黑木耳多糖模拟水解物的抗氧化及胃肠道分布规律研究
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摘要
探讨体外人工胃液肠水解黑木耳多糖片段的抗氧化活性及其在小鼠消化道内的动态分布规律。依次采用碱提醇沉,脱蛋白,人工胃肠液水解,透析,凝胶柱层析(Sephadex G100)得到5种黑木耳多糖组分(AAPF-1,AAPF-2,AAPF-3,AAPF-4,AAPF-5),利用体外OH·和DPPH·清除率和线虫H2O2抗性实验分析5种多糖的活性。利用异硫氰酸荧光素标记黑木耳多糖获得AAPF-1-Tyr-FITC,追踪其在小鼠胃肠道内24 h的动态分布规律。结果表明,5种黑木耳多糖组分中,AAPF-1(0.4 mg/m L)对OH·与DPPH·清除率均为最高,分别达68.54%与42.18%,并在60~150 min内均可显著延长线虫寿命。AAPF-1经胃很快排空到小肠,在小鼠小肠基本不吸收,而在大肠中的存留时间在24 h以上。
The purpose of this study was to investigate the antioxidant activity and the dynamic distribution of the digestive tract in mice of the Auricularia auricular polysaccharide fragments from the artificial gastrointestinal fluid in vitro. The 5 kinds of polysaccharides(AAPF-1,AAPF-2,AAPF-3,AAPF-4,AAPF-5) were separated from Auricularia auricular by alkali extraction and alcohol precipitation, deproteinization, artificial gastrointestinal fluid hydrolysis, dialysis and gel column chromatography(Sephadex G100). OH· and DPPH· clearance in vitro and nematode H2 O2 resistance experiment of those 5 kinds of polysaccharides were analyzed. AAPF-1-Tyr-FITC was obtained by fluorescein labeled, and the dynamic distribution in the gastrointestinal tract of mice was studied. Results showed that AAPF-1(0.4 mg/m L) for OH· and DPPH· removal rate was the highest among 5 kinds of polysaccharides, that was 68.54% and 42.18% respectively, and the life span of the nematodes was significantly prolonged within 60~150 min. AAPF-1 was quickly evacuated to the small intestine, and not absorbed in the small intestine in mice, and the remaining time in the large intestine was more than 24 h.
The purpose of this study was to investigate the antioxidant activity and the dynamic distribution of the digestive tract in mice of the Auricularia auricular polysaccharide fragments from the artificial gastrointestinal fluid in vitro. The 5 kinds of polysaccharides(AAPF-1,AAPF-2,AAPF-3,AAPF-4,AAPF-5) were separated from Auricularia auricular by alkali extraction and alcohol precipitation, deproteinization, artificial gastrointestinal fluid hydrolysis, dialysis and gel column chromatography(Sephadex G100). OH· and DPPH· clearance in vitro and nematode H2 O2 resistance experiment of those 5 kinds of polysaccharides were analyzed. AAPF-1-Tyr-FITC was obtained by fluorescein labeled, and the dynamic distribution in the gastrointestinal tract of mice was studied. Results showed that AAPF-1(0.4 mg/m L) for OH· and DPPH· removal rate was the highest among 5 kinds of polysaccharides, that was 68.54% and 42.18% respectively, and the life span of the nematodes was significantly prolonged within 60~150 min. AAPF-1 was quickly evacuated to the small intestine, and not absorbed in the small intestine in mice, and the remaining time in the large intestine was more than 24 h.
引文
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[15]申建和,陈琼华.黑木耳多糖的抗血栓作用[J].中国药科大学学报,1990,21(1):39-42.
[16]樊一桥,武谦虎,盛健惠.黑木耳多糖抗血栓作用的研究[J].中国生化药物杂志,2009,30(6):410-412.
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[18]朱磊,王振宇.黑木耳多糖对小鼠抗疲劳作用的研究[J].营养学报,2008,30(4):430-432.
[19]张会新,刘洪雨,刘畅,等.黑木耳多糖对小鼠免疫功能的影响[J].动物医学进展,2009,30(7):23-25.
[20]张倩,周秋丽,胡剑江,等.荧光标记及检测技术在中药大分子活性示踪中的应用[J].中国实验方剂学杂志,2010,16(14):215-220.
