Study on mechanism of hepatoprotective effect of Chrysanthemum morifolium Ramat. based on metabolomics with network analysis and network pharmacology.

Hangju (HJ), the dried flower heads of Chrysanthemum morifolium Ramat., has a significant hepatoprotective effect. However, its underlying protection mechanism against acute liver injury (ALI) has been unclear. An integrated strategy based on metabolomics with network analysis and network pharmacology was developed to explore the potential molecular mechanism of HJ on ALI protection. Firstly, differential endogenous metabolites were screened and identified by metabolomics approach and metabolic pathway analysis was performed by MetaboAnalyst. Secondly, marker metabolites were used to construct metabolite-response-enzyme-gene networks and discover hub metabolites and potential gene targets in network analysis. Thirdly, hub genes through the protein-protein interaction (PPI) network were acquired by the aid of network pharmacology. Finally, the gene targets were taken to intersect with the relevant active ingredients for validation by molecular docking. In total, 48 flavonoids were identified in HJ, which were associated with 8 potential therapeutic targets in network pharmacological analysis. Biochemistry and histopathology analysis demonstrated that HJ exerted hepatoprotective effects. 28 biomarkers were successfully identified as possible biomarkers for the prevention of ALI. The sphingolipid metabolic pathway and the glycerophospholipid metabolic pathway was considered a crucial signaling pathway by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. In addition, phosphatidylcholine and sphingomyelin were considered as hub metabolites. Twelve enzymes and 38 genes were considered as potential targets in the network analysis. Based on the combined analysis above, HJ was shown to modulate 2 key upstream targets, including PLA2G2A and PLA2G4A. Molecular docking showed that active compounds of HJ had high binding affinity with these key targets. In conclusion, the flavonoid components of HJ can inhibit PLA2 and regulate glycerophospholipid and sphingolipid metabolism pathway to delay the pathological process of ALI, which may be a potential mechanism of HJ against ALI.

Investigation of pharmacodynamic material basis of Anemarrhenae Rhizoma and its processed products based on plant metabolomics and molecular docking technology.

RATIONALE: Anemarrhenae Rhizoma (AR) has been an often used traditional Chinese medicine (TCM) for a long time. Its salt-processed form is one of the most common application forms. Modern pharmacological research has shown that the salt-processed product has various significantly enhanced pharmacological activities. However, the pharmacodynamic material basis of this change is not yet known. The aim of this study was to develop a strategy to screen pharmacodynamic substances in AR and salt-processed AR (SAR). METHODS: An integrated strategy combining plant metabolomics with molecular docking technology was established to screen pharmacodynamic substances. The plant metabolomics analysis was performed to select the chemical markers between AR and SAR. Then, molecular docking technology was applied to explore the relationship between chemical markers and diabetes targets (α-glucosidase). Finally, potential quality control markers were screened. RESULTS: There were significant differences in the quantification of nine steroidal saponins between AR and SAR. The results of plant metabolomics analysis showed a quite clear discrimination including 29 chemical markers between AR and SAR. Taking the hypoglycemic activity into consideration, 16 steroidal saponins were selected as potential quality markers. CONCLUSIONS: The developed method not only supplied an optional solution to search for pharmacophores in AR and SAR, but also provided a foundation for the study of the differential components and pharmacodynamics in various processed products of TCMs.

A holistic comparison of flavor signature and chemical profile in different harvesting periods of Chrysanthemum morifolium Ramat. based on metabolomics combined with bioinformatics and molecular docking strategy.

Taiju and Duoju are products of Hangbaiju (HJ) obtained during different collection periods, and they have been commonly used as ingredients in tea beverages and dietary traditional Chinese medicine. This study reports an integrated strategy based on metabolomics, bioinformatics and molecular docking to further explore the effect of the harvesting period on the metabolic profile and clinical efficacy of HJ. Firstly, gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) were employed for non-targeted metabolomics profiling of essential oils and flavonoids. A sequential window acquisition of all theoretical fragment-ion spectra information-dependent acquisition (SWATH-IDA) bi-directionally verified (SIBDV) method was developed that integrates the advantages of both SWATH and IDA in characterizing flavonoids. Chemometric methods were then used to screen potential chemical markers. Furthermore, HJ is effective in hepatoprotective functions. Therefore, hepatocellular-carcinoma-related differentially expressed genes were obtained using bioinformatics, and the corresponding proteins were molecularly docked with diagnostic chemical markers. In total, 78 volatile oils and 63 flavonoids were tentatively identified. The results allowed the selection of 11 metabolites (5 volatile oils and 6 flavonoids), which are nominated as novel markers for material authentication of Taiju and Duoju. Additionally, two proteins associated with hepatoma were screened using bioinformatics. All six flavonoid markers with binding energies of <-5 kcal mol-1 were considered to be anti-hepatoma biomarkers. Noticeably, in Taiju, the content of hydroxygenkwanin showed a downward trend, but the content of the other five flavonoids and the five flavored volatile difference compounds had an upward trend. This bestows a unique flavor profile on Taiju, leading to differences in sensory aroma and clinical efficacy in Taiju and Duoju. In conclusion, the transformation of secondary metabolites was the dominant trend during HJ growth. These findings lay the foundation for food development and distinguishing clinical applications.

An integrated strategy to reveal the potential anti-asthma mechanism of peimine by metabolite profiling, network pharmacology, and molecular docking.

Peimine, one of the major quality markers in Fritillaria Cirrhosae Bulbus, was expected to become a new anti-asthma drug. However, its metabolic profiles and anti-asthma mechanism have not been clarified previously. In this study, a method was developed for the detection of peimine metabolites in vitro by ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry. The potential anti-asthma mechanism was predicted by an integrated analysis of network pharmacology and molecular docking. A total of 19 metabolites were identified with the aid of software and molecular networking. The metabolic profiles of peimine elucidated that the metabolism was a multi-pathway process with characteristics of species difference. The network pharmacology results showed that peimine and its metabolites could regulate multiple asthma-related targets. The above targets were involved in various regulatory pathways linked to asthma. Moreover, the results of molecular docking showed that both peimine and its metabolites had a certain affinity with the ß2 adrenergic receptor. The results provided not only important references to understand the metabolism and pharmacodynamic changes of peimine in vitro, but also supporting data for further pharmacological evaluation. It also provided a new perspective for clarifying the functional changes of traditional Chinese medicine in vitro.

[Studies on chemical constituents of Helwingia chinensis].

OBJECTIVE: To investigate the chemical constituents of Helwingia Chinensis. METHOD: Compounds were isolated with silica gel, Sephadex LH - 20 and polyamide chromatography, and their structures were elucidated by means of spectral analysis. RESULT: Six compounds were isolated and identified as cinnamic acid (1), gult-5-en-3beta-ol (2), friedelin (3), alpha-amyrin (4), luteolin-7-O-beta-D-glucoside (5) and 4, 5-dimethoxy-1, 2-benzoquinone (6). CONCLUSION: All these Compounds were obtained from H. chinensis for the first time.