Screening active fractions from Pinus massoniana pollen for inhibiting ALV-J replication and their structure activity relationship investigation
Screening active fractions from Pinus massoniana pollen for inhibiting ALV-J replication and their structure activity relationship investigation
复制标题
马尾松花粉抑制ALV-J复制活性组分的筛选及其构效关系研究
DOI：
10.1016/j.vetmic.2020.108908
复制
发表时间：
2021-01-01
影响因子：
3.3
通讯作者：
Zhu, Ruiliang
中科院分区：
文献类型：
--
作者：
Cui, Wenping;Huang, Jin;Zhu, Ruiliang
目的是鉴定抗 ALV-J 复制的多糖活性组分并阐明它们的结构活性关系。最佳提取条件为提取温度90℃、pH 9、液固比30:1。在此条件下，总多糖的提取率为6.5%+/-0.19%。然后通过DEAE-52纤维素和Sephadex G-200凝胶纯化总多糖。三个级分 PPP-1、PPP-2 和 PPP-3 的分子量分别为 463.70、99.41 和 26.97 kDa。三种多糖组分均由10种不同比例的单糖组成。与主要由葡萄糖组成的PPP-1相比，PPP-2和PPP-3含有更高比例的半乳糖、葡萄糖醛酸和半乳糖醛酸。刚果红测定表明PPP-2可能具有三螺旋结构，而PPP-1和PPP-3不存在。体外实验表明，800 μg·mL(-1)浓度下，多糖组分无明显细胞毒性(P > 0.05)。抗病毒试验显示PPP-2活性最强，表明PPP-2是主要的抗病毒成分。构效关系表明，多糖组分的抗病毒活性受其单糖组成、分子量和三螺旋结构的影响，是多种分子结构因素综合作用的结果。这些结果表明PPP-2可以作为替代合成抗病毒药物的有价值的产品，并为马尾松花粉多糖作为抗病毒药物的有用来源的未来应用提供支持。
The objective was to identify the active fractions of polysaccharide against replication of ALV-J and elucidate their structure activity relationship. The optimal extraction conditions were extracting temperature 90 degrees C, pH 9 and the ratio of liquid to solid 30:1. Under these conditions, extraction yield of total polysaccharide was 6.5 % +/- 0.19 %. Total polysaccharide was then purified by DEAE-52 cellulose and Sephadex G-200 gel. Three fractions, PPP-1, PPP-2, and PPP-3, were identified with molecular weight of 463.70, 99.41, and 26.97 kDa, respectively. Three polysaccharide fractions were all composed of 10 monosaccharides in different proportions. Compared with PPP-1, which was mainly composed of glucose, PPP-2 and PPP-3 contained a higher proportion of galactose, glucuronic acid and galacturonic acid. The Congo red assay indicated that the PPP-2 may have a triple helical structure, while PPP-1 and PPP-3 were absent. In vitro assay showed that there was no significant cytotoxicity among the polysaccharide fractions under the concentration of 800 mu g mL(-1) (P > 0.05). The antiviral test showed that PPP-2 had the strongest activity, indicating PPP-2 was the major antiviral component. The structure-activity relationship showed that the antiviral activities of polysaccharide fractions were affected by their monosaccharide composition, molecular weight, and triple helical structure, which was a result of a combination of multiple molecular structural factors. These results showed that the PPP-2 could be exploited as a valued product for replacing synthetic antiviral drugs, and provided support for future applications of polysaccharide from Pinus massoniana pollen as a useful source for antiviral agent.