牡丹籽粕多糖理化特性及抗氧化、抗肿瘤功能研究

发布时间：2018-10-08 22:06

【摘要】：牡丹除了具有观赏价值外,常被用作药材,具有抗氧化、免疫调节等多种功能特性。采用连续提取的方法制备了牡丹籽粕4种多糖,测定了4种多糖的理化性质,研究了流变学特性和乳化活性,分析了抗氧化活性和抗肿瘤活性,并对牡丹籽粕多糖4种组分的上述性质进行了比较。研究结果为进一步把牡丹籽粕多糖开发成功能食品提供了理论基础。牡丹籽粕经热缓冲液、螯合剂、稀碱溶液和浓碱溶液连续提取后得到4种多糖组分:HBSS、CHSS、DASS和CASS。傅里叶红外光谱分析表明牡丹籽粕多糖在3600 3200 cm~(-1)和1200 800 cm~(-1)呈现糖类的特征吸收峰。高效液相色谱法测定提取的4种多糖组分的分子量分别为3467、4677、229和56 kDa。HBSS主要由甘露糖和葡萄糖组成,CHSS主要由半乳糖和阿拉伯糖组成,DASS主要由阿拉伯糖、半乳糖和甘露糖组成,CASS主要由半乳糖、阿拉伯糖和葡萄糖组成。4种牡丹籽粕多糖的热学特性稍有差别,CASS显示出了最高的峰值温度和热焓。研究了牡丹籽粕多糖的静态流变性质和动态流变性质。考察了浓度、pH、温度、盐离子种类和浓度对牡丹籽粕多糖水溶液流变学特性的影响。4种牡丹籽粕多糖水溶液表现为典型的非牛顿流体,具有剪切稀化的特征,HBSS的黏度最高。当pH变为4.0或10.0时,CASS的表观黏度下降最明显;加入Ca~(2+)后牡丹籽粕多糖水溶液略有降低;在90°C的高温下,多糖水溶液表观黏度略有下降,CASS降幅最小;而Na+的加入使得CHSS黏度明显上升,-20°C冷冻处理会使多糖溶液的表观黏度增加。随剪切振荡频率(0.01-100 Hz)的增大,不同浓度的多糖溶液的储能模量G’和损耗模量G”都表现出连续升高。随着牡丹籽粕多糖溶液浓度的增加,4种组分的多糖溶液交叉频率有所降低。牡丹籽粕多糖对油水溶液的乳化活性与乳化稳定性具有浓度依赖性,HBSS乳化活性最强,CHSS乳化稳定时间最长。4种牡丹籽粕多糖组分在抗氧化活性上有很大的差异,CASS能有效地清除DPPH和ABTS自由基,还原Fe~(3+)离子,HBSS对羟基自由基的清除能力最强,CHSS对亚铁离子的螯合能力最强。牡丹籽粕多糖对HCT-116、PC-3和Hela细胞的细胞毒性、细胞周期、细胞凋亡的影响被研究。CASS具有最强的抗肿瘤活性,与对照组5-FU相比,CASS显示出了对Hela细胞更强的毒性作用,进一步研究表明,CASS能够使HCT-116、PC-3和Hela细胞的生长周期阻滞于G0/G1期,除此以外,用CASS处理过的Hela细胞,细胞凋亡率达到30.94%。以上结果表明,CASS具有潜在的抗氧化和抗肿瘤活性,将来可能在功能食品中应用。
[Abstract]:In addition to ornamental value, peony is often used as medicinal materials, with antioxidant, immune regulation and other functional characteristics. Four polysaccharides from peony seed meal were prepared by continuous extraction. The physicochemical properties of the four polysaccharides were determined, the rheological properties and emulsifying activities were studied, and the antioxidant and anti-tumor activities were analyzed. The above properties of four polysaccharides from peony seed meal were compared. The results provide a theoretical basis for the further development of peony seed meal polysaccharides into functional foods. After continuous extraction of peony seed meal from hot buffer solution, chelating agent, dilute alkali solution and concentrated alkali solution, four polysaccharide fractions were obtained, I. e., 1: HBSS-CHSS-DASS and CASS.. Fourier transform infrared spectroscopy (FTIR) analysis showed that the polysaccharides of peony seed meal showed the characteristic absorption peaks of carbohydrate at 3600 ~ 3200 cm~ (-1) and 1200 ~ 800 cm~ (-1). The molecular weights of the four polysaccharides extracted by HPLC were 346747 7229 and 56 kDa.HBSS, respectively, which were mainly composed of mannose and glucose, mainly galactose and arabinose. Cass consisted mainly of galactose, arabinose and glucose. The thermal properties of polysaccharides from 4 species of peony seed meal were slightly different. Cass showed the highest peak temperature and enthalpy. The static and dynamic rheological properties of polysaccharides from peony seed meal were studied. The effects of pH, temperature, salt ion species and concentration on rheological properties of polysaccharide aqueous solution of peony seed meal were investigated. 4 species of polysaccharide aqueous solution of peony seed meal showed typical non-Newtonian fluid, and the viscosity of HBSS was the highest with the characteristic of shear-thinning. When pH changed to 4.0 or 10.0, the apparent viscosity of Cass decreased most obviously, the aqueous solution of polysaccharides from peony seed meal decreased slightly after adding Ca~ (2), the apparent viscosity of Cass decreased slightly at 90 掳C and the decrease of Cass was the least. However, the addition of Na significantly increased the viscosity of CHSS solution after freezing at -20 掳C, which resulted in an increase in the apparent viscosity of polysaccharide solution. With the increase of shearing oscillation frequency (0.01-100 Hz), the storage modulus G 'and the loss modulus G "of different concentrations of polysaccharide solution increased continuously. With the increase of the concentration of polysaccharide solution of peony seed meal, the cross frequency of polysaccharide solution of four components decreased. Emulsifying activity and emulsifying Stability of polysaccharides from Peony seed meal in concentration dependent manner HBSS has the strongest emulsifying activity and the longest emulsifying and stabilizing time of CHSS. 4 kinds of Peony seed meal polysaccharides have great difference in antioxidant activity and Cass energy. Effectively scavenging DPPH and ABTS radicals, The scavenging ability of Fe~ (3) ion to hydroxyl radical is the strongest, and the chelating ability of Fe~ (3) ion to ferrous ion is the strongest. The effects of polysaccharides from peony seed meal on the cytotoxicity, cell cycle and apoptosis of HCT-116,PC-3 and Hela cells were studied. Cass had the strongest antitumor activity. Compared with the control group, Cass showed stronger cytotoxicity to Hela cells. Further studies showed that Cass could block the growth cycle of HCT-116,PC-3 and Hela cells in G0/G1 phase. In addition, the apoptosis rate of Hela cells treated with CASS could reach 30.944%. These results suggest that Cass has potential antioxidant and antitumor activities and may be used in functional foods in the future.
【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS229

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