香菇提取物对白血病细胞HL-60增殖的抑制作用

Antiproliferation Effect of Shiitake Mushroom(Lentinula Edodes)Exudates on HL-60(Acute Promyelocytic Leukemia)Cancer Cell Line

香菇是全球第二大人工种植最为普遍的食用菌。研究表明,香菇具有抗癌、抗氧化、抗菌及降血脂等活性,目前大多数研究集中于香菇子实体的抗癌活性。在对香菇在固体培养基上进行培养的过程中,培养基中发现菌丝分泌一种棕色物质,猜测这些棕色物质可能具有某些生物活性。本研究的目的在于探讨这些棕色物质是否对急性早幼粒细胞性白血病细胞HL-60的增殖具有抑制作用。香菇在固体培养基上培养3￣4周后,通过离心和冷冻干燥得到棕色物质,然后进行一系列的分离纯化。通过台盼蓝染色排除法测定了得到的各种组分对HL-60增殖的抑制作用,通过荧光显微镜双染色法测定了各组分对HL-60细胞凋亡的诱导作用。结果表明,作用72h后,粗提物、透析所得低分子量组分和大孔树脂分离所得低极性组分对HL-60的细胞增殖具有明显的抑制作用,IC50分别为(135±0.59),(102±0.66)和(23±0.45)μg/mL。同时,粗提物中的活性成分为低极性物质,其分子量小于10000D,并非香菇多糖。荧光显微镜双染色结果表明,作用12h后,粗提物诱导HL-60细胞的凋亡指数由32%提高至64%。结果表明,香菇提取物对于HL-60细胞的增殖具有显著的抑制作用。

Shiitake mushroom, Lentinula edodes, is the second most commonly cultivated edible mushrooms in the world. It has anticancer, antioxidant, antiviral, hypolipidaemic activities, etc. Most investigations have focused on the fruiting body for such activities. Exudates were observed producing in some strains of shiitake and suspected to have certain biological activities. Therefore, the objective was to evaluate if the exudates had any anti-proliferation activity on a cancer cell line （HL-60） . Shiitake mushroom was cultivated in the potato dextrose agar. Brownish exudates were collected after 3-week incubation, centrifuged and freeze-dried. Further fractionations were carried out. The bioactivity involving the antiproliferation activity was evaluated by trypan blue exclusion method （TBEM） and epifluorescence method （EM） at different concentrations. After 72-h incubation, the inhibition concentrations （IC50 of the original crude extract, the low molecular weight fraction （LMWF） from dialysis, and the ethanol fraction from macroporous resin column of the LMWF on the proliferation of HL-60 were 135± 0.59, 102± 0.66 and 23 ± 0.45 μg/mL, respectively. Results showed that the exudates and the fractions inhibited the proliferation of HL-60. Besides, the active components were likely low polar and their molecular weights were below 10 000 Daltons. From EM, the apoptotic indices of HL-60 were 32 ± 1.26 and 64 ± 1.06% for control and treatment, respectively established for 12-h incubation. These results show that the mushroom exudates exhibited bioactivity towards the HL-60 cancer cell line, and may be used as a potential source of functional ingredient.