金环胡蜂虫草来源短密青霉(Penicillium brevicompactum) VMFST24次生代谢产物的分离鉴定及抗炎活性评价

Isolation, identification, and anti-inflammatory activity evaluation of secondary metabolites of Penicillium brevicompactum VMFST24 from Vespa mandarinia Cordyceps sensu lato

摘要:

【背景】昆虫是生物种类最多的类群，是地球上尚未被充分开发与利用的巨大生物资源，昆虫来源真菌能产生结构丰富的次生代谢产物，是活性天然产物的重要来源。【目的】以金环胡蜂虫草来源短密青霉属(Penicillium brevicompactum) VMFST24为研究对象，分离鉴定其次级代谢产物并进行抗炎活性测试。【方法】首先以脂多糖(lipopolysaccharide, LPS)诱导的RAW264.7细胞炎症模型对P. brevicompactum VMFST24的水萃取物、正丁醇萃取物、乙酸乙酯萃取物进行抗炎活性检测。再以活性为导向，采用多种色谱技术对P. brevicompactum VMFST24乙酸乙酯萃取的中等极性部位进行分离纯化，获得单体化合物进行抗炎活性测试；最后采用气相色谱-质谱(gas chromatography-mass spectrometry, GC-MS)技术对小极性部位的化学成分进行检测分析。【结果】通过抗炎活性检测发现P. brevicompactum VMFST24发酵产物的乙酸乙酯萃取物具有较好活性，对其进行分离纯化，共鉴定5个化合物：mycophenolic methyl ester (1)、α-acetylorcinol (2)、3-methoxy-6,8-dihydroxy-3-methyl-3,4-dihydroisocoumarin (3)、3,5-二羟基苯甲酸甲酯(4)和苔色酸(5)，其中化合物2和4具有抗炎活性；通过GC-MS技术对P. brevicompactum VMFST24乙酸乙酯萃取物的小极性部位化学成分进行检测分析，结果表明其富含大量的脂肪酸及其酯和酚类成分。【结论】金环胡蜂虫草来源真菌Penicillium brevicompactum具有产生抗炎活性次生代谢产物的能力，本研究为胡蜂虫草真菌活性天然药物的开发奠定了基础。

Abstract:

[Background] Insects represent the group with the highest number of biological species and constitute a vast biological resource on earth that has yet to be fully explored and utilized. Fungi derived from insects can produce structurally diverse secondary metabolites and serve as a significant source of active natural products. [Objective] To isolate and identify the secondary metabolites of the fungus Penicillium brevicompactum VMFST24 derived from Vespa mandarinia Cordyceps sensu lato and evaluated their anti-inflammatory activities. [Methods] First, the RAW264.7 cell model of inflammation induced by lipopolysaccharide was employed to evaluate the anti-inflammatory activities of the water extract, n-butanol extract, and ethyl acetate extract of P. brevicompactum VMFST24. Subsequently, based on the results of the activity tests, the moderately polar fraction of the ethyl acetate extract was isolated and purified by multiple chromatographic techniques. The obtained monomeric compounds were then tested for their anti-inflammatory activities. Finally, GC-MS was utilized to detect and analyze the chemical components of the low polar fraction. [Results] The ethyl acetate extract of the fermentation product of P. brevicompactum VMFST24 exhibited a significant anti-inflammatory activity. Five compounds were identified from this extract, including mycophenolic methyl ester (1), α-acetylorcinol (2), 3-methoxy-6,8-dihydroxy-3-methyl-3,4- dihydroisocoumarin (3), methyl 3,5-dihydroxybenzoate (4), and o-orsellinic acid (5). Among them, compounds 2 and 4 demonstrated anti-inflammatory activities. Additionally, GC-MS analysis of the low polar fraction of the ethyl acetate extract revealed high abundance of fatty acids, their esters, and phenols. [Conclusion] P. brevicompactum, a fungus associated with Vespa mandarinia Cordyceps sensu lato, has the capacity to produce anti-inflammatory secondary metabolites. This research establishes a foundation for the development of active natural drugs derived from Cordyceps fungi associated with Vespa.