多汁乳菇人工繁育及抗肿瘤活性

Artificial Cultivation of Lactarius volemus and Their Inhibitory Effect on Tumor Cells

摘要:

目的: 人工繁育了多汁乳菇,提取其子实体多糖,对其抗肿瘤活性进行研究.方法: 采用热水浸提法提取多汁乳菇子实体多糖, 取培养至对数生长期的K562细胞(细胞密度为8.0×10⁸/L)于96微孔板中,分别加入多汁乳菇多糖浓度为0,5,10,20,40,80,100 mg ·L^-1,分别培养12,24,48,72 h,每组处理重复3次,采用MTT比色法,检测多汁乳菇多糖抑制K562细胞活性,采用扫描电镜以及琼脂糖凝胶电泳方法,研究促进癌细胞凋亡.结果: 成功繁育了多汁乳菇,胡萝卜,松根能刺激菌丝生长,第3年开始出菇.多汁乳菇多糖具有一定的抗肿瘤活性,多汁乳菇多糖浓度越高,抑制作用越显著.当其浓度为100 mg ·L^-1,作用时间为72 h时,达到最大的抑制率为88.2%.多汁乳菇多糖促进K562细胞凋亡的扫描电镜以及DNA ladder进行了验证,发现多汁乳菇多糖促进癌细胞凋亡,细胞形态收缩,微绒毛显著减少,同时出现了特征性的DNA ladder,并且随着多汁乳菇多糖浓度增加,产生较多的DNA ladder,诱导了细胞凋亡.结论: 多汁乳菇可作为一种潜在的食用菌和药用菌而开发利用.

Abstract:

Objective:Artificial cultivation and anti-tumor effects of polysaccharides extracted from the fruit bodies of Lactarius volemus were studied. Method: The polysaccharides of L. volemus were extracted with boiling water. The proliferation inhibition of L. volemus polysaccharides on K562 cells was adopted by 4, 5-dimethylthiazo(-2-yl) -2, 5-diphenyl-tetrazolium bromide (MTT) method. The tumor apoptosis was investigated by scanning electron microscope(SEM) and agarose gel electrophoresis. Result: Artificial cultivation of L. volemus was achieved successfully. The growth of L. volemus mycelium was stimulated by carrots and pine roots, and the fruiting bodies were obtained in the third year. Polysaccharides of L. volemus had some anti-tumor activity which inhibition effect was strengthened significantly with increasing polysaccharides concentration. The maximum inhibition rate was reached at 88.2% with concentration of L. volemus polysaccharides at 100 mg · L^-1 after 72 h. We found L. volemus polysaccharides promoted K562 tumor cells apoptosis that cell morphology contraction, significant reduction in microvilli by SEM. Meanwhile DNA ladder was appeared by agarose gel electrophoresis, which strength enhanced with increasing concentration of L. volemus polysaccharides. Conclusion: L. volemus may be a potential source for medicinal and edible mushrooms.