| Medicinal plants are the main source of traditional medicines in all nationalities all over the world for modern drug development and utilization.The collation,excavation,and modern research of important medicinal plants have been an important part of the inheritance and innovative development of Chinese medicine.Plants of the genus Scutellaria in the family Lamiaceae are widely distributed worldwide,with about 360-469 species,of which about 72 species or varieties are used for medicinal purposes.There are about 126 species or varieties in China,of which about 45 species or varieties have been recorded for traditional medicinal use.Modern studies have shown that this genus has various activities,such as antitumor,anti-inflammatory,antioxidant,antiviral.hepatoprotective,and neuroprotective.It is showing that Scutellaria contains rich medicinal plants,which worthy of further exploration and utilization.However,except for a few species,such as Scutellaria baicalensis Georgi and Scutellaria barbata D.Don,the chemical composition and pharmacological activity of most plants in the genus Scutellaria have been little or no research.In addition,due to a large number of species and complex trait variation,the taxonomy of this genus is still unclear.Thus,a comprehensive study of this genus from a systems biology perspective is needed to reveal the pharmacophylogenetic of Scutellaria,to discover new medicinal resources and active ingredients.In this paper,based on the theory of pharmacophylogeny,the chloroplast genomes used to reveal the phylogeny of Scutellaria;metabolomic techniques and chemical composition analysis to reveal the distribution patterns of the major components of the genus;in vivolin vivo activity studies to reveal the antitumor activity characteristics of the genus.Besides,systematic analysis of the phylogeny-chemical constituents-pharmaceutical effects relationship of Scutellaria was conducted.In addition,the possible internal relations among phylogeny,chemical composition,and activity of Scutellaria were systematically analyzed.The details of the study and the results are as follows:1.Due to the large number of species and the complexity of trait variation in Scutellaria,the classical morphological classification is highly divergent and has not been effectively unified.In this study,a phylogenetic tree was constructed using the chloroplast genomes to provide a reference background for rational exploration of the pharmacophylogenetic of Scutellaria.Th e full length of 24 species of Scutellaria chloroplast genomes were determined to be 151 574-152 591 bp with an average GC content of 38.34%.A total of 113 genes were annotated,including 80 protein-coding genes,29 tRNAs,and four rRNAs.The comparative analysis with the published chloroplast genomes of Scutellaria and phylogenetic tree construction was also performed.Through comparative genomic analysis,abundant simple sequence repeat loci were detected in these species,among which single nucleotide repeats accounted for the highest proportion and pentanucleotide repeats accounted for the lowest proportion.In the chloroplast genomes of these species of Scutellaria,16 hypervariable regions are proposed as a potential DNA barcode.The results of the study not only revealed the preliminary relatives of the major medicinal species of Scutellaria in China but also promoted the construction of the molecular system of Scutellaria.2.Natural products are an important source of active substance base and new drug discovery for medicinal plants.The species of Scutellaria have important medicinal values,but most of the chemical components in this genus are shallowly studied or still blank.Systematic analysis and comparison of chemical composition distribution patterns of this genus is the basis for revealing the pharmacophylogeny of Scutellaria and discovering potential medicinal resources and active ingredients.In this study,the metabolite compositions of 35 species of Scutellaria in China were performed by UPLC-Q-TOF-MS for the first time.A total of 196 compounds were identified or annotated in this study,including 118 flavonoids,30 diterpenoids,5 pyranosides,3 phenylethanoid glycosides,and 39 other compounds.Among them,85 compounds were annotated in Scutellaria for the first time.Principal component analysis based on the characteristic data matrix grouped the 35 species into basically two groups,which supported to some extent the phylogenetic tree of Scutellaria constructed based on the chloroplast genomes.In addition,the differential chemical constituents accumulated by different groups of Scutellaria based on chemical composition clustering were identified separately.It was found that the characteristic chemical constituents contained in different taxa of Scutellaria may have some association with their respective traditional efficacy.However,the limited data available at present do not allow us to draw a clear relationship between the differential chemical constituents and their therapeutic efficacy.The results of the study basically reveal the distribution pattern of chemical components of Scutellaria and provide basic data for the elucidation of pharmacophylogeny of Scutellaria,and also lay the foundation for the development and utilization of potential medicinal plant resources of Scutellaria.3.Some of the medicinal plants and compounds in the genus of Scutellaria displayed significant antitumor activities.However,the global analysis of the antitumor effects and molecular mechanisms of Scutellaria plants is still limited.In this study,the molecular network of antitumor active ingredient-target-pathway-disease was established by the network pharmacology method for the first time.The results showed that:flavonoids and diterpenoids were the main active compounds of the antitumor activity of Scutellaria.Among them,the top 10 potential key active compounds including apigenin,baicalein,and scutebarbatine G.Besides,86 related targets