富含A型原花青素的蔓越莓浓缩物体内延缓衰老功能研究

发布时间：2018-05-17 18:57

本文选题：蔓越莓浓缩物 + 葡萄籽提取物　；参考：《浙江大学》2017年硕士论文

【摘要】：蔓越莓(Cranberry)作为一种低热量、高纤维、富含多种营养物质的水果,被誉为北美三大传统水果之一,其制品也正逐渐被引入中国市场。由于蔓越莓中含有大量植物活性物质,尤其是其独特的A型原花青素,能有效保持泌尿道的健康,近年来越来越受到人们的关注。本文以富含A型原花青素的蔓越莓浓缩物(CBC)为研究对象,以富含B型原花青素的葡萄籽提取物(GSE)作为对照品,对其进行了有关主要成分的分析鉴定、小鼠体内延缓衰老和调节氧化还原状态的功能,以及端粒保护作用的研究。得出以原花青素A2为主要生物活性物质的CBC能从调节体内氧化还原状态和端粒保护两方面发挥延缓衰老的作用,相比以(表)儿茶素和原花青素B2为主要生物活性物质的GSE,CBC在谷胱甘肽过氧化物酶(GSH-Px)活性的调节、端粒保护方面的作用效果更加理想,提示CBC作为一种延缓衰老的功能性食品具有巨大潜力。详细研究内容和结果如下:1、主要成分含量分析采用GB法对CBC和GSE中的水分、灰分、脂肪、蛋白质、总黄酮、总原花青素、抗坏血酸和维生素E(以α-生育酚计)的含量进行检测。结果显示CBC与GSE中最主要的活性物质均为原花青素。CBC中原花青素含量为15.6g/kg,GSE中原花青素含量为309.0 g/kg。2、原花青素的组成分析与结构鉴定采用Q-Extractive组合型四极杆/轨道阱高分辨质谱法(QE)对CBC和GSE原花青素的组成(即不同聚合度的原花青素)成分进行分析,结果显示CBC中最主要的酚类活性物质为原花青素A2,其次为(表)儿茶素;而GSE中最主要的酚类活性物质为(表)儿茶素和原花青素B2。3、体内氧化还原状态调节与延缓衰老作用采用D-半乳糖致衰小鼠模型,以水溶性维生素E作为阳性对照,连续8周将15 mg/kg/d、30 mg/kg/d、60 mg/kg/d(剂量以总原花青素含量计)的CBC和GSE分别对受试小鼠进行灌胃。结果显示CBC和GSE可使肝脏丙二醛(MDA,以硫代巴比妥反应物TBARS的量表示)含量降低30%-57%,使大脑MDA含量降低24%-30%,使血浆8-异前列腺素含量降低11%-62%。同时CBC和GSE可使大脑的单胺氧化酶(MAO)活力降低27%-59%,使血浆MAO活力降低65%-71%。高剂量组(60 mg/kg/d)的CBC可使肝脏的GSH-Px水平显著提高42%(p0.05),而中剂量组(30mg/kg/d)的GSE则使肝脏的超氧化物歧化酶(SOD)水平提高了 13%(p0.05)。以上结果表明,CBC和GSE均能通过调节体内氧化还原状态而延缓衰老。但是CBC对肝脏和血清中GSH-Px活性调节效果明显,而GSE对GSH-Px活性无调节作用,而两者对过氧化氢酶(CAT)的活性均无显著影响。4、基于端粒保护的延缓衰老作用采用实时荧光定量聚合酶链反应(RT-PCR)法测定各组小鼠肝脏和睾丸的相对端粒长度(TL),发现低剂量组的CBC能显著抑制肝脏和睾丸中因D-半乳糖致衰而导致的端粒长度缩短(p0.05),从而起到有效保护端粒的作用,但GSE却没有这方面的作用效果。采用蛋白质印迹法(WesternBlot)进行端粒酶逆转录酶基因tert和长寿基因sirt6表达水平的检测,发现中、高剂量组的CBC和低剂量组GSE均能显著降低睾丸中tert蛋白表达水平(p0.05),但只有低剂量组的CBC能显著降低肝脏中的tert蛋白表达水平(p0.05),提示CBC和GSE均可通过降低端粒酶活性从而预防细胞癌变的效果,但作用靶点不同;对于长寿基因sirt6,各剂量组的CBC和中剂量组的GSE均可显著提高长寿基因sirt6的蛋白表达水平(p0.05)。由此可见,CBC和GSE均可靶向性地调控端粒酶逆转录酶基因terr的蛋白表达水平,促进长寿基因sirt6的蛋白表达,但只有CBC起到有效保护端粒的效果。
[Abstract]:Cranberry (Cranberry), as a low calorie, high fiber, and rich variety of nutrients, is known as one of the three major traditional fruits in North America. Its products are also being introduced into the Chinese market. Because the cranberry contains a large number of plant active substances, especially its unique A proanthocyanidins, it can effectively maintain the health of the urinary tract in recent years. More and more people pay attention to it. In this paper, the cranberry concentrate (CBC) rich in A proanthocyanidins (CBC) is used as the research object. The grape seed extract (GSE), which is rich in B proanthocyanidins, is used as a control product. The analysis and identification of the main components, the function of retarding aging and the regulation of redox state in mice, and the telomere protection in mice are carried out in this paper. It is concluded that CBC with proanthocyanidin A2 as the main bioactive substance can play a role in delaying senescence from two aspects of regulating the redox state of the body and the telomere protection, compared with the GSE of (surface) catechin and proanthocyanidin B2 as the main bioactive substance, the regulation of CBC in the activity of cereal caspase peroxidase (GSH-Px), and the telomere. The protective effect is more ideal, suggesting that CBC has great potential as a functional food for delaying senility. The detailed content and results are as follows: 1, the main component content analysis uses GB method for water, ash, fat, protein, total flavonoids, total anthocyanins, ascorbic acid and vitamin E (alpha tocopherol) in CBC and GSE. The results showed that