黑果枸杞叶多糖LRLP3的结构、抗氧化活性及免疫活性

doi:10.7503/cjcu20150690

摘要/Abstract

摘要：

黑果枸杞叶经水提醇沉, 离子交换柱层析和凝胶柱层析分离纯化, 得到平均分子量为79400的均一多糖组分LRLP3. 对该多糖的理化性质、结构、抗氧化活性及免疫活性的研究结果表明, LRLP3为多分支结构, 主链为(1→3)βGalp, 大部分半乳糖6位存在分支; 支链由(1→6)βGalp, (1→4)βGalp, (1→3)βAraf, (1→3)αArap, (1→5)βAraf和(1→2,4)αRhap组成, 非还原末端由αAraf, βGalp和βGlcp组成. LRLP3具有较强的还原能力, 可显著清除1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟自由基和超氧阴离子自由基, 有效抑制Cu²⁺/H₂O₂诱导的蛋白氧化损伤和H₂O₂诱导的细胞氧化损伤. LRLP3在体外对未经诱导和经刀豆蛋白(ConA)或脂多糖(LPS)诱导的小鼠脾细胞增殖均有促进作用.

关键词: 黑果枸杞叶, 多糖, 结构分析, 抗氧化活性, 免疫活性

Abstract:

A combination of chemical and instrumental analysis was performed to investigate the structural characterization and biological activity of a polysaccharide LRLP3 which was isolated from the Lycium ruthenicum leaves. The results demonstrated that LRLP3 was a highly branched polysaccharide with a backbone of (1→3)-linked-βGalp substituted at C-6 position by galactosyl. The branches were composed of(1→6)-linked-βGalp,(1→4)-linked-βGalp,(1→3)-linked-αAraf and βArap,(1→5)-linked-αAraf, and (1→2, 4)-linked αRhap, and the terminal residues were αAraf, βGalp and βGlcp. Additional, LRLP3 had strong reducing power, could significantly scavenge DPPH, hydroxyl and superoxide free radical, could effectually inhibit Cu²⁺/H₂O₂ induced protein damage and H₂O₂ induced cell damage in vitro. Meanwhile, immunological assay showed that LRLP3 could stimulate proliferation of spleen lymphocytes significantly with or without mitogens(ConA or LPS) in vitro. Therefore, LRLP3 was a natural arabinogalactan which had the potential function of antioxidant and immunoloregulation.

Key words: Lyciumruthenicum Murray leaves, Polysaccharide, Structural analysis, Antioxidant activity, Immunological activity

中图分类号:

O629.12

TrendMD:

刘洋, 殷璐, 龚桂萍, 彭艺芳, 黄琳娟, 王仲孚. 黑果枸杞叶多糖LRLP3的结构、抗氧化活性及免疫活性. 高等学校化学学报, 2016, 37(2): 261.

LIU Yang, YIN Lu, GONG Guiping, PENG Yifang, HUANG Linjuan, WANG Zhongfu. Structural Characterization, Antioxidant Activity and Immunomodulatory Activity of the Polysaccharide LRLP3 from Leaves of Lycium ruthenicum Murra^†. Chem. J. Chinese Universities, 2016, 37(2): 261.

图/表 6

Table 1 Partially O-methylated alditol acetates of LRLP3 and LRLP3-I

Peak	Partially methylated sugar	Deduced linkage	Relative molar ratio
Peak	Partially methylated sugar	Deduced linkage	LRLP3	LRLP3-I
1	2,3,5-Me₃-Ara	Araf(1-	7
2	2,5-Me₂-Ara	-3)Araf(1-	4
3	2,3-Me₂-Ara	-5)Araf(1-	12
4	2,3,4,6-Me₄-Glc	Glcp(1-	1
5	2,3,4,6-Me₄-Gal	Galp(1-	2	5
6	2,4-Me₂-Ara	-3)Arap(1-	11
7	3-Me-Rha	-2,4)Rhap(1-	2
8	2,3,6-Me₃-Gal	-4)Galp(1-	3
9	2,4,6-Me₃-Gal	-3)Galp(1-	3	3
10	2,3,4-Me₃-Gal	-6)Galp(1-	1	4
11	2,4-Me₂-Gal	-3,6)Galp(1-	8	5

Fig.1 ESI-MS analysis of LRLP3-O

Fig.2 Hypothetical structure of the repeat unit of LRLP3

Fig.3 Antioxidant effect in vitro of LRLP3 (A) Reducing power of LRLP3; (B) scavenging effect on DPPH radical by LRLP3; (C) scavenging effect on hydroxyl radical by LRLP3; (D) scavenging effect on superoxide radical by LRLP3. The results were from three independent experiments and expressed as means±S.D.

Fig.4 Protective effect of LRLP3 on Cu2+/H2O2-induced BSA protein damage(A) and H2O2-induced HeLa cells damage(B) The results were obtained from three independent experiments and expressed as means ± S.D. **P<0.01 compared with control group; *** P<0.001 compared with control group; “-”means “BSA protein or HeLa cells were not treated with H2O2 or Cu2+/H2O2 or LRLP3”; “+” means “BSA protein or HeLa cells were treated with H2O2 or Cu2+/H2O2 or LRLP3”.

Fig.5 In vitro effects of LRLP3 on splenocyte proliferation index(A), ConA-induced splenocyte proliferation(B) and LPS-induced splenocyte proliferation(C) The results were from three independent experiments and expressed as means ± S.D. **P<0.01 compared with control group; “-”means “splenocytes were not treated with LPS or ConA or LRLP3”; “+” means “splenocytes were treated with LPS or ConA or LRLP3”.

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