【作者】 王慧；

【导师】 赵江； 刘丹；

【作者基本信息】 天津科技大学 ， 食品工程（专业学位）， 2019， 硕士

【摘要】 D-手性肌醇是一种羟基环己烷,广泛存在于自然界中的荞麦、大豆中,具有良好的植物来源。D-松醇是D-手性肌醇的甲基化衍生物,也是一种在自然界中普遍存在的物质。两者都具有调节血糖浓度、抗氧化、抗炎、抗癌等多种生物功能,但目前对两者的活性研究主要集中在降血糖方面。本论文以果蝇与线虫为实验模式生物,对D-手性肌醇和D-松醇延缓衰老作用进行了研究,并对其作用的分子机制进行初步探讨,为深入探究D-手性肌醇和D-松醇延缓衰老作用的机制提供基础,为相关产品的开发应用提供一定的理论依据。实验以0、10、30 μmol/L的D-手性肌醇和D-松醇饲喂黑腹果蝇,以0、5、10μmol/L的D-手性肌醇和D-松醇饲喂秀丽隐杆线虫。在以黑腹果蝇为模式生物的实验中,分别从其寿命、爬行能力、肠道稳态、延缓衰老相关酶活力以及相关基因mRNA表达水平等方面来研究D-手性肌醇和D-松醇延缓果蝇衰老的作用,且进一步建立双氧水、百草枯急性损伤模型来研究D-手性肌醇和D-松醇对果蝇的急性应激耐受能力的影响。在以秀丽隐杆线虫为模式生物的实验中,分别从其寿命、脂褐素水平、ROS水平、延缓衰老相关酶活力以及相关基因mRNA表达水平等方面来研究D-手性肌醇和D-松醇延缓线虫衰老的作用,并且建立线虫高糖慢性损伤模型,双氧水、百草枯急性损伤模型来研究D-手性肌醇和D-松醇对线虫的保护作用及应激耐受能力的影响。以果蝇为生物模型的实验研究发现,D-手性肌醇和D-松醇能够显著延长果蝇的寿命,提高果蝇的爬行能力,促进果蝇肠道稳态平衡,增强抗氧化酶活力,抑制胰岛素信号通路转导过程,上调相关抗衰老基因的表达水平。在双氧水和百草枯急性氧化损伤实验中,D-手性肌醇和D-松醇能够延长氧化损伤果蝇的寿命,对急性损伤果蝇具有一定的保护作用。在以线虫为生物模型的实验中发现,D-手性肌醇和D-松醇能显著延长线虫的寿命,降低线虫体内脂褐素含量和ROS水平,增加体内抗氧化酶活力以及抑制与线虫衰老相关的信号通路转导过程,显著影响此通路中的基因表达水平。在高糖慢性损伤实验中,D-手性肌醇和D-松醇能显著延长高糖损伤线虫的寿命,降低线虫体内的氧化损伤水平。在线虫的双氧水、百草枯急性损伤实验中,D-手性肌醇和D-松醇实验组的线虫寿命均有效延长,荧光显微镜观察发现D-手性肌醇和D-松醇干预后线虫体内脂褐素含量降低。研究结果表明,D-手性肌醇和D-松醇具有延缓衰老的作用,可以作为相关保健食品的原料。更多还原

【Abstract】 D-Chiro-Inositol(DCI)is a kind of hydroxyl cyclohexane and widely exists in buckwheat and soybean in nature.D-pinitol(DP),one of methylized derivatives of DCI,is also a substance widely existing in nature.Both of them have possessed multiple biological functions such as regulating glucose metabolism antioxidant,anti-inflammatory,anticancer etc.In this paper,we investigated the blood glucose-lowering effect of DCI and DP.Drosophila melanogaster and Caenorhabditis elegans were choosed as experimental model organisms in order to evaluate anti-aging activity of DCI and DP.The preliminary study of the molecular mechanism of its action and further antioxidant mechanisms of DCI and DP were also been explored,which will provide some guidance for the development of related products.The drosophila and nematodes were fed with DCI and DP at different concentrations(Drosophila melanogaster 0、10、30μmol/L,Caenorhabditis elegans 0、5、10 μmol/L)Lifespan,climbing abilities,the intestinal microecological balance,the antioxidant enzyme activity,and anti-aging related genes mRNA expression levels were determined to study anti-aging effect of DCI and DP in the Drosophila melanogaster model organisms.And a hydrogen peroxide,paraquat damage model was further established to study on acute stress tolerance ability of DCI and DP on fruit flies.Lifespan,fluorescence content of lipofuscin,fluorescence content of ROS,the antioxidant enzyme activity,and anti-aging related genes mRNA expression levels were also detected in the Caenorhabditis elegans model organisms.And a high-glucose,hydrogen peroxide,paraquat damage model was further established to study the effect of DCI and DP on stress tolerance ability of C.elegans.It was shown that DCI and DP could significantly prolong the lifespan of drosophila,improve crawling ability,promote the steady state balance of intestinal,enhance the activity of antioxidant enzymes,inhibit the insulin signaling and up-regulated relevant the oxidation resistance related gene mRNA expression levels.In addition,it also had a effect on attenuating the injury induced by paraquat and hydrogen peroxide in fruit flies.It was also shown that the lifespan of C.elegans was prolonged obviously by DCI and DP addition.The antioxidant enzymes activity of SOD increased with the regulation of the expression levels of genes related to aging.The results showed that the C.elegans treated by DCI and DP had the longer survival time compared to that of the control group.Besides DCI and DP could significantly decreased lipofuscin’s autofluorescence and ROS level in vivo of C.elegans with a dose-dependent manner.DCI and DP possessing the capacity of antioxidant and senescence-resistance could be used as raw materials for health food.更多还原