本文關(guān)鍵詞： 香水蓮花 肥胖 柯里拉京 蠟菊素A 胰脂肪酶 腸道微生物　出處：《浙江大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：隨著生活水平的提高與人們生活方式的改變,肥胖已成為一種公共衛(wèi)生問題,它的發(fā)病率在全球呈不斷上升趨勢。植物來源的天然產(chǎn)物能通過抑制脂肪吸收、合成,促進(jìn)脂肪代謝,抑制食欲等方式起到減肥降脂的作用。香水蓮花(Nymphaea hybrid)是睡蓮科睡蓮屬熱帶大型睡蓮,富含多酚、黃酮、花色苷等物質(zhì),具有多種生物學(xué)功效。本文以香水蓮花提取物為原料,以胰脂肪酶(pancreatic lipase,PL)抑制率為指針,利用高效制備液相色譜(Preparative High-Performance Liquid Chromatography,Prep-HPLC)技術(shù)從香水蓮花提取物(Nymphaeahybridextracts,NHE)中分離制備活性單體,并研究了 NHE對(duì)小鼠高脂膳食誘導(dǎo)肥胖的預(yù)防作用以及對(duì)肥胖小鼠腸道微生物的影響。本文研究結(jié)果如下:(1)應(yīng)用Prep-HPLC技術(shù)從NHE中分離制備得到十個(gè)組分;通過質(zhì)譜、核磁共振等技術(shù)對(duì)其中兩個(gè)PL抑制活性單體進(jìn)行結(jié)構(gòu)鑒定,分別為β-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-D-glucose(柯里拉京,Corilagin)、4',5,7-Trihydroxyflavanone-5-O-β-D-Glucopyranoside(蠟菊素 A,Helichrysin A)。這兩種物質(zhì)均為酚類物質(zhì),均首次在香水蓮花中分離得到,它們?cè)贜HE中的含量分別為 24.94mg/g、12.34mg/g。(2)優(yōu)化得到PL抑制實(shí)驗(yàn)的最佳反應(yīng)條件為反應(yīng)時(shí)間10 min、pH 7.5、溫度37℃。NHE分級(jí)相中香水蓮花乙酸乙酯相(NHEA)對(duì)PL抑制作用最強(qiáng),其半抑制濃度(IC50)為2.09mg/mL;從NHE中分離制備得到的十個(gè)組分中,六個(gè)組分對(duì)PL有較強(qiáng)的抑制作用,其中柯里拉京活性最強(qiáng)(IC50為0.64mg/mL),蠟菊素A次之(IC50為0.70 mg/mL)。香水蓮花提取物中總酚含量與PL抑制率的相關(guān)性高達(dá)0.885。(3)采用預(yù)防型肥胖小鼠模型,研究NHE對(duì)小鼠高脂膳食誘導(dǎo)肥胖的預(yù)防作用。結(jié)果表明,與模型對(duì)照組相比,高劑量(400 mg/kg/BW)NHE能顯著降低小鼠體重、附睪周圍脂肪重量、腎周圍脂肪重量、肝臟重量(p0.05),降低小鼠血清中甘油三酯(TG)、總膽固醇(TC)含量(p0.05),提高糞便中脂類、甘油三酯(TG)含量(p0.05),降低胰腺中PL活性(p0.01),增強(qiáng)肝臟超氧化物歧化酶(SOD)活性(p0.05),降低肝臟丙二醛(MDA)含量(p0.05)以及降低脂肪組織中腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素-6(IL-6)、單核細(xì)胞趨化因子-1(MCP-1)含量(p0.01)。(4)采用高通量測序方法,研究NHE對(duì)高脂膳食小鼠腸道微生物的影響。結(jié)果表明,與模型對(duì)照組相比,NHE能增加高脂膳食小鼠腸道微生物的多樣性與豐度以及改善菌群結(jié)構(gòu),其具體表現(xiàn)為降低Firmicutes與Bacteridets的比值(F/B值),增加益生菌Lactobacillius與Akkermansia muciniphila的數(shù)量,減少有害菌Desulfovibrio的數(shù)量,但無法增加Bifidobacterium的數(shù)量。綜上所述,香水蓮花提取物具有預(yù)防肥胖、改善肝臟氧化應(yīng)激、緩解脂肪組織慢性炎癥的作用,并且呈一定的量效關(guān)系。推測它能通過抑制胰脂肪酶活性、改善腸道微生態(tài)發(fā)揮減肥因子的作用,且柯里拉京與蠟菊素A可能為其中的特征化合物,這為香水蓮花降脂產(chǎn)品的開發(fā)與利用提供了一定的科學(xué)依據(jù)與理論指導(dǎo)。
[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 鍗佷釜緇勫垎涓，

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