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题名: 若干天然产物抗病毒活性及其作用机制的研究
作者: 李艳梅
学位类别: 博士
答辩日期: 2006-05-30
授予单位: 中国科学院昆明植物研究所
授予地点: 昆明植物研究所
导师: 郝小江
关键词: 马蓝 ; 抗病毒活性 ; 植物病毒 ; 烟草花叶病毒 ; 正链RNA病毒
学位专业: 植物学
中文摘要: 首次发现双裂孕烷甾体glaucogenin C(1)及其单糖苷cynatratoside A (2)、以及徐长卿(Cynanchum paniculatum)中glaucogenin C(1)的3个五糖苷3-5具有广谱的抗-正链RNA病毒作用,如辛德比斯病毒(SINV), 西方马脑炎病毒(EEEV)和盖塔病毒(GETV)等。该类化合物对其它RNA病毒或DNA病毒没有作用,且对病毒宿主没有毒性。体内实验证实:该类化合物能保护BALB/c小鼠不被SINV侵染致死而对小鼠没有副作用。以TMV和SINV作为模式病毒,探讨了该类化合物抗病毒的作用机制,结果表明:该类化合物选择性地抑制TMV亚基因组RNA的合成而对病毒基因组的复制没有影响。该具有高识别性的作用机制为首次发现。根据该研究结果,我们认为:影响病毒的亚基因组RNA的合成可以作为发现抗病毒活性成分的新靶点,对于复制周期中产生亚基因组RNA的-正链RNA病毒而言,双裂孕烷甾体glaucogenin C(1)及其苷有可能成为新的一类抑制剂。 Isatin 类似物能抑制TMV对烟草的感染、提高植物体内水杨酸(SA)的积累、显著PR-1基因的表达、提高苯丙氨解氨酶的活性。化合物7诱导的5个蛋白与植物防御响应中水杨酸诱导的蛋白相同,而另有2个抗病毒蛋白为化合物7独立诱导。蛋白质组分析表明,由化合物7诱导的蛋白磷酸化也与水杨酸诱导的相同。表明该化合物是植物防卫反应的外源信号分子,其激活的植物防卫反应依赖于SA途径。由三帖皂苷Cinchonaglycoside C(8)处理后的烟叶中分离得到一个31 kDa的具有明显抗TMV活性的核糖体失活蛋白(RIP),由其蛋白序列可知其为一新的单链RIP蛋白,不同于已报道的其它植物核糖体失活蛋白,但其N端又表现出与别的RIP有一定的同源性。该蛋白在ng/ml级浓度时能直接破坏TMV外壳蛋白(CP)的合成从而阻止病毒的长距离移动和病毒的增殖,对植物的正常生长没有影响。研究表明:该蛋白只有在Cinchonaglycoside C(8)存在的条件下表达,与SA信号途径无关。
英文摘要: We unprecedentedly provide evidence that a new class of compounds, seco-pregnane steroid glaucogenin C(1) and its monosugar-glycoside cynatratoside A (2)of Strobilanthes cusia and three pantasugar-glycosides 3-5 of glaucogenin C of Cynanchum paniculatum, are effective and selective inhibitors of a broad spectrum of alphavirus-like positive-strand RNA viruses including TMV, SINV, EEEV and GETV, but not other RNA or DNA viruses, yet they were not toxic to host cells. In vivo administration of the compounds protected BALB/c mice from lethal SINV infection without adverse effects on the mice. Using TMV and SINV as models, studies on the action mechanism revealed that the compounds predominantly inhibited the synthesis of viral subgenomic (sg) RNA(s) without affecting the accumulation of viral genomic RNA, implying that the compounds selectively target the viral sgRNA synthesis machinery. Our work suggested that the viral sgRNA synthesis machinery could be a new target for the discovery of antiviral drugs, and that seco-pregnane steroid and its four glycosides found in the two medicinal herbs have the potential to control infections by positive-strand RNA viruses which make sgRNA(s) during their replication cycle. Isatin ramification activated plant resistance to TMV infection is mainly dependent and requires SA mediated signaling pathways as well as one or more unidentified factors activated by the compound. Cinchonaglycoside C inhibit TMV multiplication by induced A new single-chained ribosome-inactivating protein (RIP) with antiviral activity. This protein can directly impair synthesis of coat protein (CP) of TMV, resulting in inhibition of TMV long distance movement and multiplication. This protein was synthesized only in the present of Cinchonaglycoside C (1) and independent of salicylic acid (SA) signal pathway.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.kib.ac.cn/handle/151853/40
Appears in Collections:昆明植物所硕博研究生毕业学位论文_学位论文

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Recommended Citation:
若干天然产物抗病毒活性及其作用机制的研究.李艳梅[d].中国科学院昆明植物研究所,2006.20-25
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