| Purpose:Fallopia denticulata(C.C.Huang)T.M.Schust.& Reveal,a dried tuberous root of Fallopia denticulata,family Polygonaceae,has been used to treat indigestion,stomach pain and influenza.However,the existing taxonomic studies on the original plant,P.denticulatum,are confusing and there is no unified classification standard,and there are few articles on the classification of P.denticulatum,which causes the basal origin of the plant to be unknown;the plant is commonly used as an anti-influenza medicine by the Yi folk,but no scholars have investigated the anti-influenza activity of the plant;the plant is widely distributed in Yunnan and Guizhou,but there are few studies on its chemical composition and biological activity.This has also resulted in the waste of the resources of P.denticulatum.Methods:(1)We firstly selected the chloroplast genomes of P.denticulatum,Fallopia multiflora,Pleuropterus ciliinrvis,Fallopia aubertii,Fallopia dentatoalata,and Fallopia convolvulus as the subjects of this paper,and obtained the chloroplast genomes of P.denticulatum,Pleuropterus ciliinrvis,Fallopia dentatoalata,and Fallopia convolvulus for the first time.The chloroplast genomes of P.denticulatum,F.multiflora,P.ciliinrvis,F.aubertii,F.dentatoalata,and F.convolvulus were compared in terms of genome composition,structure,codon preference,and repetitive sequences,etc.The phylogeny of P.denticulatum was analyzed by chloroplast genomics,and specific barcodes were designed and validated to identify the six species,reconstructing the phylogenetic relationships of P.denticulatum and clarifying the genesis of P.denticulatum.(2)The extraction process of anti-neuraminidase active site of P.denticulatum was established,the extraction solvents with different polarity were analyzed by neuraminidase inhibition experiments,and the extracts with the best extraction solvents were later analyzed by UPLC-Q-TOF-MS technique.(3)This paper conducted a broad-target metabolomics analysis of P.denticulatum,and a systematic investigation of the compounds of P.denticulatum,combining the secondary metabolites from the downscaled analysis with the network pharmacology and molecular docking virtual screening techniques,providing a theoretical basis for the antidiabetic effect of P.denticulatum and providing literature support for the utilization of P.denticulatum resources.Results:(1)By reviewing the literature,the research progress of the classification of the original plant P.denticulatum,chemical constituents and pharmacological effects of P.denticulatum at home and abroad was reviewed;at the same time,the latest research on chloroplast genomics and chloroplast genome of Polygonaceae was systematically summarized.The progress and the latest research status of metabolomics provide a theoretical basis for determining the perfect research content and formulating a feasible research design for this paper.(2)The results of phylogenetic analysis of P.denticulatum show that:(i)the chloroplast whole genomes of P.denticulatum,F.multiflora,P.ciliinrvis,F.aubertii,F.dentatoalata,and F.convolvulus were similar in structure,genome size,gene composition and genome sequence,and the gene distribution position and gene length at the boundary between LSC,SSC and IR were almost identical,with different degrees of expansion and contraction,showing that rps19 and ndh F genes were lost in the IRa region.The topology of phylogenetic trees constructed by Maximum Likelihood(ML),Maximum Parsimony(MP)and Neighbor-Joining(NJ)based on chloroplast coding sequence(CDS)data and chloroplast whole genome data The results support the classification of P.denticulatum in the genus Pteroxygonum in the Encyclopedia of Multicultural Plants,and suggest that F.multiflora,P.ciliinrvis,F.aubertii,F.dentatoalata,and F.convolvulus are sister groups to each other.Four DNA markers with intervals pet N-psb M,psa J-rpl33,ycf3-trn S-GGA and trn L-UAG-ccs A were developed and used for the identification of the six species.(3)The research on the anti-influenza activity of P.denticulatum shows that the extraction process of anti-neuraminidase active parts of P.denticulatum is established,and it is clear that the anti-neuraminidase component of P.denticulatum contains 21 kinds of compounds,namely flavonoids,phenolic acids,alkaloids and others.kind.Among them,(-)-epicatechin gallate,(+)-catechin and(-)-epigallocatechin compounds have strong neuraminidase activity,and have relatively higher anti-influenza activity.great potential.(4)The research results of the anti-diabetic activity of P.denticulatum show that:(1)Based on extensive targeted metabolomics,it was initially determined that P.denticulatum contained 881 compounds,of which 419 were secondary metabolites and462 were primary metabolites,and phenolic acids and flavonoids were the main secondary metabolites of P.denticulatum,with 183 and 83 compounds,respectively.(ii)The network pharmacological analysis showed that(2)Network pharmacological analysis showed that tumor necrosis factor(TNF),Toll-like receptor 4(TLR4),Rela proto-oncogene(RELA),etc.were the core targets of P.denticulatum against diabetes.It is possible that P.denticulatum may treat diabetes by modulating the inflammatory mediators of TRP channel,AGE-RAGE signaling pathway of diabetic complications and prolactin signaling pathway.The results of compound composition-target of actionpathway network analysis showed that the blood entry components with large degree values include Dibutyl phthalate,Isoferulic Acid and N-Feruloylputrescine.(iii)Molecular docking studies showed that 19 compounds showed strong antidiabetic activity,among which Cycloartenol,Alphitolic acid and 2α,19α-Dihydroxy-3-oxours-12-en-28-oic acid compounds deserve further investigation.Conclusion:(1)This study was the first to determine the whole chloroplast genomes of P.denticulatum,P.ciliinrvis,F.dentatoalata,and F.convolvulus,and re-sequenced the whole chloroplast genomes of F.multiflora and F.aubertii,and assembled and annotated six chloroplast genomes.We reconstructed the phylogeny of Polygonum serrata,and developed DNA barcode fragments to identify six species;(2)Based on the phylogenetic analysis of the chloroplast genome,this paper proposes to assign the original plant Polygonum serrata to the genus Fallopia;(3)The anti-influenza activity of P.denticulatum was investigated for the first time,and the extraction process of anti-influenza activity of P.denticulatum was established,and the anti-influenza active components of P.denticulatum were clarified as(-)-epigallocatechin gallate,(+)-catechin and(-)-epigallocatechin;(4)For the first time,the chemical composition of the sauce was systematically explored,and it was clarified that the chemical composition of the sauce contained 881 compounds,including tumor necrosis factor(TNF),Toll-like receptor 4(TLR4),Rela proto-oncogene(RELA),etc.The core target of anti-diabetes.P.denticulatum may treat diabetes by regulating inflammatory mediators of TRP channels,AGE-RAGE signaling pathway and prolactin signaling pathway in diabetic complications.The 19 compounds of P.denticulatum showed strong antidiabetic activity,among which Cycloartenol,Alphitolic acid and 2α,19α-Dihydroxy-3-oxours-12-en-28-oic acid compounds deserve further exploration. |
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