregation and regulation of β-amyloid protein[J]. CIESC Journal, 2022, 73(6): 2381-2396. [30] CORSI A K, WIGHTMAN B, CHALFIE M. A transparent window into biology: A primer on Caenorhabditis elegans[J]. Genetics,2015,200(2):387−407. doi: 10.1534/genetics.115.176099 [31] KALETTA T, HENGARTNER M O. Finding function in novel targets: C. elegans as a model organism[J]. Nat Rev Drug Discov,2006,5(5):387−398. doi: 10.1038/nrd2031 [32] GUARENTE L, KENYON C. Genetic pathways that regulate ageing in model organisms[J]. Nature,2000,408(6809):255−262. doi: 10.1038/35041700 [33] BRAECKMAN B P, VANFLETEREN J R. Genetic control of longevity in C. elegans[J]. Experimental Gerontology,2007,42(1−2):90−98. doi: 10.1016/j.exger.2006.04.010 [34] OCKUN M A, GERCEK Y C, DEMIRSOY H, et al. Comparative evaluation of phenolic profile and antioxidant activity of new sweet cherry (Prunus avium L.) genotypes in Turkey[J]. Phytochem Anal,2022,33(4):564−576. doi: 10.1002/pca.3110 [35] WEI H, CHEN X, ZONG X, et al. Comparative transcriptome analysis of genes involved in anthocyanin biosynthesis in the red and yellow fruits of sweet cherry (Prunus avium L.)[J]. Plos One,2015,10(3):e0121164. doi: 10.1371/journal.pone.0121164 [36] DHALARIA R, VERMA R, KUMAR D, et al. Bioactive compounds of edible fruits with their anti-aging properties: A comprehensive review to prolong human life[J]. Antioxidants,2020,9(11):11−23. doi: 10.3390/antiox9111123 [37] WANG B, TANG X, MAO B, et al. Anti-aging effects and mechanisms of anthocyanins and their intestinal microflora metabolites[J]. Critical Reviews in Food Science and Nutrition,2022:1−17. doi: 10.1080/10408398.2022.2123444 [38] JAYARATHNE S, RAMALINGAM L, EDWARDS H, et al. Tart cherry increases lifespan in Caenorhabditis elegans by altering metabolic signaling pathways[J]. Nutrients,2020,12(5):1482. doi: 10.3390/nu12051482 [39] 韩文强, 冯居秦. 樱桃核提取物在中草药化妆品研发中的应用[J]. 中国美容医学,2019,28(9):171−173. [HAN W Q, FENG J Q. Application of cherry pit extract in the research and development of Chinese herbal cosmetics[J]. Chinese Journal of Aesthetic Medicine,2019,28(9):171−173. doi: 10.15909/j.cnki.cn61-1347/r.003292[HAN W Q, FENG J Q. Application of cherry pit extract in the research and development of Chinese herbal cosmetics[J]. Chinese Journal of Aesthetic Medicine, 2019, 28(9): 171-173. doi: 10.15909/j.cnki.cn61-1347/r.003292. [40] 陆晓达. 尼古丁对转基因秀丽隐杆线虫体内Aβ蛋白聚集作用的研究[D]. 长春: 长春理工大学, 2021LU X D. Study on the effect of nicotine on Aβ protein aggregation in transgenic Caenorhabditis elegans[D]. Changchun: Changchun University of Technology, 2021. [41] BOWERS Z, MAITI P, BOURCIER A, et al. Tart cherry extract and omega fatty acids reduce behavioral deficits, gliosis, and amyloid-beta deposition in the 5xFAD mouse model of Alzheimer's disease[J]. Brain Sciences,2021,11(11):1423. doi: 10.3390/brainsci11111423 -
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