白及多糖BSP-1的分离纯化、结构表征及抗肿瘤活性研究
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摘要
目的从白及Bletillastriata块茎中分离纯化多糖BSP-1,并对其结构和抗肿瘤活性进行研究。方法将提取的粗多糖经DEAE-cellulose柱色谱分离得到3个多糖组分BP-1、BP-2、BP-3。BP-1通过SephadexG-200柱色谱纯化后得到组分BSP-1。采用HPGPC、IR、GC、GC-MS、甲基化及~1H-、~(13)C-NMR等方法对该多糖的结构进行表征,并考察BSP-1抗肿瘤活性。结果测得BSP-1相对分子质量为4.72×10~5,由葡萄糖和甘露糖组成。主链由β-1,4甘露糖,侧链由末端连接的葡萄糖组成,其物质的量比为8∶1。体外肿瘤细胞活性实验表明,BSP-1能抑制肝癌Hep G2细胞的增殖。BSP-1体内可显著抑制裸鼠的瘤质量,抑制率为66.42%。结论 BSP-1的结构确定为进一步阐明和开发白及中多糖类成分提供了一定的参考。
Objective Bletilla striata polysaccharide(BSP-1) extracted from the stem tubers of B. striata was purified to study the structural properties and antitumor activity. Methods The crude polysaccharide was fractionated by DEAE-cellulose column chromatography, and three fractions were obtained including BP-1, BP-2, and BP-3, respectively, BP-1 was further purified by SephadexG-200 column chromatography, and a sub-fraction named BSP-1 was obtained. Subsequently, HPGPC, IR spectroscopy, gas chromatography(GC), GC coupled to mass spectrometry(GC-MS) and methylation analysis, and ~1 H-and ~(13) C-NMR were employed to characterize the structural properties of the polysaccharide fractions. Moreover, the anticancer activity was investigated in vivo. Results The results showed that the molecular weight of BSP-1 were 4.72 × 10~5. BSP-1 was consisted of mannose(Man) and glucose(Glc) residues. The backbone of BSP-1 was composed of β-1,4-linked Man and Glc residues at the end with a molar ratio of 8∶1. BSP-1 could inhibit the tumor proliferation of HepG2-bearing mice. The tumor weight of mice was significantly inhibited by BSP-1 with inhibitory rate of 66.42%. Conclusion Structural characterization of BSP-1 provides the significant reference for the further development and elucidation of polysaccharides from B. striata as new drugs.
Objective Bletilla striata polysaccharide(BSP-1) extracted from the stem tubers of B. striata was purified to study the structural properties and antitumor activity. Methods The crude polysaccharide was fractionated by DEAE-cellulose column chromatography, and three fractions were obtained including BP-1, BP-2, and BP-3, respectively, BP-1 was further purified by SephadexG-200 column chromatography, and a sub-fraction named BSP-1 was obtained. Subsequently, HPGPC, IR spectroscopy, gas chromatography(GC), GC coupled to mass spectrometry(GC-MS) and methylation analysis, and ~1 H-and ~(13) C-NMR were employed to characterize the structural properties of the polysaccharide fractions. Moreover, the anticancer activity was investigated in vivo. Results The results showed that the molecular weight of BSP-1 were 4.72 × 10~5. BSP-1 was consisted of mannose(Man) and glucose(Glc) residues. The backbone of BSP-1 was composed of β-1,4-linked Man and Glc residues at the end with a molar ratio of 8∶1. BSP-1 could inhibit the tumor proliferation of HepG2-bearing mice. The tumor weight of mice was significantly inhibited by BSP-1 with inhibitory rate of 66.42%. Conclusion Structural characterization of BSP-1 provides the significant reference for the further development and elucidation of polysaccharides from B. striata as new drugs.
引文
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[18]陈冠,赵振宇,傅予,等.苦豆子多糖的结构表征及其对CT26抗肿瘤活性研究[J].中草药,2017,48(6):1103-1107.
