Research On Chemical Constituents Of Lu Dangshen And Their Biological Activities

Codonopsis Radix is the dried root of Codonopsis pilosula(Franch.)Nannf.,Codonopsis pilosula Nannf.var.modesta(Nannf.)L.T.Shen or Codonopsis tangshen Oliv.,which has the effects of invigorating the spleen,nourishing the lung and blood.Codonopsis Radix mainly contains chemical components like alkyne glycosides,terpenes,phenylpropanoids,alkaloids and it has many kinds of pharmacological action such as regulating immune function,anti-oxidizing and protecting gastric mucosa.Lu Dangshen is the dried root of Codonopsis pilosula(Franch.)Nannf.It is produced in Guluzhou(now Changzhi and Jincheng)in Shanxi Province.It has a long history of cultivation and also has good quality.It is a famous authentic medicinal material and is widely used as tonic Chinese medicine.It has broad prospects of development and utilization.Predecessors have done more research on the chemical constituents of Codonopsis Radix but less on the chemical constituents of Lu Dangshen.Based on the above situation,clarifying the chemical constituents and pharmacological activitives of Lu Dangshen becomes the focus of the research.We analyzed the chemical constituents of Lu Dangshen on the basis of HPLC-Q-TOF-MS/MS technology;By HPLC-Q-TOF-MS/MS technology,we analyzed the difference of chemical constituents of Codonopsis Radix from different habitats;We filtered and identified the different chemical constituents and found out the changing regularity of their content;Taking Lu Dangshen as the object of research,we separated the 50%ethanol eluting part of Lu Dangshen’s 70%ethanol extract systematically to obtain the monomeric compounds,and conducted activity researches on anti-oxidation and immunity-enhancing.The main content of this research includes the following aspects:1.HPLC-Q-TOF-MS/MS technology was used to analyze the chemical constituents of Lu Dangshen.In this study,32 chemical constituents were identified from Lu Dangshen by combining the accurate mass,secondary high-resolution mass spectroscopy,comparison of reference substances and related literature data.These chemical constituents mainly included nucleosides,amino acids,organic acids,alkaloids,phenylpropanoids,alkyne glycosides,flavonoids and terpenes.2.HPLC-Q-TOF-MS/MS technology was used to compare the differences of the components of Codonopsis Radix from different habitats.Multivariate statistical analysis was used to find out the chemical components and their distributing regularities which were significantly different from others.There were obvious differences in the chemical components of Codonopsis Radix from different habitats.A total of 23 different components were filtered.They were uracil,adenine,guanine,succinic acid,hypoxanthine,salicylic acid,phenylalanine,cinnamic acid,codonopyrrolidium B,tryptophan,vanillic acid,caffeic acid-O-hexoside,syringin,chlorogenic acid,caffeic acid,tangshenoside I,luteolin-7-O-glucopyranoside,rutinum,azelaic acid,lobetyolin,secoisolariciresinol,luteolin and atractylenolide Ⅲ,which presented certain distribution characteristics in Codonopsis Radix from different habitats.3.10 compounds were separated from the 70%ethanol extract of Lu Dangshen using macroporous resin column chromatography,silica gel column chromatography,gel column chromatography and semi-preparative high performance liquid chromatography.Chemical methods,physicochemical properties and spectral data were used to identify the structures of the obtained compounds.The compounds were respectively identified as:taraxerol(1),atractylenolide Ⅲ(2),apigenin(3),(+)-pinoresinol(4),lobetyolin(5),coniferin(6),syringin(7),vanillic acid(8),chlorogenic acid(9)and succinic acid(10),among which compounds 3,4,6,and 9 were firstly separated from Lu Dangshen.4.10 monomeric compounds separated from Lu Dangshen and 10%,30%,50%,70%,95%ethanol eluting parts of the 70%alcohol extract of Lu Dangshen were selected to test there antioxidating ability by DPPH free radical scavenging rate experiment,ABTS+cation free radical scavenging rate experiment and FRAP scavenging rate experiment.The results showed that part of the monomeric compounds and part of the eluting parts separated from Lu Dangshen had certain scavenging effects on DPPH free radicals,ABTS+ free radicals and the free radicals of the FRAP kit,and the scavenging ability increased with the increase of sample’s concentration.5.Network pharmacology were used to study whether the monomeric compounds separated from Lu Dangshen can enhance immunity and its mechanism of action.It was found that among the 10 monomeric compounds,apigenin,taraxerol,vanillic acid and atractylenolide Ⅲ might act on the core targets such as AKT1,MAPK3,SRC and EGFR and participate in platelet activation,peptidyl-tyrosine autophosphorylation,sphosphatidylinositol-mediated signaling,regulation of phosphatidylinositol 3-kinase signaling and leukocyte migration.The 4 monomeric compounds might play the role of enhancing immunity by some pathways,such as proteoglycans in cancer,pathways in cancer,PI3K-Akt signaling pathway,estrogen signaling pathway and focal adhesion.