Structural characterization, antioxidant and antitumor properties of an acidic polysaccharide from toad venom

IF 5.2 2区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Chemical and Biological Technologies in Agriculture Pub Date : 2025-09-01 DOI:10.1186/s40538-025-00842-7

Feng Pan, Xinting Zhu, Nan Yang, Kunmei Li, Gejie Shi, Qinghong Kong, Sanhua Li, Ying Qing, Linjie Meng, Yun Liu

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引用次数: 0

Abstract

Background

As a valuable agricultural product derived primarily from cultivated toads, toad venom has been widely used for its significant antitumor properties in pharmacological formulations. However, the structural and biological characteristics of its major constituents, polysaccharides, remain poorly understood, limiting the development of innovative antitumor drugs.

Results

In this study, an extracellular acidic homogeneous polysaccharide (TvPs3-2, average molecular mass: 3.43 × 10² kDa) is isolated from toad venom. Compositional analysis by 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivatization high-performance liquid chromatography (PMP-HPLC) reveals that TvPs3-2 consists of mannose (Man), glucuronic acid (GlcA), glucose (Glc), galactose (Gal), arabinose (Ara), and fucose (Fuc) in a molar ratio of 9.30:26.52:17.26:2.20:8.40:36.31. Comprehensive structural characterization through methylation analysis combined with extensive nuclear magnetic resonance (NMR) spectroscopy indicates a possible backbone: → 4)-β-D-GlcAp-(1 → 3,6)-β-D-Galp-(1 → 3)-α-L-Fucp-(1 → 4)-α-D-Manp-(1 → 2)-β-D-Glcp-(1 → 2,4)-α-D-Glcp-(1 → 4)-β-D-Glcp-(1 → 2)-α-L-Fucp-(1 → 4)-β-D-GlcAp-(1 → , with multiple branched segments. Morphologically, TvPs3-2 exhibits interconnected bulges, undulating surfaces, and square/rhombic crystals while maintaining a triple-helix conformation. Additionally, TvPs3-2 exhibits good antioxidant activity, particularly against 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS, half maximal inhibitory concentration (IC₅₀) = 0.36 mg/mL), alongside moderate 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging effects. Furthermore, TvPs3-2 demonstrates notable antiproliferative effects on lung (A549), liver (SK-Hep1), and colon (HCT116 and HT-29) cancer cell lines, with the highest efficacy against SK-Hep1 hepatoma cells (IC₅₀ = 1.6 mg/mL), mediated by apoptosis induction, migration inhibition, and modulation of metabolic/oxidative stress pathways, while it demonstrates relatively low inhibitory activity against healthy H9C2(2-1) cells.

Conclusions

This study comprehensively characterizes a novel acidic polysaccharide (TvPs3-2) from toad venom with remarkable antioxidant and antitumor activities, providing a molecular basis for its future application in anticancer drug development.

Graphical Abstract

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蟾蜍毒液酸性多糖的结构表征、抗氧化和抗肿瘤特性

蟾蜍毒是一种主要从栽培蟾蜍中提取的有价值的农产品，因其显著的抗肿瘤特性而被广泛应用于药理制剂中。然而，其主要成分多糖的结构和生物学特性仍然知之甚少，限制了创新抗肿瘤药物的开发。结果从蟾蜍毒液中分离得到胞外酸性均质多糖tvps2 -2，平均分子质量为3.43 × 102 kDa。通过柱前衍生化高效液相色谱（PMP- hplc）分析，tvps2 -2由甘露糖（Man）、葡萄糖醛酸（GlcA）、葡萄糖（Glc）、半乳糖（Gal）、阿拉伯糖（Ara）和焦糖（Fuc）组成，摩尔比为9.30:26.52:17.26:2.20:8.40:36.31。全面的结构性特征通过甲基化分析结合广泛的核磁共振(NMR)谱表明可能的支柱:→4)-β-D-GlcAp -(1→3、6)β-D-Galp -(1→3)α-L-Fucp -(1→4)-α-D-Manp -(1→2)β-D-Glcp -(1→2、4)α-D-Glcp -(1→4)-β-D-Glcp -(1→2)α-L-Fucp -(1→4)-β-D-GlcAp -(1→,多个分支段。在形态上，tvps2 -2在保持三螺旋结构的同时表现出相互连接的凸起、起伏的表面和方形/菱形晶体。此外，tvps2 -2具有良好的抗氧化活性，特别是对2,2-氮基-双（3-乙基苯并噻唑-6-磺酸）（ABTS，一半最大抑制浓度(IC50) = 0.36 mg/mL），并具有中等的1,1-二苯基-2-picrylhydrazyl （DPPH）和羟基自由基清除作用。此外，tvps2 -2对肺（A549）、肝（SK-Hep1）和结肠（HCT116和HT-29）癌细胞具有显著的抗增殖作用，其中对SK-Hep1肝癌细胞（IC50 = 1.6 mg/mL）的抑制作用最高，其作用机制是诱导凋亡、抑制迁移和调节代谢/氧化应激途径，而对健康H9C2细胞的抑制活性相对较低（2-1）。结论全面表征了一种具有抗氧化和抗肿瘤活性的新型蟾蜍毒酸性多糖（tvps2 -2），为其在抗癌药物开发中的应用提供了分子基础。图形抽象

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来源期刊

Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

6.80

自引率

3.00%

发文量

审稿时长

15 weeks

期刊介绍： Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.