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[摘要]
目的寻找紫芝液体深层发酵液的最佳抗肿瘤活性方法。方法采用液体深层发酵得到菌丝体,利用小鼠移植肿瘤模型追踪测试抗肿瘤活性,导向得到最佳抗肿瘤活性部位,并用苯酚-硫酸法测得其总多糖质量分数;3,5-二硝基水杨酸(DNS)法测定其还原糖质量分数,并对活性部位进行体外的细胞毒试验。结果紫芝液体深层发酵液的最佳抗肿瘤活性部位为胞内水提醇沉得到沉淀的水溶部分(样品B)含多糖88.4%,含还原糖3.15%,该部分不含蛋白质;在ig给药剂量为40.5mg/kg时,该活性部位对H22肝癌小鼠的抑瘤率为63.94%,对Lewis肺癌小鼠的抑瘤率为58.32%;该活性部位对A549、LoVo、CEM、QGY-7703等4种人肿瘤细胞的IC50值分别为160、29.28、45.06、37.38μg/mL。结论紫芝液体深层发酵液的最佳抗肿瘤活性部位为胞内水提醇沉物可溶于水部分,对小鼠移植肿瘤有较好的抑制作用,但体外细胞毒作用较弱。
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[Abstract]
Objective To study the antitumor fraction from liquid submerged fermentation of Ganoderma sinense.Methods The antitumor active fractions were extracted from the hyphostroma whichwas produced by liquid submerged fermentation and their antitumor activities were observed by micetransplant tumor model.The total polysaccharides were tested by phenol-sulphuric acid method and thecontents of reducing sugars were measured by DNS method.The cytotoxic activities were tested by MTTmethod in vitro.Results The optimum fraction was the intracellular soluble precipitation(sample B)bvwater extracting with alcohol sedimentation.The contents of total polysaccharide and reducing sugar inthe optimum active fraction from liquid submerged fermentation of G.sinense were 88.4%and 3.1 5%,respectively,but no protein.The inhibitory ratios were 63.94%and 58.32 0A on the growth of H22 andLewis tumor transplanted in mice by ig administration in the concentration of 40.5 mg/kg,respectively.The IC500n four tumor cells of A549,LoVo,CEM,and QGY一7703 were 160,29.28,45.06,and 37.38pg/mL,respectively,which suggested that the active fraction had weak cytotoxic on LoVo,CEMt andQGY一7703.Conclusion The active fraction from liquid submerged fermentation of G.sinense ispolysaccharide which has better inhibitory rate on the growth of H22 and Lewis tumor transplanted in micebut their cytotoxicities are weak.
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[基金项目]
上海市自然科学基金项目(05ZR14112);上海市科委“登山行动计划”项目中药现代化专项重点项目(06DZ19706)