樱桃花色苷的降脂作用及其机制研究

发布时间：2018-04-16 01:03

本文选题：樱桃花色苷 + 抗氧化活性　；参考：《浙江大学》2017年硕士论文

【摘要】：脂质沉积是指肝、胰或者脂肪组织中脂滴数量显著增多且聚集。脂质异位沉积对骨骼肌、胰岛β细胞、肝脏等均有一定的脂毒性作用,并且与糖尿病、脂肪肝、高脂血症等疾病发生密切相关。现有的降脂药物均具有一定的副作用及不良反应。作为药食同源的美味佳果,樱桃富含多种活性营养成分,其中花色苷含量可达4-450mg/100g鲜重。樱桃花色苷有抗氧化、预防心血管疾病以及降脂减肥等功效,可降低胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平,提高载脂蛋白(ApoA)、高密度脂蛋白胆固醇(HDL-C)表达水平。因此,从樱桃中分离制备花色苷并研究其降脂功效具有重要意义。本课题从樱桃中提取樱桃花色苷并对樱桃花色苷的组分及总黄酮、总多酚含量进行测定并分析了樱桃花色苷的抗氧化活性及其对细胞脂质沉积的抑制改善作用,探讨了细胞自噬在樱桃花色苷抑制改善细胞脂质沉积中的作用。课题主要研究结果如下:1.分离纯化所得的樱桃花色苷经HPLC组分鉴定发现其主要成分矢车菊素-3-葡萄糖苷、矢车菊素-3-芸香糖苷、花葵素-3-葡萄糖苷。运用DPPH、ATBS等体外抗氧化实验、细胞实验结合荧光探针对樱桃花色苷的抗氧化活性进行评价分析证实了樱桃花色苷有较强的体外抗氧化能力,且对丙烯酰胺(AA)诱导L02细胞形成的氧化应激模型有较好的保护作用。2.利用油酸(OA)孵育诱导HepG2、L02细胞建立的细胞脂肪沉积模型评价樱桃花色苷对细胞脂质沉积的抑制改善作用,证实樱桃花色苷对HepG2、L02细胞脂质沉积有显著改善作用。樱桃花色苷可显著降低细胞甘油三酯(TG)、总胆固醇(TC)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)含量水平。3.结合透射电镜、免疫荧光等实验进一步探讨细胞自噬在樱桃花色苷抑制改善细胞脂质沉积中的作用,发现樱桃花色苷可通过促进细胞自噬体形成,上调自噬相关蛋白LC3、Atg5、Beclin1表达水平,下调P62的表达水平进一步缓解抑制HepG2、L02脂质沉积作用,樱桃花色苷可下调脂肪酸合成相关蛋白FAS、ACC的表达,从而证实了细胞自噬参与樱桃花色苷的降脂作用。
[Abstract]:Lipid deposition refers to the significant increase and accumulation of lipid droplets in liver, pancreas or adipose tissue.Lipid ectopic deposition has some lipid toxicity on skeletal muscle, islet 尾 cells and liver, and is closely related to diabetes mellitus, fatty liver, hyperlipidemia and other diseases.The existing lipid-lowering drugs have some side effects and adverse reactions.As a delicious fruit, cherry is rich in many kinds of active nutrients, among which anthocyanin content can reach 4-450mg/100g fresh weight.Cherry anthocyanin has the functions of anti-oxidation, preventing cardiovascular disease and reducing fat and weight loss. It can reduce the levels of TC, TG, LDL-C, and increase the expression of ApoAn and HDL-C.Therefore, it is of great significance to isolate anthocyanin from cherry and study its lipid-lowering effect.In this study, cherry anthocyanin was extracted from cherry and its components, total flavonoids and total polyphenols were determined. The antioxidant activity of cherry anthocyanin and its effect on the inhibition of cellular lipid deposition were analyzed.The role of autophagy in the inhibition of cherry anthocyanin in improving cell lipid deposition was investigated.The main research results are as follows: 1: 1.The main components of the purified Cherry anthocyanin were identified by HPLC. The main components were cornulin-3-glucoside, cornulin-3-rutinoside and anthocyanin-3-glucoside.The antioxidant activity of cherry anthocyanin was evaluated by using DPPH ATBS and fluorescence probe in vitro. The results showed that cherry anthocyanin had strong antioxidant ability in vitro.And it has a good protective effect on the oxidative stress model of L02 cells induced by acrylamide (AA). 2.The effect of cherry anthocyanin on the inhibition of lipid deposition in HepG2L02 cells was evaluated by using oleic acid (OAA) incubation model. It was proved that cherry anthocyanin could significantly improve the lipid deposition of HepG2L02 cells.Cherry anthocyanin could significantly reduce the contents of TGG, TCU, alt, AST3 in the cell lines of triglyceride, total cholesterol, alanine aminotransferase and aspartate aminotransferase (AST).In combination with transmission electron microscopy and immunofluorescence assay, the role of cell autophagy in the inhibition of cherry anthocyanin in improving cell lipid deposition was further investigated. It was found that cherry anthocyanin could promote the formation of autophagy.The up-regulated expression of autophagy associated protein LC3 / Atg5 / Beclin1, down-regulated the expression of P62 and further alleviated the inhibition of lipid deposition in HepG2 / L02. Cherry anthocyanin could down-regulate the expression of fatty acid synthase associated protein FASA ACC.It was proved that autophagy was involved in the lipid-lowering effect of Cherry anthocyanin.
【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】：R285.5

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