| 摘要: |
| 【目的】对1株野外采集的蝉虫草进行分离和鉴定，分析比较其发酵菌丝体胞内和胞外多糖的抗肝癌细胞HepG-2活性及多糖的组成。【方法】通过形态学特征和ITS序列分析，对供试菌株进行鉴定；采用MTT法、细胞划痕试验，比较供试菌株菌丝体胞内和胞外多糖对人肝癌细胞HepG-2的抑制活性以及对人正常肝细胞LO2的毒性，并通过高效液相色谱法和凝胶渗透色谱法，比较分析2种多糖的成分及分子质量分布。【结果】根据菌株形态学特征和ITS序列分析，供试菌株为蝉虫草Cordyceps cicadae；其胞内和胞外多糖均可极显著抑制HepG-2细胞活性（P<0.01），半数抑制浓度（IC50）值分别为3.23和0.45 mg/mL，细胞迁移率分别为38.72%和24.79%，提示胞外多糖比胞内多糖具有更强的抗肝癌细胞HepG-2活性；且2种多糖对人LO2细胞均无明显毒性；进一步分析发现，胞外多糖中半乳糖醛酸的含量是胞内多糖的23.53倍，其分子质量的94.1%集中在19.1～85.0 ku，胞内多糖中核糖是其特征性单糖，高分子质量多糖（>250.0 ku）分布占比明显高于胞外多糖。【结论】野生蝉虫草菌株发酵所产胞外多糖的抗肝癌细胞HepG-2活性较胞内多糖强，二者在单糖组成和分子质量分布上存在明显差异。 |
| 关键词: 蝉虫草 分离与鉴定 胞内胞外多糖 抗肝癌活性 |
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| 基金项目:国家重点研发计划项目（2018YFD0400200）；福建农林大学现代种业工程研究院建设经费项目 |
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| Identification of a wild Cordyceps cicadae strain and comparison of its intracellular and extracellular polysaccharides in anti-hepatoma activity |
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LIU Chengyi,HAO Xinyi,LIN Peilin,et al
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| Abstract: |
| 【Objective】A Cordyceps cicadae strain was isolated and identified in the wild and the anti hepatoma activity and components of its intracellular and extracellular polysaccharides were analyzed.【Method】The wild Cordyceps cicadae was identified by morphological characteristics and ITS sequences.Inhibitory activity on human hepatocell carcinoma cell HepG-2 and toxicity on human normal cell LO2 of intracellular and extracellular polysaccharides from Cordyceps cicadae were compared by MTT assay and wound healing assay.The components of intracellular and extracellular polysaccharides were analyzed by HPLC and Gel Permeation Chromatography.【Result】The wild strain was identified as Cordyceps cicadae.Its intracellular and extracellular polysaccharides significantly inhibited the growth of HepG-2 cells (P<0.01) with IC50 of 3.23 and 0.45 mg/mL and cell migration rates of 38.72% and 24.79%,respectively.It indicated that extracellular polysaccharides possessed stronger anti-HepG-2 activity than intracellular polysaccharides.Both intercellular and extracellular polysaccharides had no obviously effects in LO2 cells.Further analysis showed that the content of galacturonic acid in extracellular polysaccharides was 23.53 times that in intracellular polysaccharides,and the molecular weight of extracellular polysaccharides was mainly at 19.1-85.0 ku (94.1%).Ribose only existed in intracellular polysaccharides.Relative contribution of high molecular weight polysaccharides in intracellular polysaccharide (>250.0 ku) was higher than in extracellular polysaccharides.【Conclusion】The extracellular polysaccharides extracted from fermentation of wild Cordyceps cicadae strain showed stronger anti-HepG-2 activity than intracellular polysaccharides,and there were significant differences in composition and molecular weight distribution of intracellular and extracellular polysaccharides. |
| Key words: Cordyceps cicadae isolation and identification intracellular and extracellular polysaccharides anti-hepatoma activity |
