| The quality standard for Chinese Materia Medica(CMM)is the key issues restricting its scientific development.At present,the existing standards are monotonous and biased,separated from the clinic.Chrysanthemum,as both medicine and food,is widely used throughout the world.However,since it has many species,complicated ingredients,and can be easily influenced by the growth environment,place of origin,germplasm resources,harvest time,all affect the changes of its chemical composition.In order to promote the development of quality evaluation system of CMM targeting the clinic,this research made connecting ingredients with efficacy as the breakthrough point,and conducted research on quality evaluation of Chrysanthemi Flos based on efficacy-component correlation analysis.The purpose of this study is to establish a new standard for evaluating the quality of Chrysanthemi Flos based on the correlation between pharmacological activity and ingredients,build a multi-level evaluation and innovation system that objectively reflects the quality of Chinese herbal medicine,so as to promote its quality standards transforming to more adaptable manner and to ensure that the quality of Chrysanthemi Flos is controllable,clinically effective,and the products made of Chrysanthemi Flos are high quality,providing strong scientific and technological support for the high-quality development of Chinese herbal medicine industry.Under the guidance of TCM theory,this research is based on modern information network technology,computer-aided technology,chemical analysis technology,and molecular biology technology,taking Chrysanthemi Flos as the research object,comprehensively evaluate the quality of Chrysanthemi Flos from the aspects of network pharmacology,virtual screening,morphological traits,plant metabolism,chemical analysis,and pharmacodynamic activity,adopt molecular docking technology,UPLC-Q-TOF-MS analysis,PCR and other technologies.With the help of chemometric methods such as medical statistics,fingerprint similarity,principal ingredient analysis,cluster analysis,and heat map analysis,to establish the quality evaluation standard base on "efficacy-component "correlation of Chrysanthemi Flos.The research started with Chrysanthemi Flos ingredients screening based on medicinal materials and chemical biomarkers.to conduct metabonomics of plants research,foucs on qualitative and quantitative analysis on 27 species of fresh Chrysanthemi Flos in three flowering seasons.in order to found the stable and controllable compouds in Chrysanthemi Flos with the changes of species,flower sea-sons and chemical ingredients.Secondly,screened the Chrysanthemi Flos target ingredients based on the chemical and pharmacological biomarkers:To analyze the content of 13 ingredients of 52 Chrysanthemi Flos extracts;make comparative analysis on different species,different flower seasons and different extract methods;evaluate the pharmacodynamic activity and screen the quality control markers.Network pharmacology and molecular docking technology are used to quality markers screening first time in this article.Then took in vitro cell inflammation model to evaluate the differences activity of different varieties of Chrysanthemi Flos,with detecting cell viability and nitric oxide concentration,as well as expression levels of TNF-?,IL-6,IL-1?,iNOS,COX-2,and PGE2 mRNA,so as to study the correlation between chemical biomakers and anti-inflammatory effects and determine the effective active compouds in Chrysanthemi Flos.Finally,by establishing a new method based on efficacy-component correlation on quality evaluation standards of Chrysanthemi Flos,study in the grade evaluation of 50 kinds of Chrysanthemi Flos based on the quality constant method.Chemical-based study shows that 73 compounds have been detected in Chrysanthemi Flos with metabolomics method.Caffeoquinic acid and flavonoids are the main chemical ingredients,and the quality markers as the evaluation standard of Chrysanthemi Flos can be selected from these two categories.Some compounds are commonly seen in Chrysanthemi Flos with high quantity,and the amount are stable among different species,such as 4,5-DCQA,which is expected to become a quantitative compound;some compounds are high in all samples and the differences between species are small,such as 3-CQA is suitable for qualitative identification.some compounds with low content but big dispersion in all species are not suitable for being the quality evaluation markers of Chrysanthemi Flos.The changing of the amount of each comound in flowering period is much smaller than the differences between species.This research detected 13 main compounds in Chrysanthemi Flos extracts,10 compounds were tested in all kinds of Chrysanthemi Flos extracts.Some of the compounds had the highest content and a large degree of dispersion,which could be used to evaluate the quality of Chrysanthemi Flos or identify the difference between varieties,like 3,5-DCQA.The content of some compounds are high in all samples with small differences between varieties,like 3-CQA,which is suitable for qualitative identification markers.The analysis on species shows that Gongju is of the best quality.The classification of Chrysanthemi Flos extracts based on chemical ingredients is basically the same as the traditional Chrysanthemi Flos classification.The analysis on the harvesting period found that the chemical composition of the bud and the full-blossom of Chrysanthemi Flos was the same,but the content of most compounds in the bud was higher than that of the full-blossom Chrysanthemi Flos.In vitro anti-inflammatory test showed that the most important anti-inflammatory pathway of Chrysanthemi Flos was PTGS.Virtual molecular screening showed that Lut-7-O-(3-G etc.has the strongest binding ability with inflammatory target proteins.The main chemical compounds of Chrysanthemi Flos have the strongest potential for regulating inflammatory factors COX-2 and iNOS.Chrysanthemi Flos has obvious inhibitory effects on inflammatory mediators and inflammatory factors.There are significant differences in the anti-inflammatory effects of different Chrysanthemi Flos species.The correlation analysis results showed that Lut-7-O-?-G etc.had a good correlation with anti-inflammatory effects,and could be used a as quality evaluation markers.The target compounds as quality evaluation markers were screened base on"efficacy-component " correlation of Chrysanthemi Flos determines that Lut-7-O-?-G,3-CQA,3,5-DCQA,4,5-DCQA etc.could be the chemical quality markers for Chrysanthemi Flos quality evaluation,which has a clearer relationship between ingredients and efficacy compared with the existing chemical quality markers in the Chinese Pharmacopoeia.the screened chemical quality markers is more scientific and refined.It is an optimization and innovation of Chrysanthemi Flos quality evaluation based on the Chinese pharmacopoeia.Using this method,this study tested 50 types of Chrysanthemi Flos which were found in the market,using the method of grade evaluation based on the quality constant.this study will play a positive role in regulating the market,improving the quality of Chrysanthemi Flos,guiding market prices,and ensuring the safety and effective use in clinic. |
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