基于NF-κB信号通路筛选产抗炎活性次级代谢产物的红树林真菌

摘要：目的 以福建漳江口红树林沉积物来源真菌为对象，筛选产抗炎活性次级代谢产物的红树林真菌。方法 将pNFκB-luc 与pRL-SV40-C报告基因质粒共转染Raw264.7细胞，脂多糖(LPS)刺激该细胞建立体外炎症模型，构建双荧光素酶报告基因筛选体 系，对福建漳江口红树林沉积物来源真菌的发酵粗提物进行抗炎活性筛选；CCK-8法检测粗提物对Raw264.7细胞活力影响；光学显 微镜观察粗提物对LPS刺激的Raw264.7细胞分化作用。结果 从102株红树林沉积物来源真菌中获得5株阳性菌株，其发酵粗提物可 明显抑制LPS诱导的Raw264.7细胞中NF-κB信号通路活化，菌株编号分别为H788、H1458、H515、XT20-8和T2-3'。结论 5株红树 林沉积物来源真菌产生的次级代谢产物可通过抑制NF-κB信号通路发挥抗炎作用，其活性成分值得开展进一步化学研究。

Screening of mangrove fungi producing anti-inflammatory secondary metabolites based on NF-κB signaling pathway

Abstract Objective To screen fungi producing anti-inflammatory secondary metabolites isolated from sedimentss collected in Zhangjiang River Estuary Mangrove National Nature Reserves, Fujian province. Methods The pNFκB-luc and pRL-SV40-C reporter gene plasmids were co-transfected with Raw264.7 cells, which were stimulated by lipopolysaccharide (LPS) to establish an in vitro inflammation model, and a dual-luciferase reporter gene screening system was constructed to screen the anti-inflammatory crude extracts produced by mangrove sediment-derived fungi; effect of crude extracts on Raw264.7 cells viability was measured using CCK-8 assay;the differentiation effect of crude extracts on LPS-stimulated Raw264.7 cells was observed by optical microscope. Results 5 positive strains were obtained from 102 mangrove sediment-derived fungi, and their fermented crude extracts significantly inhibited NF-κB signaling pathway activation in LPS-stimulated Raw264.7 cells, strain codes were H788, H1458, H515, XT20-8 and T2-3'. Conclusion The secondary metabolites produced by 5 mangrove sediment-derived fungi strains exert anti-inflammatory effects by inhibiting NF-κB signaling pathway, and these active components deserved studying by natural products