【作者】 李明玉；

【导师】 台桂花；

【作者基本信息】 东北师范大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 胶陀螺( Bulgaria inquinans Fr. )属子囊菌纲(Ascomycetes) ,蜡钉菌目(Helotiales) ,胶陀螺科(Bulgariaceae) ,胶鼓菌属( Bulgaria Fr. )真菌。胶陀螺富含各种营养成分,是东北山区人们习食的一种真菌。沸水煮提胶陀螺子实体,过滤除去残渣,浓缩滤液,经醇沉,常规干燥得到胶陀螺粗多糖BP;BP水溶液进一步离心除去粗多糖中的杂质,醇沉精制,沉淀冰冻干燥得到胶陀螺多糖BP1;BP1水溶液经反复冻融除杂,Sevag法脱蛋白,最后得到较均一的胶陀螺多糖组分BP2。我们通过柱层析分别分析了三者的分子量分布情况;由高效液相层析测定了它们各自的单糖组成和比例;并测定了BP、BP1、BP2中的蛋白质和糖的含量;还对它们的一些理化性质进行了研究。以ICR系雄性小白鼠为实验材料,初步探究了BP对移植性肉瘤S-180的抑制作用。每组实验动物连续灌胃给药10天,停药后测量小鼠的体重和小鼠肿瘤、胸腺、脾脏的重量。计算肿瘤抑制率,胸腺指数,脾脏指数。结果表明:BP单独给药,对小鼠移植性肉瘤S-180的抑制作用不明显,测得肿瘤抑制率为22.7%,但BP与抗肿瘤药物5-氟尿嘧啶(5-Fu)联合使用,可以显著提高5-氟尿嘧啶的药效,使得肿瘤抑制率由单独使用5-Fu的36.8%提高至46.5%。同时BP可降低5-氟尿嘧啶对小鼠机体造成的副作用,减小其对小鼠免疫系统的不良影响。为进一步探讨胶陀螺粗多糖BP对ICR小鼠免疫调节能力的影响,探寻其增强5-Fu抗肿瘤药效的原因。我们进行了小鼠体内淋巴细胞增殖试验,结果表明:BP对淋巴细胞增殖有促进作用,尤其对脂多糖诱导的B细胞增殖能力的促进作用更加显著,并呈剂量依赖关系,推测BP可能是一种B细胞丝裂原。由此可见,胶陀螺粗多糖对抗肿瘤药物5-氟尿嘧啶的增效作用,可能是通过BP激活小鼠机体内相关的免疫调节途径实现的。更多还原

【Abstract】 A crude water-soluble polysaccharide BP was extracted from Bulgaria inquinans Fr. Further purification by ethanol precipitation and lyophilization resuled in a fraction BP1. After deproteination by Sevage method and freeze thawing, fraction BP2 was obtained. The content of protein and sugar of BP, BP1 and BP2 was determined. The molecular weight distribution of these three polysaccharide fractions was analyzed by column chromatography.The antitumor activity of Bulgaria inquinans polysaccharides against Sarcoma 180 was tested in ICR mice. After successive administered for ten days, the weight of body, tumor, spleen and thymus was measured and tumor-inhibition ratio was examined. BP showed some inhibitory effect on the S-180 alone, but the effect was not significant. Combined treatment with the anti-tumor medicine 5-fluorouracil (5-Fu) and BP, however, resulted in more significant effect than the effect of 5-Fu alone. BP also restored the weight of spleen that suppressed by 5-Fu. These results suggested that clinical trials of BP as an adjuvant in cancer chemotherapy of 5-Fu are highly feasible.Lymphocyte proliferation activity of the crude polysaccharide BP in mice was determined using MTT assay. Results showed that lymphocyte proliferation activity of BP- treated mice was significant higher than the control, especially for LPS-induced B cell, and exhibited a dose-dependent manner. The result indicated that the anti-tumor activity of BP polysaccharide may be mediated by its immunomodulating activity.更多还原