Primary Comments on Chemotaxonomy of Paris spp. Based on Saponins Analysis

Aim The several species of the genus Paris called ＂Chonglou＂ are famous traditional Chinese herbal medicines. We established the quantitative analysis method of the steroidal saponins in some species of the genus Paris and discussed their relations. Methods We detected the contents of 11 steroidal saponins in Paris samples with a Kromasel C18 （ 150 mm× 4.6 mm ID, 5μm） column which was subjected to gradient elution with acetonitrile-water （30：70- 60：40, V/V） at a flow rate of 1 mL· min^-1 by HPLC-ELSD and established chemical cluster tree using SPSS 11 software. Results All the samples could be separated and calibration curves of 11 saponins were prepared. We successfully detected the contents of 11 steroidal saponins in 14 Paris spp. in 30 min. The recovery for the assay of saponins was between 95 % and 97 %. The RSD of precision of 11 saponins and stability of samples were below 3 %. Chemical phylogenetic tree based on saponin contents indicated that 17 samples of Paris spp. clustered separately. Conclusion The established method is accurate and convenient, and suitable for the quantitative analysis of these 11 steroidal saponins in Paris spp.. The chemical phylogenetic tree is in accordance with Takhtajian classical taxonomy.

Spirostane Steroidal Saponins from the Undergroud Parts of Tupistra chinensis

Two new spirostanol saponins, (25S)-spirostane-1β,3β,5β-triol 3-O-β-D-glucopyrano- side 1 and rhodeasapogenin 3-O-β-D-glucopyranosyl-(1 → 4)-β-D-glucopyranoside 2, together with a known saponin, rhodeasapogenin 3-O-β-D-glucopyranoside 3, were isolated from the underground parts of Tupistra chinensis Bak.. Their structures were determined by spectroscopic analysis.

Furostanol Saponins from Asparagus cochinchinensis and Their Cytotoxicity

Phytochemical investigation on the roots of Asparagus cochinchinensis led to the isolation of one new furostanol saponin,named 26-O-β-d-glucopyranosyl-22α-hydroxyl-(25R)-Δ5(6)-furost-3β,26-diol-3-O-α-l-rhamnopyranosyl-(1→2)-[β-d-glucopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→4)]-β-d-glucopyranoside(1),along with three known congeners(2‒4).The structure of new saponin was elucidated via comprehensive inspection of its HRMS and NMR spectral data as well as chemical technology,whereas those of known ones were identified by comparison of their NMR and MS spectral data with those reported in literatures.All isolated saponins were evaluated for their cytotoxic effects on two human liver(MHCC97H)and lung adenocarcinoma(H1299)cancer cells in vitro.Among them,both 1 and 2 showed significant cytotoxicity against above mentioned cell lines.Further studies revealed that these two saponins could significantly inhibit their proliferation of MHCC97H and H1299 cells.

Steroidal Saponins from the Genus Smilax and Their Biological Activities

The Smilax species,widely distributed in tropical region of the world and the warm areas of East Asia and North America,are extensively used as folk medicine to treat inflammatory disorders.Chemical investigation on Smilax species showed they are rich sources of steroidal saponins with diversified structure types,including spirostane,isospirostane,furostane,pregnane,and cholestane.This review mainly summarizes the steroidal saponins(1–104)reported from the genus Smilax between 1967 and 2016,and their biological activities.The relationship between structures of steroidal saponins and related biological activities were briefly discussed.

Two New Steroidal Saponins from Diuranthera inarticulata

Two new steroidal saponins, diuranthosides D and E, were isolated from the whole plant of Diuranthera inarticulata Wang et K. Y. Lang. By means of spectral and chemical analysis, the structure of the new compounds were established as neotigogenin-3-O-(-D-glucopyranosyl (1?3)-(-D-xylopyranosyl-(1?3)-[(-D-glucopyranosyl(1?2)]-(-D-glucopyranosyl(1?4)-(-D-galactopyranoside (1) and neotigogenin-3-O-(-D-glucopyranosyl(1?3)-(-D-glucopyranosyl (1?3)-(-D-xylopyranosyl(1?3)-[(-D-glucopyranosyl(1?2)]-(-D-glucopyranosyl(1?4)-(-D-galactopyranoside (2) respectively.

Steroidal saponins with anti-inflammatory activity from Tribulus terrestris L.

