铁皮石斛活性成分研究和对皮肤光老化的改善作用

	铁皮石斛活性成分研究和对皮肤光老化的改善作用
	陈定康
导师	胡江苗
关键词	铁皮石斛，超高效液相色谱与四级杆飞行时间质谱，联苄，抗光老化，SIRT3 Dendrobium officinale, UPLC-Q-TOF-MS, Bibenzyl derivatives, Anti-photoaging, SIRT3
摘要	面对老龄化社会，老龄化带来的挑战是一项紧迫的任务。皮肤是我们身体的第一道屏障，保护我们免受感染和脱水。皮肤不仅受到内在老化的影响，还受到外部老化的影响。在日常生活中皮肤的衰老主要因紫外照射诱导表皮细胞产生大量的活性氧引发氧化应激，造成皮肤光老化。ROS介导的Mn-SOD/SIRT3光老化途径，是皮肤光老化的一种重要方式。因此，针对SIRT3这一靶点筛选具有良好抗光老化活性的物质，是未来抗衰药物的研发方向之一。古籍记载，铁皮石斛（Dendrobium officinale）具有延缓皮肤衰老的功效，在现代工业化妆品原料中也常有应用。因此，对铁皮石斛中抗衰老活性物质及其抗衰老机制进行的深入挖掘具有重要的意义，这也是将铁皮石斛作为名贵化妆品原料去推广应用的科学依据。本论文对铁皮石斛的物质构成及其抗光老化机制进行的全面深入的研究。一方面利用超高效液相色谱与四级杆飞行时间质谱（UPLC-Q-TOF-MS），通过定性和定量分析对不同产地铁皮石斛醇提物中小分子构成和含量进行了分析归纳；另一方面建立细胞光老化模型，采取表面等离子体共振技术(Surface Plasmon Resonance, SPR)，Western blot和试剂盒检测等手段，深入探究了铁皮石斛中联苄类化合物与SIRT3蛋白之间的亲和力及其抗光老化的作用机制。主要实验结果有： 1. 通过定性和定量分析，共鉴定了(-)-Lirioresinol B、Fructose-phenylalanine、香草酸、对羟基苯甲酸、石斛碱等32种主要成分，证实了铁皮石斛中富含酚性物质，主要为石斛酚代表的联苄类化合物和柚皮素代表的黄酮类成分； 2. 铁皮石斛中联苄类化合物能以高亲和力与SIRT3直接结合，进而增强SIRT3活性； 3. 铁皮石斛中联苄类化合物改善紫外照射引起的皮肤光老化的作用机制为ROS 介导的Mn-SOD/SIRT3途径。综上所述，铁皮石斛中次生代谢物因生长环境不同虽有一定差异，但其主要活性成分具有很高的共性。特别是以石斛酚和柚皮素为代表的抗衰活性成分，在不同产地均有检测到，只是其含量存在一定差异，这也提示我们可以将这两种成分作为铁皮石斛的潜在质量评价指标。通过实验证实，铁皮石斛中联苄类化合物可以通过ROS介导的Mn-SOD/SIRT3途径改善皮肤的状态。这为铁皮石斛中联苄类化合物的抗光老化作用机制研究提供了理论依据。同时启示我们，铁皮石斛可以作为一种天然皮肤护理产品的植物资源来开发应用。; The challenges presented by aging is an urgent task, as we are facing an aging society. Skin is the first barrier of our body that protects us from infection and dehydration. The skin is influenced not only by intrinsic aging, but also by extrinsic aging, triggered by environmental factors that contribute to accelerating the skin aging process. In daily life, skin aging is mainly due to UV irradiation induced epidermal cells to produce a large amount of reactive oxygen species, which leads to oxidative stress and skin photoaging. ROS mediated Mn-SOD/SIRT3 photoaging pathway is an important way of skin photoaging. Therefore, screening substances against SIRT3 is one of the directions of anti-aging drugs research in the future. According to ancient books, Dendrobium officinale has the effect of delaying skin aging and is often used as modern industrial cosmetic raw materials. It is of great significance to deeply explore the anti-aging active substances and anti-aging mechanism in Dendrobium officinale, which is also the scientific basis for the application of Dendrobium officinale as a valuable cosmetic raw material. In this paper, the material composition and anti-aging mechanism of Dendrobium officinale were studied comprehensively and deeply. On the one hand, using UPLC-Q-TOF-MS technology, the composition and content of small molecules in the alcohol extract of Dendrobium officinale from different producing areas were analyzed and summarized through qualitative and quantitative analysis; On the other hand, the cell photoaging model was established. By means of surface plasmon resonance (SPR), western blot and kit detection, the affinity between benzenes and SIRT3 protein in Dendrobium officinale and its anti-photoaging mechanism were deeply explored. The main experimental results are as follows: 1. Through qualitative and quantitative analysis, 32 main components such as (-) - lirioresinol B, fructose phenylalanine, vanillic acid, p-hydroxybenzoic acid and dendrobine were identified. It was confirmed that Dendrobium officinale was rich in phenolic substances, mainly bibenzyl compounds represented by gigantol and flavones represented by naringenin; 2. Bibenzyl derivatives in Dendrobium officinale can directly bind to SIRT3 with high affinity, and then enhance SIRT3 activity; 3. Bibenzyl derivatives in Dendrobium officinale can enhance the activity of Mn-SOD through SIRT3, and further improve the skin photoaging caused by UV irradiation. In conclusion, the secondary metabolites produced by Dendrobium officinale under the influence of the different growth environment are different, but its main active components have high commonality. Accordingly, the quantitative analysis results showed that there were differences in the contents of gigantol and naringin in Dendrobium officinale from different habitats, indicating that the two compounds can be used as potential quality evaluation indexes of Dendrobium officinale. It is confirmed by experiments t
语种	中文
	2022-05
学位授予单位	中国科学院大学
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/75171
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	陈定康. 铁皮石斛活性成分研究和对皮肤光老化的改善作用[D]. 中国科学院大学,2022.

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