南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (10): 1540-1546.doi: 10.12122/j.issn.1673-4254.2021.10.13

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不同相对分子质量的黑根霉胞外多糖的生物活性

李 璋，许 晅，孟 迎，马群飞，Huma Farooque Hashmi，张鹏英，陈靠山   

  1. 山东大学生命科学学院，国家糖工程技术研究中心，山东 青岛 266237
  • 出版日期:2021-10-20 发布日期:2021-11-11

Assessment of biological activities of exopolysaccharides with different relative molecular masses extracted from Rhizopus nigricans

LI Zhang, XU Xuan, MENG Ying, MA Qunfei, HUMA Farooque Hashmi, ZHANG Pengying, CHEN Kaoshan   

  1. School of Life Science, National Glycoengineering Research Center, Shandong University, Qingdao 266237, China
  • Online:2021-10-20 Published:2021-11-11

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摘要： 目的 探讨不同相对分子质量黑根霉胞外多糖的抗氧化、抗肿瘤和免疫调节活性。方法 通过分级醇沉从黑根霉的发酵液中分离得到3种不同相对分子质量的多糖，分别命名为RPS-1，RPS-2和RPS-3，检测3种多糖对DPPH、ABTS和羟自由基的清除活性。体外培养小鼠前胃癌细胞MFC、人非小细胞肺癌细胞A549和小鼠巨噬细胞RAW 264.7，给药48 h或24h后CCK-8法检测各组细胞存活率，一氧化氮试剂盒检测RAW 264.7细胞上清液中的NO水平，流式细胞仪检测3种多糖对A549细胞凋亡率的影响。结果 RPS-1，RPS-2和RPS-3均具有较好的抗氧化活性，中等相对分子质量多糖RPS-1对DPPH和ABTS自由基的清除活性显著高于RPS-2和RPS-3（P<0.05），低相对分子质量多糖RPS-3对羟自由基的清除活性显著高于RPS-1和RPS-2（P< 0.01）。3种多糖均能够抑制MFC细胞和A549细胞的增殖，激活巨噬细胞RAW 264.7，诱导A549细胞凋亡。高相对分子质量多糖RPS-2对MFC和A549细胞增殖的抑制活性高于RPS-1和RPS-3（P>0.05），RPS-2处理后A549细胞的凋亡率显著高于RPS-1和RPS-3（P<0.05）。高相对分子质量多糖RPS-2促进RAW 264.7细胞增殖的活性显著高于RPS-1和RPS-3（P<0.05），RPS-2处理后的RAW 264.7细胞释放的NO含量显著高于RPS-1和RPS-3（P<0.01）。结论 3种不同相对分子质量的多糖均具有抗氧化、抗肿瘤和免疫调节活性，低、中相对分子质量多糖RPS-1和RPS-3具有较好的抗氧化活性，高相对分子质量多糖RPS-2具有较好的抗肿瘤和免疫调节活性。

关键词: 黑根霉胞外多糖；抗氧化；抗肿瘤；巨噬细胞

Abstract: : Objective To evaluate the antioxidant, anti-tumor and immunomodulatory activities of exopolysaccharides with different molecular masses isolated from Rhizopus nigricans. Methods Three polysaccharides with different molecular masses, namely RPS-1, RPS-2 and RPS-3, were separated from the fermentation broth of Rhizopus nigricans by fractional ethanol precipitation, and their capacity for scavenging DPPH, ABTS, and hydroxyl radicals was assessed. Cell counting kit-8 was used to analyze the changes in the viability of MFC, A549 and RAW 264.7 cells following treatments with the 3 polysaccharides; The level of nitric oxide in the supernatant of RAW 264.7 cells was detected using a nitric oxide detection kit, and the apoptosis rate of A549 cells was analyzed with flow cytometry. Results All the 3 polysaccharides had good antioxidant activities, and among them RPS-1 with a medium molecular mass exhibited the strongest scavenging capacity for DPPH and ABTS radicals (P<0.05) while RPS-3 with the lowest molecular mass had the best scavenging activity for hydroxyl radicals (P<0.01). All the 3 polysaccharides were capable of inhibiting the proliferation of MFC cells and A549 cells, activating the macrophages RAW 264.7 cells, and inducing apoptosis of A549 cells. RPS-2 with the highest molecular mass showed the strongest inhibitory effects against MFC and A549 cells (P>0.05), and RPS-2 had the strongest activity for inducing apoptosis in A549 cells (P<0.05). Compared with the other two polysaccharides, RPS-2 more strongly promoted the proliferation of RAW 264.7 cells and enhanced NO release from the cells (P<0.05). Conclusion The 3 polysaccharides all have antioxidant, anti-tumor and immunomodulatory activities, and among them RPS-1 and RPS-3 have better antioxidant activities, and RPS-2 has stronger anti-tumor and immunomodulatory activities.

Key words: exopolysaccharide from Rhizopus nigricans; antioxidant; anti-tumor; macrophages cells