Objective:Tribulus terrestris L.(T.terrestris)is a highly valuable traditional Chinese medicine used to treat stroke,inflammation,pulmonary fibrosis,liver cancer,and urolithiasis.To identify the basic substance responsible for the anti-inflammatory effect of TST(total saponins of Tribulus),its chemical composition was systematically studied,and its effect of inhibiting nitric oxide generation and the expression of related inflammatory factors were determined.Methods:To separate chemical constituents from T.terrestris by column chromatography.Spectroscopic methods,including 1D and 2D nuclear magnetic resonance spectroscopy(NMR)and mass spectrometry(MS)techniques,were used to elucidate the isolated compounds.The anti-inflammatory activities of TST and several compounds were evaluated in vitro.Results:Fifteen steroidal saponins,including 9 furostanol steroidal saponins(1,2,3,4,5,6,7,8,and 15)and 6 isospirostanol steroidal saponins(9,10,11,12,13,and 14),were isolated from T.terrestris.TST significantly decreased the expression of tumor necrosis factor-a(TNF-a)and interleukin-6(IL-6)in RAW 264.7 cells stimulated by lipopolysaccharides.Compounds 13 and 15 evidently reduced TNF-a expression.Compounds 6,10,12,13,and 15 markedly reduced IL-6 expression.Conclusions:Compounds 1 was a novel furostanol steroidal saponin,named 26-O-b-D-glucopyranosyl-(25R)-5afurostan-12-carbonyl-20(22)-en-3b,26-diol-3-O-{b-D-xylopyranosyl-(1→2)-[b-D-xylopyranosyl-(1→3)]-b-D-glucopyranosyl-(1→4)-[a-L-rhamnopyranosyl-(1→2)]-b-D-galactopyranoside}.Compounds 2 was isolated from the family Zygophyllaceae for the first time,and 5 and 6 were isolated from the Tribulus genus.TST and compounds 6,10,12,13,and 15 exerts antiinflammatory activity.

NEW MINOR STEROIDAL SAPONINS FROM SMILAX LEBRUNII

Two new minor steroidal saponins, (25R) spirostan-3β-ol-6-one-3-O-[α-L- arabinopyranosyl(1-4)]-β-D-glucopyranosidc (1) and (25S) spirost-5-en -3β, 17α, 27-triol-3-O- [α-L-arabinopyranosyl(1-6)]-β-D-glucopyranosidc (2) were isolated from the roots of Smilax lebrunii. Their structures were established on thc basis of chemical and spectral methods.

Two New Steroidal Saponins from Tacca plantaginea

Two new C27 steroidal glycosides, named taccaoside A (1) and B (2), were isolated from the traditional Chinese herb Tacca plantaginea. The spectroscopic and chemical evidences revealed their structures to be 26-O--D-glucopyranosyl-(25R)-3?26-dihydroxy furost-5,20-diene -3-O-[?L-rhamnopyranosyl(12)]-[?L-rhamnopyranosyl(13)]--D-glucopyranoside (1) and 26-O--D-glucopyranosyl-(25R)-3?26-dihydroxy furost-5,20-diene-3-O-[?L-rhamnopyranosyl- (12)]-[?D-glucopyranosyl(13)--L-rhamnopyranosyl(13)]--D-glucopyranoside (2), res- pectively.

Neuroprotection of total steroid saponins from Dioscorea zingiberensis C.H.Wright against transient focal cerebral ischemia-reperfusion injury in rats via its anti-inflammatory and anti-apoptotic effects

OBJECTIVE The total steroid saponins(TSSN)isolated from Dioscorea zingiberensis C.H.Wright(D.zingiberensis)has shown a variety of beneficial bioactivities.However,there are no reports about the neuroprotective effects of the TSSN until now.Therefore,we explored the neuroprotective effects of TSSN on rats against transient focal cerebral ischemia-reperfusion(I/R)and the underlying mechanisms.METHODS The healthy adult Sprague-Dawley rats were randomly assigned into six groups.After pre-treatment with the TSSN intragastrically for six days,the rats were subjected to the ischemia injury by the surgery of middle cerebral artery occlusion(MCAO)for 90 min.Some indexes were evaluated and detected.RESULTS As compared to the I/R group,TSSN group of rats,especially given the 30 mg·kg^-1 of TSSN,not only marked reduction in the neurological deficit scores,cerebral infarct volume,and brain edema,but also an increase in neuron survival(Nissl bodies)in the hippocampal cornuammons 1(CA1)and cortex hemisphere of the ipsilateral ischemia.At the same time,the inflammatory cytokines in serum induced by MCAO were significantly alleviated by the TSSN pre-administration.What′s more,the increase of caspase-3 was evidently reduced in the CA1 and cortex of the hemisphere injured brain.Finally,the down-regulating anti-apoptotic Bcl-2 and up-regulating pro-apoptotic Bax proteins were obviously suppressed.CONCLUSION TSSN plays a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model,and this role may be mediated by its antiinflammatory and anti-apoptotic actions.

