靶细胞提取联用HPLC-TOF MS法分析丹红注射液中的活性成分

Analysis of bioactive constituents in Danhong Injection by target cell extraction combined with HPLC-TOF MS

目的应用靶细胞提取联用高效液相飞行时间质谱(HPLC-TOF MS)法分析丹红注射液中的活性成分,并对其防治疾病的分子机制做生物信息学预测。方法将小胶质细胞(BV2细胞)和丹红注射液共孵育,HPLC-TOF MS法对所得的细胞破碎液进行分析,然后挖掘靶细胞结合成分中与脑卒中相关的蛋白和信号通路。结果从BV2细胞破碎液中检测到丹参素、原儿茶醛、6-羟基红花黄色素A、迷迭香酸、丹酚酸B这5种丹红注射液中的水溶性成分,再构建靶细胞结合成分-蛋白网络,映射在Biocart信号通路中,包括Toll受体信号通路、神经营养因子、细胞凋亡、MAPK信号通路等,关联脑卒中发病机制的炎症、氧化应激方面。结论该研究初步阐释了丹红注射液与脑卒中信号通路的关联。

AIM To analyze the bioactive constituents in Danhong Injection by target cell extraction and HPLC-TOF MS and predict its molecular mechanism of prevention and treatment of disease via bioinformatics.METHODS Microglia（ BV2 cells） and Danhong Injection were incubated together,the obtained cell disruption sap was analyzed by HPLC-TOF MS. Then the proteins and signaling pathways related to stroke were mined in the constituents combined with target cells. RESULTS Five water-soluble constituents（ danshensu,protocatechuic aldehyde,6-hydroxysafflor yellow A,rosmarinic acid and salvianolic acid B） in Danhong Injection were extracted from the BV2 cell disruption sap. The constituents combined with target cells-protein network were then constructed and mapped onto Biocart pathways（ Toll-like receptor signaling pathway,neurotrophic factor,apoptosis,MAPK signaling pathways,etc.）,which were associated with inflammation and oxidative stress in the mechanism of stroke. CONCLUSION This research preliminarily demonstrates the relationship between Danhong Injection and signaling pathways related with stroke.