[21]ARNOSTI C.Fluorescent derivatization of polysaccharides and carbohydrate-containing biopolymers for measurement of enzyme activities in complex media[J].Journal of Chromatography B,2003,793(1):181-191.
[22]ZHANG Y,WANG Z X,ZHANG X R,et al.One-pot fluorescent labeling of saccharides with fluorescein-5-thiosemicarbazide for imaging polysaccharides transported in living cells[J].Carbohydrate Research,2011,346(14):2156-2164.
[2]MA Z C,WANG J G,ZHANG L,et al.Evaluation of water solubleβ-D-glucan from Auricularia auricular-judae as potential anti-tumor agent[J].Carbohydrate Polymers,2010,80(3):977-983.
[3]MISAKI A,KAKUTA M,SASAKI T,et al.Studies on interrelation of structure and antitumor effects of polysaccharides:antitumor action of periodate modified,branched(1→3)-β-D-glucan of Auricularia Auricula-judae,and polysaccharides confaining(1→3)glycosidic linkages[J].Carbohydrate Research,1981,92(1):115-120.
[4]周学君,俞发.毛木耳多糖的抗氧化作用[J].中国医院药学杂志,2000,20(10):610-611.
[5]史亚丽,姜川,杨立红,等.黑木耳粗多糖对力竭小鼠抗氧化能力的影响[J].现代预防医学,2008,35(24):4845-4847.
[6]申进文,冯雅岚,余海尤,等.硫酸化分级香菇多糖抗氧化活性研究[J].菌物学报,2010,29(3):449-453.
[7]李兴泰,高明波.黑木耳多糖清除活性氧及保护线粒体[J].食品科学,2004,25(1):171-173.
[8]田志杰,吕世杰,姜艳霞,等.正交法提取木耳多糖及对大鼠血清超氧化物歧化酶和体外肝脏脂质过氧化的影响[J].时珍国医国药,2009,20(11):2694-2695.
[9]韩春然,马永强,唐娟.黑木耳多糖的提取及降血糖作用[J].食品与生物技术学报,2006,25(5):111-114.
[10]CHEUNG P C K.The hypocholesterolemic effect of two edible mushrooms:Auricularia auricular(tree-ear)and Tremella fuciformis(white jelly-leaf)in hypercholesterolemic rats[J].Nutrition Research,1996,16(10):1721-1725.
[11]周国华,于国萍.黑木耳多糖降血脂作用的研究[J].现代食品科技,2005,21(1):46-48.
[12]蔡小玲,章佩芬,何有明,等.黑木耳多糖、红菇多糖的降胆固醇作用研究[J].深圳中西医结合杂志,2002,12(3):137-139.
[13]谢意珍,周静文,李崇.黑木耳液体发酵条件的优化及发酵物的降血脂功能[J].食用菌学报,2008,15(2):47-50.
[14]杨春瑜,姜启兴,夏文水,等.黑木耳超微粉多糖相对分子质量分布及降血脂功能研究[J].中国食品学报,2008,8(6):23-32.
[15]申建和,陈琼华.黑木耳多糖的抗血栓作用[J].中国药科大学学报,1990,21(1):39-42.
[16]樊一桥,武谦虎,盛健惠.黑木耳多糖抗血栓作用的研究[J].中国生化药物杂志,2009,30(6):410-412.
[17]樊黎生,龚晨睿,张声华.黑木耳多糖抗辐射效应的动物实验[J].营养学报,2005,27(6):525-526.
[18]朱磊,王振宇.黑木耳多糖对小鼠抗疲劳作用的研究[J].营养学报,2008,30(4):430-432.
[19]张会新,刘洪雨,刘畅,等.黑木耳多糖对小鼠免疫功能的影响[J].动物医学进展,2009,30(7):23-25.
[20]张倩,周秋丽,胡剑江,等.荧光标记及检测技术在中药大分子活性示踪中的应用[J].中国实验方剂学杂志,2010,16(14):215-220.
[21]ARNOSTI C.Fluorescent derivatization of polysaccharides and carbohydrate-containing biopolymers for measurement of enzyme activities in complex media[J].Journal of Chromatography B,2003,793(1):181-191.
[22]ZHANG Y,WANG Z X,ZHANG X R,et al.One-pot fluorescent labeling of saccharides with fluorescein-5-thiosemicarbazide for imaging polysaccharides transported in living cells[J].Carbohydrate Research,2011,346(14):2156-2164.