Untiring Researches for Alternative Resources of Rhizoma Paridis

Rhizoma Paridis(RP,M®),a traditional Chinese medicine,is the rhizoma of Paris polyphylla var.yunnanensis(PPY)or P.polyphylla var.chinensis which are widely used as important raw materials for several Chinese patent drugs.However,the wild resources of these herbs have become less and less due to their slow-growing characteristics and previously excessive excavation.This review covers untiring investigations on alternative resources of RP by our research group over the past decades,including non-medicinal parts of PPY as well as other plants of Liliaceae and Liliflorae families.The arial parts of PPY and the whole plants of Trillium kamtschaticum might be alternative resources for RP based on the fact that they shared the same or similar saponins and bioactivities.

The Genus Solanum:An Ethnopharmacological,Phytochemical and Biological Properties Review

Over the past 30 years,the genus Solanum has received considerable attention in chemical and biological studies.Solanum is the largest genus in the family Solanaceae,comprising of about 2000 species distributed in the subtropical and tropical regions of Africa,Australia,and parts of Asia,e.g.,China,India and Japan.Many of them are economically signifcant species.Previous phytochemical investigations on Solanum species led to the identifcation of steroidal saponins,steroidal alkaloids,terpenes,favonoids,lignans,sterols,phenolic comopunds,coumarins,amongst other compounds.Many species belonging to this genus present huge range of pharmacological activities such as cytotoxicity to diferent tumors as breast cancer(4T1 and EMT),colorectal cancer(HCT116,HT29,and SW480),and prostate cancer(DU145)cell lines.The biological activities have been attributed to a number of steroidal saponins,steroidal alkaloids and phenols.This review features 65 phytochemically studied species of Solanum between 1990 and 2018,fetched from SciFinder,Pubmed,ScienceDirect,Wikipedia and Baidu,using“Solanum”and the species’names as search terms(“all felds”).

A New Furostanol Saponin from the Water-extract of Dioscorea nipponica Mak.,the Raw Material of the Traditional Chinese Herbal Medicine Wei Ao Xin

O-a-D-Glucopyranosyl-furost-5(6),20(22)-dien-3a,26-diol 1, a new furostanol saponin, was isolated from the water-extract of Dioscorea nipponica Mak., the raw material of a traditional Chinese herbal medicine Wei Ao Xin. The structure of 1 was determined on the basis of its spectral data especially by NMR spectroscopy. The result provides the first example of naturally occurring furostanol saponins with a single saccharide chain at the C-26 position.

Phytochemical and pharmacological studies on Solanum lyratum:a review

Solanum lyratum is one of the temperate plants,broadly distributed in Korea,China,Japan,India,and South-East Asia and well-documented in those oriental ethnic medicine systems for curing cancers,jaundice,edema,gonorrhea,cholecystitis,phlogosis,rheumatoid arthritis,etc.This review systematically summarized the research progress on S.lyratum respecting the botany,traditional uses,phytochemistry,pharmacology,and toxicology to increase people’s in-depth understanding of this plant,by data retrieval in a series of online or off-line electronic databases as far as we can reach.Steroidal saponins and alkaloids,terpenoids,nitrogenous compounds,and flavonoid compounds are the main chemical constituents in S.lyratum.Among them,steroidal alkaloids and saponins are the major active ingredi-ents ever found in S.lyratum,exerting activities of anti-cancer,anti-inflammation,anti-microbial,anti-allergy,and anti-oxidation in vivo or in vitro.As a result,S.lyratum has been frequently prescribed for the abovementioned therapeutic purposes,and there are substantial traditional and modern shreds of evidence of its use.

