桂花提取物的抗氧化及延缓衰老作用研究

发布时间：2018-09-06 14:54

【摘要】：衰老是生物进程的必然阶段,同时伴随各种增龄性和退行性疾病,如心脑血管疾病、糖尿病、呼吸系统疾病、癌症等。延缓衰老,提高生命质量,具有重要的意义。桂花在我国有悠久的食用历史,国内外关于桂花的研究主要集中在香气成分的分析、桂花品种资源整理上,对桂花的药理作用研究较少。本研究以金桂花为原料,以果蝇为实验动物,对桂花提取物的体外抗氧化、体内抗氧化、延缓衰老的作用进行评价,旨在研究桂花的抗氧化和延缓衰老作用及机理,为桂花作为保健食品的研究与开发提供实验理论依据。测定16种常见可食花卉的水提液的总多酚与总黄酮含量及其抗氧化作用。Folin-Ciocalteu法测定总多酚含量,NaNO2-AlCl3-NaOH法测定总黄酮含量,FRAP、DPPH·、ABTS+·法测定抗氧化作用。在16种可食花卉水提液中,总多酚含量以牡丹花、紫玫瑰、桂花最多,红巧梅、茉莉花、金盏菊最少;总黄酮含量以桂花、薰衣草、金银花最多,红巧梅、茉莉花、洛神花最少;抗氧化作用以牡丹花、紫玫瑰、月季、桂花最强,红巧梅、茉莉花、金盏菊最弱。采用自然阴干的金桂花,以无水乙醇作为提取溶剂,在50℃条件下多次回流提取桂花中的有效成分,经过抽滤、旋转蒸发(40℃,-0.1MPa)、真空干燥(40℃,-0.1MPa)、低温冷藏(-4℃)步骤,制得棕黄色软浸膏,1g浸膏相当于2g桂花的有效成分。对浸膏进行基本成分分析,发现其总多糖含量为307.98土8.90mgDE/g,对浸膏进行体外抗氧化作用测定,发现其总黄酮含量为482.27±6.19mgQE/g,总多酚含量为126.73±1.19mgGAE/g。其清除FRAP能力相当于 108.54±0.35mgVC/g,清除 DPPH 能力相当于 65.09±0.75mgVC/g,清除ABTS+.能力相当于 123.23±1.09mgVC/g。以果蝇为实验动物,在其培养基中分别添加不同剂量(0,2.5mg/mL,5mg/mL,10mg/mL,15mg/mL)的桂花提取物,研究桂花提取物对♂和♀果蝇的平均寿命、最高寿命的影响。与对照组相比,添加桂花提取物各剂量组的♂和早果蝇的平均寿命、最高寿命均有所延长,且存在剂量效应关系。在第25d龄和45d龄时,处死果蝇,测定果蝇体内抗氧化物酶活性和过氧化脂质含量,与对照组相比,添加桂花提取物各剂量组的♂和♀果蝇体内的T-SOD、Cu，Zn-SOD、Mn-SOD、CAT活性较对照组均有所提高,且存在剂量效应关系;与对照组相比,添加桂花提取物各剂量组的♂和早果蝇体内MDA含量均有所下降,且存在剂量效应关系。以果蝇为实验动物,在其培养基中分别添加不同剂量(0,2.5mg/mL,5mg/mL,10mg/mL,15mg/mL)的桂花提取物,在第30d龄时进行应激(·OH急性损伤、SDS急性损伤、热应激损伤)试验,研究桂花提取物对果蝇应激试验的作用。与对照组相比,添加桂花提取物各剂量组的♂和早果蝇在各应激试验中的寿命较均有所延长,且存在剂量效应关系。桂花提取物有丰富的总多酚、总黄酮、总多糖含量和较好的体内外抗氧化作用。在动物实验中,能显著提高果蝇的寿命和内源性抗氧化酶体系的活性,降低脂质过氧化物的含量,显著提高果蝇的应激能力。因此,桂花提取物具有较好的体内外抗氧化和延缓衰老的作用。
[Abstract]:Aging is an inevitable stage of biological process, accompanied by a variety of aging and degenerative diseases, such as cardiovascular and cerebrovascular diseases, diabetes, respiratory diseases, cancer and so on. In this study, Osmanthus fragrans was used as raw material and Drosophila melanogaster was used as experimental animal to evaluate the anti-oxidation in vitro, anti-oxidation in vivo and anti-aging effects of Osmanthus fragrans extracts, aiming to study the anti-oxidation and anti-aging effects and mechanism of Osmanthus fragrans, and to provide health care for Osmanthus fragrans. The contents of total polyphenols and flavonoids in water extracts of 16 kinds of common edible flowers and their antioxidant activities were determined. The contents of total polyphenols were determined by Folin-Ciocalteu method, the contents of total flavonoids were determined by NaNO2-AlCl3-NaOH method, the contents of total flavonoids were determined by FRAP, DPPH and ABTS+. The antioxidant activities were determined by ABTS+. Peony, purple rose, osmanthus fragrans, red Qiaomei, jasmine and Calendula are the most abundant in total polyphenols; osmanthus fragrans, lavender, honeysuckle, red Qiaomei, jasmine and Luoshen are the most abundant