中国新药杂志

山豆根多糖的制备、单糖组成及体外抗肿瘤作用

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王淑娜 , 杨丽雅 , 余登香 , 傅月朦 , 邓中平

中国新药杂志 | 实验研究 2023,32(11): 1155-1162

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中国新药杂志 | 实验研究 2023, 32(11): 1155-1162

山豆根多糖的制备、单糖组成及体外抗肿瘤作用

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王淑娜, 杨丽雅, 余登香, 傅月朦, 邓中平

作者信息

上海中医药大学创新中药研究院，上海 201203

王淑娜，女，硕士研究生，主要从事中药抗肿瘤作用研究。E-mail: wangshunalala@163.com。

通讯作者:

邓中平，男，博士生导师，研究员，主要从事中药药理学与毒理学研究。E-mail: dengzpcn@163.com。

Preparation, monosaccharide composition and in vitro antitumor effect of polysaccharide from Sophora tonkinensis Radix et Rhizoma

Shu-na WANG, Li-ya YANG, Deng-xiang YU, Yue-meng FU, Zhong-ping DENG

Affiliations

Innovation Research Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

出版时间: 2023-06-15

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摘要

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目的：

制备山豆根多糖（polysaccharide of Sophorae Tonkinensis Radix et Rhizoma, SRP）并分析其单糖组成，探索SRP的体外抗肿瘤作用。

方法：

水提醇沉法制备SRP，紫外-可见分光光度法测定SRP中多糖、黄酮、皂苷、酚类和蛋白质含量，高效液相色谱法测定生物碱含量，返滴定法测定有机酸含量。通过高效液相色谱仪，色谱柱采用Waters X Bridge C₁₈柱（250 mm×4.6 mm，5 μm），以乙腈：磷酸盐缓冲液（0.05 mol·L^-1，pH=6.8）=18∶82为流动相，等梯度洗脱，流速为0.8 mL·min^-1，柱温为30 ℃，检测波长为245 nm，建立SRP的单糖组成分析方法，测定SRP中各单糖含量并计算摩尔比。采用MTT法检测SRP抑制肿瘤细胞增殖的活性。

结果：

水提醇沉法制得SRP，棕黄色粉末，得率为3.95%，其中不含生物碱成分，多糖、黄酮、皂苷、有机酸和酚类含量分别为66.08%，7.17%，3.45%，2.54%和4.33%，蛋白质含量可忽略不计。SRP由甘露糖、鼠李糖、葡糖醛酸、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖组成，各单糖的摩尔比为1.25∶0.31∶0.28∶0.50∶40.47∶1.00∶10.46，SRP的质量分数为64.97%。SRP对Hepa1-6，HepG2，HT29和4T1细胞均具有显著的增殖抑制作用且呈浓度依赖性，IC₅₀值分别为770，1 910，1 870和1 880 μg·mL^-1。

结论：

本研究制备的SRP中多糖含量大于60%，已知成分含量大于80%，其中不含生物碱。1-苯基-3-甲基-5-吡唑啉酮（1-phenyl-3-methyl-5-pyrazolo, PMP）柱前衍生化-HPLC法精密度、重复性和准确性良好，可用于分析SRP的单糖组成。SRP是一类以葡萄糖和阿拉伯糖为主要成分的杂多糖。SRP具有明显的细胞毒作用，提示SRP可通过抑制肿瘤细胞增殖发挥抗肿瘤作用。本研究为推进SRP成分的基础研究和促进SRP的抗肿瘤作用研究提供了参考。

关键词

山豆根多糖 / 单糖组成 / Hepa1-6细胞 / HepG2细胞 / HT29细胞 / 4T1细胞 / 抗肿瘤

Abstract

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Objective:

To extract and separate polysaccharide from Sophora tonkinensis Radix et Rhizoma (SRP), determine the monosaccharide composition and content of SRP, and investigate the antitumor effects of SRP in vitro.

Methods:

SRP was prepared by water extraction and alcohol precipitation. The contents of polysaccharides, flavonoids, saponins, total phenols and proteins in SRP were determined by UV-Vis spectrophotometry, the contents of alkaloids were determined by high performance liquid chromatography (HPLC), and the content of organic acid was measured by back titration method. The monosaccharide analysis method of SRP was established by HPLC. A Waters X Bridge C₁₈ column (250 mm×4.6 mm,5 μm) was used with acetonitrile: phosphate buffer (0.05 mol·L^-1, pH=6.8) at the ration of 18∶82 as mobile phase, the flow rate was 0.8 mL·min^-1, the column temperature was 30 ℃, and the detection wavelength was 245 nm. The contents of monosaccharides in SRP were determined and the mole ratio was calculated. MTT assay was used to detect the antitumor effects of SRP in vitro.

Results:

SRP, which is brown powder, was obtained by water extraction and alcohol precipitation with a yield of 3.95%. The contents of polysaccharides, flavonoids, saponins, organic acids and total phenols in SRP were 66.08%, 7.17%, 3.45%, 2.54% and 4.33%, respectively. No alkaloids and proteins were detected. SRP was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose. The mole ratios were 1.25∶0.31∶0.28∶0.50∶40.47∶1.00∶10.46, and the percentage of polysaccharide was 64.97%. SRP had inhibitory effect on Hepa1-6, HepG2, HT29 and 4T1 cells in a concentration-dependent manner with IC₅₀ of 770, 1 910, 1 870 and 1 880 μg·mL^-1, respectively.

Conclusion:

The SRP prepared in this study contained more than 60% polysaccharide and more than 80% known components, among which no alkaloids were detected. HPLC with pre-column derivatization of 1-phenyl-3-methyl-5-pyrazolo has good precision, reproducibility and accuracy, and is suitable for the monosaccharide composion analysis of SRP polysaccharides. SRP is a kind of heteroglycan with glucose and arabinose as the main components. SRP has significant cytotoxic effect by inhibiting tumor cell proliferation, suggesting that it may play a anti-tumor role. This study provides a reference for advancing the basic research of the composition and antitumor effect of SRP.

Key words

Sophorae Tonkinensis Radix et Rhizoma polysaccharide / monosaccharide composition / Hepa1-6 cells / HepG2 cells / HT29 cells / 4T1 cells / antitumor

引用本文

王淑娜, 杨丽雅, 余登香, 傅月朦, 邓中平. 山豆根多糖的制备、单糖组成及体外抗肿瘤作用. 中国新药杂志, 2023 , 32 (11) : 1155 -1162 .

Shu-na WANG, Li-ya YANG, Deng-xiang YU, Yue-meng FU, Zhong-ping DENG. Preparation, monosaccharide composition and in vitro antitumor effect of polysaccharide from Sophora tonkinensis Radix et Rhizoma[J]. Chinese Journal of New Drugs, 2023 , 32 (11) : 1155 -1162 .

参考文献

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2023年第32卷第11期

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首发时间：2026-03-06
出版时间：2023-06-15

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录用日期：2022-07-13

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作者信息

上海中医药大学创新中药研究院，上海 201203

通讯作者:

邓中平，男，博士生导师，研究员，主要从事中药药理学与毒理学研究。E-mail: dengzpcn@163.com。

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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