海洋共生菌中脑苷脂对RAW264.7细胞的抗炎作用研究  

Study on the anti-inflammatory effect of brain glycosides from marine symbiotic bacteria on RAW264.7 cell

作　　者：杨丙晔 易超 陈永轩 YANG Bingye;YI Chao;CHEN Yongxuan(Xiamen Key Laboratory of Marine Medicinal Natural Products and Cell Engineering,Xiamen Medical College,Xiamen 361023,China)

机构地区：[1]厦门医学院厦门市医用海洋天然产物与细胞工程重点实验室,福建厦门361023

出　　处：《中国海洋药物》2024年第1期17-25,共9页Chinese Journal of Marine Drugs

基　　金：厦门市科技局医疗卫生指导项目(3502Z20214ZD1328);福建省科技计划面上项目(2023J011659);大学生创新创业训练项目(X202112631064,201912631045,S202212631020)资助。

摘　　要：海洋微生物因具有生长迅速、代谢可调控、菌种易选育、可持续利用等特点成为寻找药理活性物质的新源泉。使用从海洋共附生菌中提取的脑苷脂,采用CCK8法检测其对小鼠单核巨噬细胞(RAW264.7)的毒性,建立RAW264.7细胞炎症模型,检测脑苷脂对炎症因子IL-6和IL-1β的含量变化,运用实时荧光定量PCR技术检测IL-6、IL-1β、TNF-α、MCP-1、MIP-2和INOS mRNA表达量。通过Western blotting技术检测炎症相关信号通路p-ERK1/2、My D88、NF-κB、NLRP3、p62、p-IKB和p-JNK蛋白的表达量。研究结果显示,脑苷脂可以降低IL-6、IL-1β、TNF-α、MCP-1、MIP-2和INOS炎症因子的含量和mRNA表达量,抑制p-ERK1/2、My D88、NF-κB、NLRP3、p-IKB和p-JNK蛋白的表达量。这些结果表明,脑苷脂可以通过抑制My D88/NF-κB/MAPK/NLRP3信号通路实现对脂多糖诱导的RAW264.7细胞的抗炎作用。这为脑苷脂类化合物用于治疗炎症相关疾病提供数据参考,为开发新型海洋药物奠定基础。Marine microorganisms characterized by rapid growth,adjustable metabolism,easy breeding and sustainable utilization become a new source of pharmacological active substances.In this study,brain glycosides were extracted from Marine symbiotic bacteria.CCK8 assay was used to detect cell activity of RAW264.7.After the establishment of cell inflammation model,the contents of inflammatory factors IL-6 and IL-1βin brain glycosides were detected,and the mRNA levels of IL-6,IL-1β,TNF-α,McP-1,MIP-2,INOS and IFN-βwere detected by fluorescence quantitative PCR.Meanwhile,the expression levels of inflammatory signaling pathway proteins p-ERK1/2,MyD88,NF-κB,NLRP3,P62,P-IKB and P-JNK were detected by Western Blot.The results showed that brain glycosides can decrease the contents and mRNA levels of inflammatory factors IL-6,IL-1β,TNF-α,McP-1,MIP-2,INOS and IFN-β,and inhibit the protein expression levels of p-ERK1/2,MyD88,NF-κB,NLRP3,P-IKB and P-JNK.These results suggest that the anti-inflammatory effect of brain glycosides on lipopolysaccharide induced RAW264.7 cells was realized by regulating the MyD88/NF-κB/MAPK/NLRP3 signaling pathway.This study provides a reference for the use of brain glycosides in the treatment of inflammation-related diseases and lays a foundation for the development of new Marine drugs.

关 键 词：脑苷脂 抗炎作用 炎症因子 信号通路 

分 类 号：R931.77[医药卫生—生药学]