the main active substances in CBC and GSE were proanthocyanidin.CBC and proanthocyanidin content 15.6g/kg, GSE proanthocyanidin content was 309 g/kg.2, and the composition analysis and structural identification of proanthocyanidins were identified by Q-Extractive combined quadrupole / track well high resolution mass spectrometry (QE) for CBC and GSE original flowers. The composition of penicillin (the proanthocyanidins of different degrees of polymerization) was analyzed. The results showed that the main phenolic active substance in CBC was proanthocyanidin A2, followed by (surface) catechin, while the most important phenolic active substance in GSE was (surface) catechin and proanthocyanidin B2.3. The regulation of redox state in the body and the anti aging effect of D- half were used in D- half. The model of lactose senility mice was treated with water soluble vitamin E as positive control. 15 mg/kg/d, 30 mg/kg/d, 60 mg/kg/d (CBC and GSE) were administered to the mice for 8 weeks. The results showed that CBC and GSE reduced the content of the liver malondialdehyde (MDA, the amount of TBARS of thiobarbitone reaction) by 30%. -57%, the content of MDA in the brain decreased 24%-30%, the plasma 8- ISO prostaglandin content decreased 11%-62%. and CBC and GSE could reduce the activity of monoamine oxidase (MAO) activity in the brain 27%-59%, and reduce the activity of MAO activity in the 65%-71%. high dose group (60 mg/kg/d). The superoxide dismutase (SOD) level of the liver was increased by 13% (P0.05). The results showed that both CBC and GSE could delay senescence by regulating the redox state in the body, but CBC had a significant effect on the activity of GSH-Px in the liver and serum, while GSE had no regulation on the activity of GSH-Px, and both of them had no activity on the activity of catalase (CAT). .4, based on the delayed senescence of telomere protection, the relative telomere length (TL) of liver and testis of mice in each group was measured by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). It was found that the CBC of low dose group could significantly inhibit the telomere length shortened (P0.05) caused by D- galactose induced failure in the liver and testis (P0.05), thus it was effective. The effect of telomere was protected, but GSE did not effect this effect. The expression level of telomerase reverse transcriptase gene TERT and longevity gene SIRT6 was detected by Western blot method (WesternBlot). It was found that CBC and low dose group GSE in high dose group could significantly reduce the expression level of TERT protein (P0.05) in the testosterone pill, but only low dose. CBC can significantly reduce the level of TERT protein expression in the liver (P0.05), suggesting that both CBC and GSE can reduce telomerase activity to prevent cell carcinogenesis, but the target is different. For long life gene SIRT6, GSE of each dose group CBC and medium dose group can significantly improve the protein expression level of longevity gene SIRT6 (P0.05). It can be seen that both CBC and GSE can regulate the protein expression level of telomerase reverse transcriptase gene terr and promote the protein expression of long life gene SIRT6, but only CBC can effectively protect telomere.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS255.1

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