【作者】 罗倩怡；

【导师】 王贺祥；

【作者基本信息】 中国农业大学 ， 微生物学， 2003， 硕士

【摘要】 获得性免疫缺陷综合症(艾滋病)是一种因感染到人类免疫缺陷病毒(HIV)而导致的传染性疾病。人免疫缺陷病毒的反转录酶、蛋白酶和整合酶在病毒复制过程中起着十分重要的作用，缺一不可。目前较为有效的抗病毒化学治疗方法是反转录酶和蛋白酶抑制剂的复合用药，这些药物虽然能够对HIV有一定的抑制效果，但长期服用会对患者产生很大的毒副作用，且容易使病毒产生耐药性，所以需要寻找出毒性低且价格便宜的药物来对抗病毒。本论文测定了多种食用菌多糖复合物对反转录酶(RT)和蛋白酶(PR)的抑制作用。实验结果表明，对HIV-1 RT有抑制作用的组分主要集中在乙醇沉淀部分，即多糖复合物部分。其中以茯苓的活性最高，1mg／ml的反应浓度下抑制率达97.4％，其次是香栓菌90.6％和茶树菇84.5％。选取活性最高的茯苓菌丝体多糖复合物来作为纯化对象，经过DEAE-纤维素阴离子交换层析，CM-纤维素阳离子交换层析和FPLC Superdex 75凝胶过滤方法分离纯化后，获得一种对HIV-1 RT具有高抑制活性的糖蛋白S12，并测出其蛋白部分N末端的5个氨基酸顺序：丙氨酸—谷氨酸—天冬酰胺—甘氨酸—甘氨酸。在已测的22种食用菌多糖复合物对HIV-1 PR的抑制活性实验中，反应浓度为0.5mg／ml下对蛋白酶抑制活性最高的是香乳菇，其抑制率达80.9％。更多还原

【Abstract】 Acquired Immunodeficiency Syndrome (AIDS) is a deadly pandemic disease caused by a retrovirus known as human immunodeficiency virus type 1(HIV-1). Reverse transcriptase (RT), protease (PR), and integrase (IN) are three essential enzymes to the life cycle of the HIV. The chemotherapeutic strategies have been directed at the development of inhibitors of these HIV enzymes. One of the major obstacles to the long-term treatment of AIDS is the remarkable mutation of HIV to most, if not all, of clinically used chemotherapeutic agents. Therefore, it is an ongoing need for new, structurally diverse anti-HIV agents, which are less toxic and affordable to all AIDS patients. In this study, we firstly screened the ethanol extracts from edible mushrooms on inhibition of HIV-1 RT activity. Among the mushroom ethanol extracts examined, the extract of Poria cocos exhibited over 97.4% inhibition at Img/ml. Then this crude ethanol extract was fractionated through DEAE-cellulose anion-exchange chromatography, CM-cellulose cation-exchange chromatography, and FPLC Superdex 75 gel filtration respectively, which result in the isolation of an active polysaccharide-peptide with five amino acids of AENGG in N-terminal, named SI 2. On the other hand, we also examined these extracts’ inhibitory effects on HIV-1 PR. The results showed that Lactarius camphorates, which could inhibit HIV-1 PR by 80.9%, was the only sample exhibited over 80% inhibition on PR at 0.5mg/ml.更多还原