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[22]Pang S Q,Wang G Q,Wu S,et al.Effects of polysaccharides from Bletillae Rhizoma on the protection in rats during deep second-degree burn wound healing[J].J Chin Med Mater,2013,36(11):1819-1823.
[2]赵永伟,王艳宏.白及多糖的研究进展[J].中国中医药科技,2015,22(4):479-481.
[3]吕洪乐,张同华,李倩.白及多糖药理作用的研究进展[J].中国药房,2015,26(28):4014-4016.
[4]李春雪,曾锐,钱妍,等.多孔性白及胶的制备及辅料特性初步研究[J].中草药,2017,48(20):4245-4252.
[5]雷震,常明泉,陈黎,等.白及的临床应用研究进展[J].中国药师,2013,16(8):1240-1242.
[6]Qian J,Vossoughi D,Woitaschek D,et al.Combined transarterial chemoembolization and arterial administration of Bletilla striata in treatment of liver tumor in rats[J].World J Gastroenterol,2003,9(12):2676-2680.
[7]王程成,崔莉,张振海,等.口服中药多糖调节肿瘤免疫应答研究思路[J].中国中药杂志,2016,41(10):1965-1971.
[8]李珊珊,金银萍,姚春林,等.人参多糖的结构与活性研究进展[J].中国中药杂志,2014,39(24):4709-4715.
[9]张蔷,高文远,满淑丽.黄芪中有效成分药理活性的研究进展[J].中国中药杂志,2012,37(2):3203-3207.
[10]Feeeeira I C,Heleno S A,Reis F S,et al.Chemical features of Ganoderma polysaaccharides with antioxidant,antitumor and antimicrobial activitives[J].Phytochemistry,2015,114:38-55.
[11]芮海云,吴国荣,陈景耀,等.白芨中性杂多糖的分离纯化与结构解析[J].安徽农业大学学报,2004,31(1):30-33.
[12]王博,徐莎,黄琳娟,等.白芨多糖BSPI-A的分离纯化及结构研究[J].食品科学,2010,31(37):120-123.
[13]魏绍云,齐惠玲,王继轮,等.苯酚-硫酸法测定白及多糖[J].天津化工,2000,14(3):35-36.
[14]俞晓玉,王树威,白晓燕,等.粮食中蛋白质含量的测定[J].广东化工,2017,44(4):114-115.
[15]韩婷,史国华,李素哲,等.壳聚糖中蛋白质含量测定方法的研究[J].中国医疗器械杂志,2016,40(2):122-124.
[16]Lehrfeld J.Simultaneous gas-liquid chromatographic determination of aldonic acids and aldoses[J].Anal Chem,1985,57:346-347.
[17]张秀娟,杨姗姗.半枝莲多糖体内抗肿瘤及其免疫调节作用的实验研究[J].亚太传统医药,2008,4(2):54-56.
[18]陈冠,赵振宇,傅予,等.苦豆子多糖的结构表征及其对CT26抗肿瘤活性研究[J].中草药,2017,48(6):1103-1107.
[19]Zhang X,Yu L,Bi H,et al.Total fractionation and characterization of the water-soluble polysaccharides isolated from Panax ginseng C.A.Meyer[J].Carbohydrate Polymers,2009,77(3):544-552.
[20]Park J E,Woo K W,Choi S U,et al.Two new cytotoxic spirostane-steroidal saponins from the roots of Bletilla striata[J].Helv Chim Acta,2014,97(1):56-63.
[21]谭西,周欣,陈国华.植物多糖构效关系研究进展[J].中国药学杂志,2017,42(21):4104-4109.
[22]Pang S Q,Wang G Q,Wu S,et al.Effects of polysaccharides from Bletillae Rhizoma on the protection in rats during deep second-degree burn wound healing[J].J Chin Med Mater,2013,36(11):1819-1823.
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