in total flavonoids; peony, purple rose, rose, rose, rose, osmanthus are the strongest, red Qiaomei, jasmine and Calendula are the weakest in antioxidant activity. The effective components in Osmanthus fragrans were extracted by reflux at 50 C for several times. After filtration, rotary evaporation (40 C, - 0.1MPa), vacuum drying (40 C, - 0.1MPa) and cryopreservation (- 4 C), the brown yellow ointment was prepared. 1g extract was equivalent to 2G of Osmanthus fragrans. Total polysaccharide content was 307.98 soil 8.90 mg DE/g. The total flavonoids content was 482.27 (+6.19 mg QE/g) and total polyphenols content was 126.73 (+1.19 mg GAE/g). The FRAP scavenging capacity was 108.54 (+0.35 mg VC/g), DPPH scavenging capacity was 65.09 (+0.75 mg VC/g) and ABTS +. MgVC/g. Drosophila melanogaster was used as experimental animal. Osmanthus fragrans extract of different doses (0,2.5mg/mL, 5mg/mL, 10mg/mL, 15mg/mL) was added to the medium to study the effect of Osmanthus fragrans extract on the average life span and maximum life span of and Drosophila melanogaster. Compared with the control group, the average life span of and early Drosophila melanogaster were the highest. The antioxidant enzymes activity and lipid peroxide content in Drosophila melanogaster were measured at 25 and 45 days of age. Compared with the control group, the T-SOD, Cu, Zn-SOD, Mn-SOD and CAT activities in the groups with different doses of Osmanthus fragrans extract were higher than those in the control group. Compared with the control group, the content of MDA in each dose group decreased, and there was a dose-effect relationship. Drosophila melanogaster was used as the experimental animal, and different doses (0,2.5mg/mL, 5mg/mL, 10mg/mL, 15mg/mL) of Osmanthus fragrans extract were added to the culture medium for stress at the 30th day of age. (OH acute injury, SDS acute injury, heat stress injury) test, to study the effect of Osmanthus fragrans extract on stress test of drosophila. Compared with the control group, the life span of and Drosophila melanogaster in different doses of Osmanthus fragrans extract groups were prolonged, and there was a dose-effect relationship. Osmanthus fragrans extract was rich in total polyphenols, total yellow. Ketones, total polysaccharides and better antioxidant activities in vivo and in vitro. In animal experiments, Osmanthus fragrans extracts can significantly improve the longevity of Drosophila melanogaster and the activity of endogenous antioxidant enzymes, reduce the content of lipid peroxides, and significantly improve the stress capacity of Drosophila melanogaster.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R151

【参考文献】