and 203 tumor-related pathways were screened,and key targets obtained by topological screening are AKT1,EGFR,VEGFA and HSP90AA1,etc.;GO functional and KEGG enrichment analysis showed that Scutellaria may inhibit tumor cell proliferation,invasion,inhibit tumor angiogenesis,and reduce inflammatory response through regulating signaling pathways,such as PI3K-Ak,Estrogen,HIF-1,and FoxO.The results of molecular docking showed that the potential compounds had a stable binding activity with the core targets.To evaluate the results of the network pharmacological study,14 compounds,including apigenin,baicalin,acacetin,and 35 methanolic extracts of Scutellaria plants were further determined based on three human-Derived tumor cells using an in vitro tumor cell growth inhibition method.The metabolomics and content of major active compounds were determined for the analysis of the more active extract.The results showed that the free flavonoid generally showed better inhibitory activities against various tumor cells,especially baicalin and luteolin showed significant inhibitory activities against human cancers;In addition,it was found that S.pekinensis var.ussuriensis exhibited more prominent inhibition of human hepatocellular cancer Bel7402 cells and human colorectal cancer HCT116 cells,and S.pekinensis showed better inhibition of human colorectal cancer HCT116 cells.The compounds of chrysin,luteolin,apigenin,baicalin,and wogonin may be the main active compounds exerting antitumor activity of these two species.Besides,the roots of S.amoena also showed well inhibitory effect on these three tumor cells.Baicalein,chrysin,wogonin as well as high contents of baicalin,wogonoside,scutellarin,and oroxylin Amay be its main active compounds.The results not only partially validated the results of the network pharmacology analysis but also provided a direction for further research on the antitumor activity of Scutellaria.In addition,more than half of the extracts of Scutellaria species in this experiment were studied for the first time for their in vitro antitumor activity,which can provide a useful reference for the discovery of antitumor active resources of Scutellaria.4.Huang-qin tea is prepared from the aerial parts of S.baicalensis,which is traditionally used as a tea substitute with the effects of clearing heat,drying dampness,reducing fire,and detoxify,promoting digestion.Based on the analysis of pharmacophylogeny of Scutellaria and previous studies,it was suggested that Huang-qin tea might have better anti-tumor effects.To verify this hypothesis,this study investigated the chemopreventive effect of Huang-qin tea against colorectal cancer and its mechanism.The results showed that Huang-qin tea significantly inhibits azoxymethane(AOM)-induced preneoplastic colonic aberrant crypt foci(ACF)formation.Besides,Huang-qin tea modulates inflammatory cytokines expression by significantly decreasing IL-1?,IL-6,IL-10.TNF-? expression,and elevating IFN-y production.The essential role of gut microbiota was confirmed for Huang-qin tea to prevent CRC by the pseudo-germ-free rat.16S rDNA sequencing analysis indicated that Huang-qin tea regulated gut microbiota by increasing the abundance of beneficial bacteria(Lachnoclostridium,Alistipes,Roseburia.Lactococcus)and reducing the level of Bacteroides,Parasutterella,and unidentified_Clostridiales.Fecal metabolomics showed that Huang-qin tea modulated the AOM-induced metabolomic disorder,and these altered metabolites were almost involved in lipid metabolism pathways.Spearman correlation analysis reveals a correlation between gut microbiota and fecal metabolites.Collectively,these results suggested that Huang-qin tea exerted beneficial effects on the host health by inhibiting inflammation,regulating gut microbiota profile,and certain metabolism pathways.In conclusion,Huang-qin tea inhibits AOM-induced ACF formation by modulating gut microbiota composition and improving metabolomic disorders,indicating the potential of Huang-qin tea as a functional beverage candidate to prevent and treat CRC.5.To reveal the pharmacophylogeny of Scutellarin,this paper explores the relationship between phylogeny,chemical constituents,and pharmaceutical effects of Scutellaria in China based on the above findings and relevant literature reports.Plants of the Subg.Anaspis,Sect.Lupulinaria and Subsect.Angustifoliae are closely related and clustered into one taxon,all with well-Developed roots.The roots of plants in this taxon are often used as herbal medicine.These roots contain high levels of flavonoids that may be associated with the traditional therapeutic effects of clearing heat and removing dampness and the pharmacological activities of antitumor,anti-inflammatory,antioxidant,and hepatoprotective.In addition,plants of the Subgen.Scutellariopsis and other taxa of the subgenus Scutellaria can be grouped into a large category.Plants in this broad group usually have undeveloped roots and the whole herbs are often used as herbal medicines.Flavonoids and/or diterpenoids are the main compounds in this group of plants,which may be associated with traditional effects such as clearing heat,detoxifying,dispelling dampness,reducing carbuncles and swelling,and pharmacological activities of antitumor and anti-inflammatory.The exploration of the pharmacophylogeny of Scutellaria in China can provide a scientific and theoretical basis for the in-Depth development of the medicinal plant resources of Scutellaria.In summary,this paper integrates multi-level research data from the perspectives of molecular phylogeny,metabolomics,and antitumor pharmacological activity to investigate the pharmacophylogeny of Scutellaria in China.The results of the study can provide directions for the discovery of active natural products and antitumor active plant resources of Scutellaria,and lay a good foundation for further in-Depth research and exploitation of this genus. |
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