香水莲花提取物调节脂代谢作用研究

发布时间：2018-02-20 04:09

  本文关键词： 香水莲花 肥胖 柯里拉京 蜡菊素A 胰脂肪酶 肠道微生物　出处：《浙江大学》2017年硕士论文　论文类型：学位论文

【摘要】：随着生活水平的提高与人们生活方式的改变,肥胖已成为一种公共卫生问题,它的发病率在全球呈不断上升趋势。植物来源的天然产物能通过抑制脂肪吸收、合成,促进脂肪代谢,抑制食欲等方式起到减肥降脂的作用。香水莲花(Nymphaea hybrid)是睡莲科睡莲属热带大型睡莲,富含多酚、黄酮、花色苷等物质,具有多种生物学功效。本文以香水莲花提取物为原料,以胰脂肪酶(pancreatic lipase,PL)抑制率为指针,利用高效制备液相色谱(Preparative High-Performance Liquid Chromatography,Prep-HPLC)技术从香水莲花提取物(Nymphaeahybridextracts,NHE)中分离制备活性单体,并研究了 NHE对小鼠高脂膳食诱导肥胖的预防作用以及对肥胖小鼠肠道微生物的影响。本文研究结果如下:(1)应用Prep-HPLC技术从NHE中分离制备得到十个组分;通过质谱、核磁共振等技术对其中两个PL抑制活性单体进行结构鉴定,分别为β-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-D-glucose(柯里拉京,Corilagin)、4',5,7-Trihydroxyflavanone-5-O-β-D-Glucopyranoside(蜡菊素 A,Helichrysin A)。这两种物质均为酚类物质,均首次在香水莲花中分离得到,它们在NHE中的含量分别为 24.94mg/g、12.34mg/g。(2)优化得到PL抑制实验的最佳反应条件为反应时间10 min、pH 7.5、温度37℃。NHE分级相中香水莲花乙酸乙酯相(NHEA)对PL抑制作用最强,其半抑制浓度(IC50)为2.09mg/mL;从NHE中分离制备得到的十个组分中,六个组分对PL有较强的抑制作用,其中柯里拉京活性最强(IC50为0.64mg/mL),蜡菊素A次之(IC50为0.70 mg/mL)。香水莲花提取物中总酚含量与PL抑制率的相关性高达0.885。(3)采用预防型肥胖小鼠模型,研究NHE对小鼠高脂膳食诱导肥胖的预防作用。结果表明,与模型对照组相比,高剂量(400 mg/kg/BW)NHE能显著降低小鼠体重、附睾周围脂肪重量、肾周围脂肪重量、肝脏重量(p0.05),降低小鼠血清中甘油三酯(TG)、总胆固醇(TC)含量(p0.05),提高粪便中脂类、甘油三酯(TG)含量(p0.05),降低胰腺中PL活性(p0.01),增强肝脏超氧化物歧化酶(SOD)活性(p0.05),降低肝脏丙二醛(MDA)含量(p0.05)以及降低脂肪组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、单核细胞趋化因子-1(MCP-1)含量(p0.01)。(4)采用高通量测序方法,研究NHE对高脂膳食小鼠肠道微生物的影响。结果表明,与模型对照组相比,NHE能增加高脂膳食小鼠肠道微生物的多样性与丰度以及改善菌群结构,其具体表现为降低Firmicutes与Bacteridets的比值(F/B值),增加益生菌Lactobacillius与Akkermansia muciniphila的数量,减少有害菌Desulfovibrio的数量,但无法增加Bifidobacterium的数量。综上所述,香水莲花提取物具有预防肥胖、改善肝脏氧化应激、缓解脂肪组织慢性炎症的作用,并且呈一定的量效关系。推测它能通过抑制胰脂肪酶活性、改善肠道微生态发挥减肥因子的作用,且柯里拉京与蜡菊素A可能为其中的特征化合物,这为香水莲花降脂产品的开发与利用提供了一定的科学依据与理论指导。
[Abstract]:With the improvement of living standards and the change of people's lifestyle, obesity has become a public health problem, its incidence rate showed a rising trend in the world. Natural products from plants can inhibit the absorption of fat, promote fat synthesis, metabolism, appetite suppression way to lose weight and fat role. (lotus fragrance Nymphaea hybrid) is a large tropical water lily Nymphaea lotus, rich in polyphenols, flavonoids, anthocyanins and other substances, has multiple biological functions. In this paper, the perfume lotus extract as raw materials by pancreatic lipase (pancreatic lipase, PL) inhibition rate as the indicator, prepared by high performance liquid chromatography system (Preparative High-Performance Liquid Chromatography, Prep-HPLC) from the perfume lotus extract (Nymphaeahybridextracts, NHE) in the separation and preparation of active monomer, and study the preventive effect of NHE on obesity in mice induced by high fat diet And the influence on the intestinal microflora of obese mice. The results of this study are as follows: (1) the application of Prep-HPLC technology in preparation and separation of ten fractions were obtained from NHE; by mass spectrometry, nuclear magnetic resonance technology in which the two PL inhibitory activity were identified respectively as monomers, beta -1-O-galloyl-3,6- (R) -hexahydroxydiphenoyl-D-glucose (corilagin. Corilagin, 4', 5,7-Trihydroxyflavanone-5-O-) (La Ju A beta -D-Glucopyranoside, Helichrysin A). These two substances are phenolic compounds were isolated for the first time in the lotus fragrance, content in NHE were respectively 24.94mg/g, 12.34mg/g. (2) optimized reaction conditions of PL inhibition for reaction time 10 min, pH 7.5, temperature 37.NHE grade ethyl acetate phase perfume Lotus (NHEA) of PL had the strongest inhibitory effect, the half inhibitory concentration (IC50) was isolated from 2.09mg/mL; preparation of NHE 鍗佷釜缁勫垎涓，

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