Su Wenwen, Wu Leilei, Liang Qichao, Lin Xiaoyue, Xu Xiaoyi, Yu Shikai, Lin Yitong, Zhou Jiadong, Fu Yang, Gao Xiaoyan, Zhang Bo, Li Li, Li Dan, Yin Yongkui, Song Gaochen
Department of Biochemistry and Molecular Biology, Mudanjiang Medical University, Mudanjiang, China.
Collage of Pharmacology, Mudanjiang Medical University, Mudanjiang, China.
Front Pharmacol. 2022 Apr 4;13:827782. doi: 10.3389/fphar.2022.827782. eCollection 2022.
The Chinese medicinal herb D. Don has antitumour effects and is used to treat liver cancer in the clinic. polysaccharide (SBP), one of the main active components extracted from D. Don, exhibits antitumour activity. However, there is still a lack of research on the extraction optimization, structural characterization, and anti-hepatoma activity of acidic polysaccharides from D. Don. In this study, the optimal extraction conditions for SBP were determined by response surface methodology (RSM): the material-liquid ratio was 1:25, the extraction time was 2 h, and the extraction temperature was 90°C. Under these conditions, the average extraction efficiency was 3.85 ± 0.13%. Two water-soluble polysaccharides were isolated from D. Don, namely, SBP-1A and SBP-2A, these homogeneous acidic polysaccharide components with average molecular weights of 1.15 × 10 Da and 1.4 × 10 Da, respectively, were obtained at high purity. The results showed that the monosaccharide constituents of the two components were fucose, galactosamine hydrochloride, rhamnose, arabinose, glucosamine hydrochloride, galactose, glucose, xylose, and mannose; the molar ratio of these constituents in SBP-1A was 0.6:0.3:0.6:30.6:3.3:38.4:16.1:8:1.4, and that in SBP-2A was 0.6:0.5:0.8:36.3:4.4:42.7:9.2:3.6:0.7. In addition, SBP-1A and SBP-2A contained uronic acid and -glucan, and the residue on the polysaccharide was mainly pyranose. The results showed that the anti-hepatoma activity of SBP-2A was better than that of SBP-1A and SBP. In addition, SBP-2A significantly enhanced HepG2 cell death, as cell viability was decreased, and SBP-2A induced HepG2 cell apoptosis and blocked the G1 phase. This phenomenon was coupled with the upregulated expression of P53 and Bax/Bcl-2 ratio, as well as the downregulated expression of the cell cycle-regulating protein cyclinD1, CDK4, and Bcl-2 in this study. Further analysis showed that 50 mg/kg SBP-2A inhibited the tumour growth in H22 tumour-bearing mice, with an average inhibition rate of 40.33%. Taken together, SBP-2A, isolated and purified from showed good antitumour activity and , and SBP-2A may be a candidate drug for further evaluation in cancer prevention. This study provides insight for further research on the molecular mechanism of the anti-hepatoma activity of polysaccharide.
中药密花豆具有抗肿瘤作用，临床上用于治疗肝癌。密花豆中提取的主要活性成分之一多糖（SBP）具有抗肿瘤活性。然而，关于密花豆酸性多糖的提取优化、结构表征及抗肝癌活性仍缺乏研究。本研究采用响应面法（RSM）确定了SBP的最佳提取条件：料液比为1:25，提取时间为2小时，提取温度为90℃。在此条件下，平均提取效率为3.85±0.13%。从密花豆中分离出两种水溶性多糖，即SBP-1A和SBP-2A，获得了高纯度的这些均一的酸性多糖成分，其平均分子量分别为1.15×10⁵Da和1.4×10⁵Da。结果表明，这两种成分的单糖组成有岩藻糖、盐酸半乳糖胺、鼠李糖、阿拉伯糖、盐酸葡萄糖胺、半乳糖、葡萄糖、木糖和甘露糖；这些成分在SBP-1A中的摩尔比为0.6:0.3:0.6:30.6:3.3:38.4:16.1:8:1.4，在SBP-2A中的摩尔比为0.6:0.5:0.8:36.3:4.4:42.7:9.2:3.6:0.7。此外，SBP-1A和SBP-2A含有糖醛酸和β-葡聚糖，多糖上的残基主要为吡喃糖。结果表明，SBP-2A的抗肝癌活性优于SBP-1A和SBP。此外，SBP-2A显著增强HepG2细胞死亡，细胞活力降低，SBP-2A诱导HepG2细胞凋亡并阻滞G1期。本研究中，这种现象伴随着P53表达上调以及Bax/Bcl-2比值升高，同时细胞周期调节蛋白cyclinD1、CDK4和Bcl-2表达下调。进一步分析表明，50mg/kg SBP-2A可抑制H22荷瘤小鼠肿瘤生长，平均抑制率为40.33%。综上所述，从密花豆中分离纯化得到的SBP-2A具有良好的抗肿瘤活性，SBP-2A可能是一种有待进一步评估用于癌症预防的候选药物。本研究为深入研究密花豆多糖抗肝癌活性的分子机制提供了思路。
