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[苦楝树叶粗提物和半纯化提取物的细胞毒性及抗病毒活性测定]

[Assays of cytotoxicity and antiviral activity of crude and semipurified extracts of green leaves of Melia azedarach L].

作者信息

Andrei G M, Coto C E, de Torres R A

机构信息

Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.

出版信息

Rev Argent Microbiol. 1985;17(4):187-94.

PMID:2825236
Abstract

Crude extracts from fresh green leaves of Melia azedarach L contain an antiviral factor (FAV) able to inhibit the replication of several animal viruses, e.g. Polio, VSV, HSV, FMDV, Sindbis, Junín, Pichinde and Tacaribe in Vero or BHK-21 cells. Crude preparations were subjected to different steps of purification like chromatography on Sephadex G-100 and DEAE-Sephadex. The antiviral activity of G-100 and DEAE fractions was fully conserved, whereas contaminating proteins were lost. Two types of cytotoxicity tests were performed with the different fractions. Two-fold serial dilutions of each of them were added to preformed monolayers of Vero or BHK-21 cells and cellular viability was tested. While crude extracts were toxic at low dilutions (less than or equal to 1:10), G-100 and DEAE fractions were not. The other cytotoxicity assay consisted in seeding the cells in the presence of different concentrations of each fraction. G-100 fraction affected cell growth at low dilutions (less than or equal to 1:5), while DEAE fraction did not. It should be remarked that the purification procedure rendered a partial purified DEAE fraction with an increased specific activity (antiviral activity/mg of protein). It is concluded that an antiviral factor devoid of toxicity exists in M. azedarach L extracts, which exhibited a broad spectrum of antiviral activity.

摘要

苦楝鲜绿叶的粗提物含有一种抗病毒因子(FAV),能够抑制多种动物病毒在Vero或BHK - 21细胞中的复制,例如脊髓灰质炎病毒、水泡性口炎病毒、单纯疱疹病毒、口蹄疫病毒、辛德毕斯病毒、胡宁病毒、皮钦德病毒和塔卡里贝病毒。粗制制剂经过了不同的纯化步骤,如在葡聚糖凝胶G - 100和二乙氨基乙基葡聚糖凝胶上进行层析。G - 100和二乙氨基乙基葡聚糖凝胶部分的抗病毒活性完全保留,而污染蛋白则被去除。对不同部分进行了两种细胞毒性试验。将它们各自的两倍系列稀释液加入预先形成的Vero或BHK - 21细胞单层中,并检测细胞活力。虽然粗提物在低稀释度(小于或等于1:10)时具有毒性,但G - 100和二乙氨基乙基葡聚糖凝胶部分没有。另一种细胞毒性测定方法是在存在不同浓度各部分的情况下接种细胞。G - 100部分在低稀释度(小于或等于1:5)时影响细胞生长,而二乙氨基乙基葡聚糖凝胶部分则不影响。应该指出的是,纯化过程得到了一种比活性(抗病毒活性/毫克蛋白)增加的部分纯化的二乙氨基乙基葡聚糖凝胶部分。结论是苦楝提取物中存在一种无毒的抗病毒因子,其具有广谱抗病毒活性。

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