Unraveling the serial glycosylation in the biosynthesis of steroidal saponins in the medicinal plant Paris polyphylla and their antifungal action

Sugar-sugar glycosyltransferases play important roles in constructing complex and bioactive saponins.Here,we characterized a series of UDP-glycosyltransferases responsible for biosynthesizing the branched sugar chain of bioactive steroidal saponins from a widely known medicinal plant Paris polyphylla var.yunnanensis.Among them,a 2'-O-rhamnosyltransferase and three 6'-O-glucosyltrasferases catalyzed a cascade of glycosylation to produce steroidal diglycosides and triglycosides,respectively.These UDP-glycosyltransferases showed astonishing substrate promiscuity,resulting in the generation of a panel of 24 terpenoid glycosides including 15 previously undescribed compounds.A mutant library containing 44 variants was constructed based on the identification of critical residues by molecular docking simulations and protein model alignments,and a mutant UGT91AH1^(Y187A)with increased catalytic efficiency was obtained.The steroidal saponins exhibited remarkable antifungal activity against four widespread strains of human pathogenic fungi attributed to ergosterol-dependent damage of fungal cell membranes,and 2'-O-rhamnosylation appeared to correlate with strong antifungal effects.The findings elucidated the biosynthetic machinery for their production of steroidal saponins and revealed their potential as new antifungal agents.

Limonoid and Steroidal Saponin from Azadirachta indica

A new limonoid,17-(5-methoxy-2-oxofuran-3-yl)-28-deoxonimbolide(1),and a new C21 steroidal saponin,2a,4a-dihydroxy-pregn-5-en-16-one-3a-O-D-glucopyranoside(2),together with 11 known compounds were isolated from the methanol extract of the leaves of Azadirachta indica.The structures were elucidated by means of spectroscopic analysis and putative biosynthetic origins.All the compounds were evaluated for their antibacterial activities against six bacterial strains.

A NEW STEROIDAL SAPONIN FROM SMILAX LEBRUNH

A New steroidal saponin was isolated from the rhizomes of Smilax lebrunii. On the basis of spectral and chemical methods, its structure was elucidated as:(25S)spirost-5-en-3β, 17α, 27-triol-3-0-[β-D-glucopyranosyl(1-4)][α-L-arabinopyranosyl(1-6)]-β-D-glucopyranoside.

Steroid and triterpenoid saponins from the rhizomes of Paris polyphylla var.stenophylla

Five new saponins,including three steroid saponins,paristenoids A-C(1-3),and two triterpenoid saponins,paristenoids D-E(4-5),along with four known ones(6-9)were isolated from the rhizomes of Paris polyphylla var.stenophylla.The structures of the isolated compounds were identified nainly by detailed spectroscopic analysis,including extensive 1D and 2D NMR,MS,as well as chemical methods.Compound 3 is a new cyclocholestanol-type steroidal saponin with a rare 6/6/6/5/5 fused-rings cholestanol skeleton,and this skeleton has been first found from the genus Paris.The cytotoxicities of the isolated compounds against three human three glioma cell lines(U87MG,U251MG and SHG44)were evaluated,and compound 7 displayed certain inhibitory effect with ICso values of 15.22±1.73,18.87±1.81 and 17.64±1.69μmol^(-1),respectively.

Simultaneous determination of steroidal saponins in Anemarrhena asphodeloides Bge. by ultra high-performance liquid chromatography/ quadrupole time-of-flight mass spectrometry

The objective of the research is to develop a quantitative method by ultra high-performance liquid chromatography/ quadrupole time-of-flight mass spectrometry （UHPLC/Q-TOF MS） for the analysis of seven major steroidal saponins （timosaponin N, timosaponin El, timosaponin BII, timosaponin B, anemarrhenasaponin I, anemarrhenasaponin A2, and timosaponin AIII） in Anemarrhena asphodeloides Bge. The complete separation of these seven steroidal saponins was achieved within 18 min with an ACQUITY UPLC HSS T3 column using an acetonitrile-water （contain 0.1% formic acid） gradient system. The limits of quantitation （LOQ）, 0.18-0.75 ng/pL for seven steroidal saponins, were determined experimentally. The limits of detection （LOD） were found to be 0.05-0.22 ng/μL for these saponins. The correlation coefficients （r2） for calibration curves varied from 0.9902 to 0.9979. This method showed good repeatability for the quantification of these saponins in rhizomes ofA. asphodeloides, with intra-day and inter-day variations of less than 5.0% for seven steroidal saponins. The recoveries of seven steroidal saponins were from 97.13% to 101.98%. The validated method was successfully applied to quantifying seven steroidal saponins in various sources ofA. asphodeloides （different collecting places or processing methods） and Zhimu concentrate-granules （ZMCG）.

