本文選題：荔枝蜜 + 棗花蜜　； 參考：《西北大學(xué)》2017年碩士論文

【摘要】：荔枝蜜和棗花蜜是我國(guó)市場(chǎng)上較為常見的兩大單花種蜂蜜。棗花蜜多產(chǎn)自我國(guó)北方地區(qū),而荔枝蜜多產(chǎn)自南方地區(qū)。雖蜂蜜采自不同的植物源,色澤口感各有特色,但都具有豐富的營(yíng)養(yǎng)價(jià)值。然而,目前對(duì)荔枝蜜和棗花蜜的研究卻有所不足,對(duì)花源標(biāo)識(shí)物的研究以及在細(xì)胞水平對(duì)有害細(xì)胞的作用均鮮有報(bào)道。鑒別蜂蜜花源標(biāo)識(shí)物是判別蜂蜜真假、區(qū)分摻假蜂蜜的一種重要手段。而細(xì)胞活性研究則對(duì)了解蜂蜜的藥用價(jià)值以及功能開發(fā)具有重要意義。因此,本課題圍繞荔枝蜜的花源標(biāo)識(shí)物和棗花蜜對(duì)HepG2人肝癌細(xì)胞的促凋亡作用進(jìn)行初步探究。1、構(gòu)建了采自中國(guó)不同地區(qū)的13批荔枝蜜和13批棗花蜜的HPLC-DAD指紋圖譜,獲取相關(guān)系數(shù)矩陣數(shù)據(jù),并對(duì)樣品進(jìn)行聚類分析。指紋圖譜結(jié)果顯示,荔枝蜜共有28個(gè)共有峰,棗花蜜共有32個(gè)共有峰。于荔枝蜜中發(fā)現(xiàn)2種化合物,棗花蜜中發(fā)現(xiàn)4種化合物可作為其花源標(biāo)識(shí)物。聚類分析結(jié)果表明基于相關(guān)系數(shù)矩陣進(jìn)行最遠(yuǎn)距離聚類可在一定程度上分析樣品的地理源,為地理源差異研究提供支持。2、對(duì)荔枝蜜中發(fā)現(xiàn)的2種花源標(biāo)識(shí)物(B1和B2)進(jìn)行分離純化和結(jié)構(gòu)鑒定。采用XAD-2吸附樹脂和半制備型液相色譜分離,分別得到兩種花源標(biāo)識(shí)物22.5 mg和4.7 mg,經(jīng)Q/TOF-MS驗(yàn)證,其分子量均為264,是脫落酸及其同分異構(gòu)體。3、對(duì)荔枝蜜和棗花蜜的體外抗氧化活性進(jìn)行研究,分別測(cè)定了總酚含量、DPPH·清除能力和氧自由基清除能力(ORAC)。結(jié)果顯示,棗花蜜的總酚含量高于荔枝蜜,均值為525.1±72.35 mg GAE/kg。棗花蜜DPPH·清除率(IC50)均值和ORAC均值為32.55±4.16 mg/mL和3.39±0.85 μmol TE/g,明顯高于荔枝蜜,且與總酚含量表現(xiàn)出正相關(guān)。因此,選擇棗花蜜為研究對(duì)象進(jìn)行下一步研究。4、研究了棗花蜜對(duì)HepG2人肝癌細(xì)胞的促凋亡作用,并探究了其誘導(dǎo)凋亡的機(jī)制。結(jié)果顯示,經(jīng)棗花蜜處理的HepG2細(xì)胞活力會(huì)隨著棗花蜜濃度的增加而下降。棗花蜜可以引發(fā)HepG2細(xì)胞凋亡,使其細(xì)胞核發(fā)生破裂,凋亡特征明顯,且凋亡細(xì)胞比例與棗花蜜的濃度也表現(xiàn)出濃度正相關(guān)性。此外,棗花蜜會(huì)引起線粒體膜電位降低、增加Bax和Bad蛋白的表達(dá)量,降低Bcl-2和Bcl-xL蛋白的表達(dá)量。同時(shí),抑癌基因p53的表達(dá)量也伴隨處理濃度增大而增加,caspase-9和下游蛋白caspase-3表達(dá)上調(diào),最終導(dǎo)致凋亡。可見,棗花蜜可能通過p53介導(dǎo)線粒體依賴途徑誘導(dǎo)HepG2細(xì)胞凋亡。
[Abstract]:Litchi honey and jujube nectar are two common single-flower honey in Chinese market. Jujube nectar comes from the north of China, while litchi honey from the south. Honey from different plant sources, color and taste have their own characteristics, but have rich nutritional value. However, the researches on litchi honey and jujube nectar are not enough, and there are few reports on the floral markers and the effects on harmful cells at the cell level. The identification of honey flower source marks is an important means to distinguish the true and false honey and to distinguish the adulteration of honey. The study of cell activity is of great significance to understand the medicinal value and function development of honey. Therefore, the effects of flower marker of litchi honey and jujube nectar on apoptosis of HepG2 human hepatoma cells were studied in this paper. The HPLC-DAD fingerprints of 13 batches of litchi honey and 13 batches of jujube nectar collected from different regions of China were constructed. The correlation coefficient matrix data were obtained and cluster analysis was carried out on the samples. The fingerprint showed that there were 28 peaks in litchi honey and 32 peaks in jujube nectar. Two compounds were found in litchi honey and four compounds in jujube nectar. The results of cluster analysis show that the remote clustering based on the correlation coefficient matrix can analyze the geographical source of the sample to a certain extent. To provide support for the study of geographical source difference. 2. To isolate and purify and identify the structure of two floral markers (B _ 1 and B _ 2) found in litchi honey. Two flower markers, 22.5 mg and 4.7 mg, were obtained by XAD-2 adsorption resin and semi-preparation liquid chromatography, respectively, and were verified by Q/TOF-MS. Its molecular weight is 264, which is abscisic acid and its isomer. 3. The antioxidant activities of litchi honey and jujube nectar in vitro were studied. The scavenging capacity of DPPH and oxygen free radical were determined respectively. The results showed that the total phenol content of jujube nectar was higher than that of litchi honey, and the mean value was 525.1 鹵72.35mg GAE / KG. The mean DPPH clearance rate (IC50) and ORAC of jujube nectar were 32.55 鹵4.16 mg/mL and 3.39 鹵0.85 渭 mol TEP 路g, which were significantly higher than that of litchi honey, and had a positive correlation with total phenol content. Therefore, the effect of jujube nectar on apoptosis of HepG2 human hepatoma cells was studied, and the mechanism of inducing apoptosis was explored. The results showed that the cell viability of HepG2 treated with jujube nectar decreased with the increase of jujube nectar concentration. Jujube nectar could induce the apoptosis of HepG2 cells, and the apoptosis characteristics were obvious. The proportion of apoptotic cells was positively correlated with the concentration of jujube nectar. In addition, jujube nectar could decrease mitochondrial membrane potential, increase the expression of Bax and Bad, and decrease the expression of Bcl-2 and Bcl-xL. At the same time, the expression of tumor suppressor gene p53 was increased with the increase of treatment concentration, and the expression of caspase-9 and downstream protein caspase-3 was up-regulated, which resulted in apoptosis. Therefore, jujube nectar may induce apoptosis of HepG2 cells through mitochondrial dependent pathway mediated by p53.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S896.1;TS201.2

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