基于丹参酮类化合物生源途径的丹参质量标志物研究思路探讨

Discussion on research idea of quality marker of Salvia miltiorrhiza based on biosynthetic pathway of tanshinone compounds

该研究基于中药质量标志物(Q-marker)理论以丹参为研究对象,对丹参毛状根进行诱导实验,采用实时荧光定量PCR检测毛状根中SmCPS,SmKSL,CYP76AH1 3条基因的相对表达量;分别采用UPLC和GC-MS技术检测毛状根与药材中丹参酮ⅡA、隐丹参酮、丹参酮Ⅰ、1,2-二氢丹参酮、铁锈醇以及次丹参酮二烯的含量;统计学分析表明:丹参毛状根中次丹参酮二烯与铁锈醇的含量与下游丹参酮类化合物的含量存在正相关,且丹参酮类化合物生源途径上重要基因的相对表达量可在一定程度上反映丹参酮类化合物的含量;在多省份丹参药材中,初步发现铁锈醇与丹参酮类化合物的含量也存在正相关。该研究从丹参毛状根体系可控实验入手,结合丹参药材,以丹参中特有指标性成分丹参酮类化合物的生源途径为研究基础,突出了生源途径上游基因、中游中间体化合物与下游丹参酮类化合物含量之间的重要关系,进而为从生源途径方面理解和研究丹参质量标志物(Q-marker)提供了可能的研究思路,以期建立多元、完整的中药丹参的质量控制体系。

Based on the theory of Q-marker,the hairy root of Salvia miltiorrhiza and S.miltiorrhiza in many provinces were studied.The relative expressions of SmCPS,SmKSL and CYP76AH1 genes in hairy roots were detected by real-time fluorescence quantitative PCR and the contents of tanshinoneⅡA,cryptotanshinone,tanshinoneⅠ,1,2-dihydrotanshinone,ferruginol and miltiradiene were detected by UPLC and GC-MS,respectively.Statistical analysis shows as fllows:in the hairy root of S.miltiorrhiza,the content of miltiradiene and ferruginol is positively correlated with the content of tanshinone compounds in the downstream,and the relative expression of important genes in the biosynthetic pathway of tanshinone can reflect the content of tanshinone compounds to a certain extent;in many provinces of S.miltiorrhiza,the content of ferruginol and tanshinone compounds can also be found that there is a positive correlation between the contents.Based on the biosynthetic pathway of tanshinone compounds,which is a special index component in S.miltiorrhiza,this study focused on the important relationship between the upstream gene,the middle intermediate compound and the downstream tanshinone compound content of the biosynthetic pathway,and explored the possible research ideas of improving the quality marker system of S.miltiorrhiza,and then provided the possible research ideas for understanding and studying the quality marker of traditional Chinese medicine from the biosynthetic pathway.