Arbuscular mycorrhiza and plant growth promoting endophytes facilitates accumulation of saponin under moderate drought stress

Objective: Paris polyphylla var. yunnanensis, one of the important medicinal plant resources in Yunnan,China, usually takes 6–8 years to be harvested. Therefore, it is urgent to find a method that can not only shorten its growth years, but also improve its quality. In this study, we examined the effects of a combination treatment of arbuscular mycorrhizal fungi(AMF) and plant growth-promoting endophytes(PGPE)and drought stress on the accumulation of saponins in it.Methods: P. polyphylla var. yunnanensis was infected with a mixture of AMF and PGPE under drought stress. The content of saponins, as well as morphological, physiological, and biochemical indicators, were all measured. The UGTs gene related to saponin synthesis was obtained from transcriptome data by homologous comparison, which were used for RT-PCR and phylogenetic analysis.Results: Regardless of water, AMF treatment could infect the roots of P. polyphylla var. yunnanensis, however double inoculation with AMF and PGPE(AMF + PGPE) would reduce the infection rate of AMF. Plant height, aboveground and underground fresh weight did not differ significantly between the single inoculation AMF and the double inoculation treatment under different water conditions, but the inoculation treatment significantly increased the plant height of P. polyphylla var. yunnanensis compared to the noninoculation treatment. Single inoculation with AMF considerably increased the net photosynthetic rate,stomatal conductance, and transpiration rate of P. polyphylla var. yunnanensis leaves under various water conditions, but double inoculation with AMF + PGPE greatly increased the intercellular CO2concentration and chlorophyll fluorescence parameter(Fv/Fm). Under diverse water treatments, single inoculation AMF had the highest proline content, whereas double inoculation AMF + PGPE may greatly improve the amount of abscisic acid(ABA) and indoleacetic acid(IAA) compared to normal water under moderate drought. Double inoculation AMF + PGPE treatment improved the proportion of N, P, and K in the rhizome of P. polyphylla var. yunnanensis under various water conditions. Under moderate drought stress,AMF + PGPE significantly enhanced the contents of P. polyphylla var. yunnanensis saponins I, II, VII, and total saponins as compared to normal water circumstances. Farnesyl diphosphate synthase(FPPS),Geranyl pyrophosphate synthase(GPPS), Cycloartenol synthase(CAS), and Squalene epoxidase(SE1) were the genes that were significantly up-regulated at the same time. The amount of saponins was favorably linked with the expression of CAS, GPPS, and SE1. Saponin VI content and glycosyl transferase(UGT)010922 gene expression were found to be substantially associated, as was saponin II content and UGT010935 gene expression.Conclusion: Under moderate drought, AMF + PGPE was more conducive to the increase of hormone content, nutrient absorption, and total saponin content in P. polyphylla var. yunnanensis, and AMF + PGPE could up regulate the expression of key genes and UGTs genes in one or more steroidal saponin synthesis pathways to varying degrees, thereby stimulating the synthesis and accumulation of steroidal saponins in the rhizome of P. polyphylla var. yunnanensis. The combination of AMF and PGPE inoculation, as well as adequate soil drought, reduced the buildup of saponins in P. polyphylla var. yunnanensis and increased its quality.

Fingerprint analysis of Zhimu-Huangbai herb pair and simultaneous determination of its alkaloids, xanthone glycosides and steroidal saponins by HPLC-DAD-ELSD

AIM: To develop and validate a high performance liquid chromatography(HPLC) coupled with diode array and evaporative light scattering detectors(DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes(namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair(ZB). METHOD: Chromatographic separation was performed on a Diamonsil C18 column(4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min–1 at 260 nm. The drift tube temperature of ELSD was set to 60 ℃ and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS: The HPLC-DAD-ELSD method allowed the quantification of ten compounds(phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION: This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.