丹荷颗粒治疗高脂血症质量标志物发现研究

为寻找丹荷颗粒治疗高脂血症的质量标志物,该研究基于丹荷颗粒体内成分分析结果,采用Cytoscape软件构建高脂血症-共有靶点-丹荷颗粒体内成分作用网络,利用DAVID及String数据库分别对共有靶点进行生物过程、通路富集及网络拓扑分析,明确丹荷颗粒降脂作用关键靶点及成分,再建立油酸诱导的HepG2细胞脂质堆积模型验证关键成分的降脂药效,最后结合相关成分在制剂中的含量、稳定性、特征性综合确定质量标志物。结果发现,丹荷颗粒体内成分作用于CES1、ADORA2A、ADORA1、APOB、LDLR、PPARA、PPARG、INSR、ESR2、ESR1共10个高脂血症疾病靶点,主要参与脂质代谢过程以及PPAR、cAMP信号通路,其中LDLR、PPARA、PPARG可能是关键靶点,而网络中与关键靶点直接作用的成分为白藜芦醇、柚皮素。白藜芦醇和柚皮素均可降低油酸诱导的HepG2细胞内脂滴数量,改善细胞脂质堆积严重程度,并显著降低细胞内甘油三酯含量,可能为关键成分;鉴于入体成分白藜芦醇和柚皮素来源于制剂中虎杖苷及柚皮苷,且制剂中虎杖苷及柚皮苷特征明显、含量丰富、稳定,故虎杖苷及柚皮苷可作为丹荷颗粒治疗高脂血症的质量标志物。该研究为丹荷颗粒质量标准的建立提供了合理的候选质控指标。

Study on Discovery of Quality Markers for Treatment of Hyperlipidemia with Danhe Granules

To study the quality markers of Danhe granule for treatment of hyperlipidemia,the network of“hyperlipidemia-common target-components of Danhe granules in vivo”was constructed by Cytoscape software based on the results of in vivo component analysis.To determine the key target and component,the biological process,pathway enrichment and network topology of the common target were analyzed by DAVID and String database,respectively.Then HepG2 cells induced by oleic acid(OA)were selected to establish a lipid accumulation cell model to verify the lipid-lowering efficacy of the key components.Finally,combined with the content,stability and characteristics of the related components in the preparation,the quality markers were determined.Result showed that components of Danhe granule in vivo act on 10 disease targets of hyperlipidemia,including CES1,ADORA2A,ADORA1,APOB,LDLR,PPARA,PPARG,INSR,ESR2 and ESR1,which mainly regulated the process of lipid metabolism and the signal pathways of PPAR and cAMP.It was found that LDLR,PPARA and PPARG may be the key targets for the lipid-lowering effect of Danhe granules,while the components interacted with the key targets were resveratrol and naringenin.Futhermore,resveratrol or naringenin could not only reduce the number of intracellular lipid droplets,improve the severity of lipid accumulation in HepG2 cells,but also significantly reduce the content of intracellular triglyceride which may be the key components.Considering that the resveratrol and naringenin in vivo come from polydatin and naringin in preparation,while polydatin and naringin are rich and stable in preparation with obvious characteristics,they thus could be used as the quality markers of Danhe granules in the treatment of hyperlipidemia.This study provides a reasonable candidate quality control index for the establishment of the quality standard of Danhe granules.