牡丹籽粕多糖理化特性及抗氧化、抗腫瘤功能研究
發(fā)布時(shí)間：2018-10-08 22:06
【摘要】：牡丹除了具有觀賞價(jià)值外,常被用作藥材,具有抗氧化、免疫調(diào)節(jié)等多種功能特性。采用連續(xù)提取的方法制備了牡丹籽粕4種多糖,測(cè)定了4種多糖的理化性質(zhì),研究了流變學(xué)特性和乳化活性,分析了抗氧化活性和抗腫瘤活性,并對(duì)牡丹籽粕多糖4種組分的上述性質(zhì)進(jìn)行了比較。研究結(jié)果為進(jìn)一步把牡丹籽粕多糖開(kāi)發(fā)成功能食品提供了理論基礎(chǔ)。牡丹籽粕經(jīng)熱緩沖液、螯合劑、稀堿溶液和濃堿溶液連續(xù)提取后得到4種多糖組分:HBSS、CHSS、DASS和CASS。傅里葉紅外光譜分析表明牡丹籽粕多糖在3600 3200 cm~(-1)和1200 800 cm~(-1)呈現(xiàn)糖類(lèi)的特征吸收峰。高效液相色譜法測(cè)定提取的4種多糖組分的分子量分別為3467、4677、229和56 kDa。HBSS主要由甘露糖和葡萄糖組成,CHSS主要由半乳糖和阿拉伯糖組成,DASS主要由阿拉伯糖、半乳糖和甘露糖組成,CASS主要由半乳糖、阿拉伯糖和葡萄糖組成。4種牡丹籽粕多糖的熱學(xué)特性稍有差別,CASS顯示出了最高的峰值溫度和熱焓。研究了牡丹籽粕多糖的靜態(tài)流變性質(zhì)和動(dòng)態(tài)流變性質(zhì)�？疾炝藵舛取H、溫度、鹽離子種類(lèi)和濃度對(duì)牡丹籽粕多糖水溶液流變學(xué)特性的影響。4種牡丹籽粕多糖水溶液表現(xiàn)為典型的非牛頓流體,具有剪切稀化的特征,HBSS的黏度最高。當(dāng)pH變?yōu)?.0或10.0時(shí),CASS的表觀黏度下降最明顯;加入Ca~(2+)后牡丹籽粕多糖水溶液略有降低;在90°C的高溫下,多糖水溶液表觀黏度略有下降,CASS降幅最小;而Na+的加入使得CHSS黏度明顯上升,-20°C冷凍處理會(huì)使多糖溶液的表觀黏度增加。隨剪切振蕩頻率(0.01-100 Hz)的增大,不同濃度的多糖溶液的儲(chǔ)能模量G’和損耗模量G”都表現(xiàn)出連續(xù)升高。隨著牡丹籽粕多糖溶液濃度的增加,4種組分的多糖溶液交叉頻率有所降低。牡丹籽粕多糖對(duì)油水溶液的乳化活性與乳化穩(wěn)定性具有濃度依賴性,HBSS乳化活性最強(qiáng),CHSS乳化穩(wěn)定時(shí)間最長(zhǎng)。4種牡丹籽粕多糖組分在抗氧化活性上有很大的差異,CASS能有效地清除DPPH和ABTS自由基,還原Fe~(3+)離子,HBSS對(duì)羥基自由基的清除能力最強(qiáng),CHSS對(duì)亞鐵離子的螯合能力最強(qiáng)。牡丹籽粕多糖對(duì)HCT-116、PC-3和Hela細(xì)胞的細(xì)胞毒性、細(xì)胞周期、細(xì)胞凋亡的影響被研究。CASS具有最強(qiáng)的抗腫瘤活性,與對(duì)照組5-FU相比,CASS顯示出了對(duì)Hela細(xì)胞更強(qiáng)的毒性作用,進(jìn)一步研究表明,CASS能夠使HCT-116、PC-3和Hela細(xì)胞的生長(zhǎng)周期阻滯于G0/G1期,除此以外,用CASS處理過(guò)的Hela細(xì)胞,細(xì)胞凋亡率達(dá)到30.94%。以上結(jié)果表明,CASS具有潛在的抗氧化和抗腫瘤活性,將來(lái)可能在功能食品中應(yīng)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 擄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 擄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.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：TS229
本文編號(hào)：2258427
[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 擄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 擄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.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：TS229
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