Gut microbiota-mediated metabolism of Panax notoginseng saponins and its role in pharmacokinetics and pharmacodynamics

Panax notoginseng saponins(PNS)are a class of effective ingredients in Notoginseng Radix et Rhizoma,a well-known herbal medicine called San-Qi in Chinese.After oral administration,PNS inevitably interacts with gut microbiota,and thus affect the pharmacokinetic profiles and pharmacological effects.To date,studies concering gut microbiota-mediated metabolism of PNS have not been reviewed systematically.Herein,we outline the metabolic profiles of Panax notoginseng saponins mediated by gut microbiota,as well as its role in the pharmacokinetics and pharmacodynamics on the basis of reported data.The metabolic pathways of primary saponins are proposed,and step-by-step deglycosylation is found to be the primary degradation pathways of PNS mediated by gut microbiota.Specific microorganisms and enzymes involved in the metabolic processes were summarized.Gut microbiota is deeply involved in the metabolism of PNS,affects the pharmacokinetic profiles,and produces a series of active metabolites.These metabolites were documented to play an essential role in the efficacy of the parent compounds.Future studies should focus on strengthening the real-world evidence,defining the interaction between gut microbiota and PNS,and developing the strategy for modulating gut microbiota to enhance the bioavailability and efficacy of PNS.These information would be useful for further research and clinical application of PNS.

Antioxidant,antimicrobial,and α-glucosidase inhibitory activities of saponin extracts from walnut(Juglans regia L.) leaves

Objective:To investigate the relationship between triterpenoid saponin content and antioxidant,antimicrobial,and α-glucosidase inhibitory activities of 70%ethanolic,butanolic,aqueous,supernate and precipitate extracts of Juglans regia leaves.Methods:Triterpenoid saponins of different Juglans regia leaf extracts were measured by the vanillin method.Antioxidant activity was evaluated against DPPH and ABTS free radicals.We also assessed α-glucosidase inhibitory and antimicrobial activities of the leaf extracts.Pearson’s correlation coefficient was evaluated to determine the correlation between the saponin content and biological activities.Results:The butanolic extract was most effective against DPPH with an IC50of 6.63μg/mL,while the aqueous extract showed the highest scavenging activity against ABTS free radical with an IC50of 42.27μg/mL.Pearson’s correlation analysis indicated a strong negative correlation (r=-0.956) between DPPH radical scavenging activity (IC50) and the saponin content in the samples examined.In addition,the aqueous extract showed the best α-glucosidase inhibitory activity compared with other extracts.All the extracts had fair antibacterial activity against Bacillus subtilis,Escherichia coli,and Klebsiella pneumoniae except for the aqueous extract.Conclusions:Juglans regia extracts show potent antioxidant,antimicrobial,and α-glucosidase inhibitory activities.There is a correlation between saponin levels in Juglans regia leaf extracts and the studied activities.However,additional research is required to establish these relationships by identifying the specific saponin molecules responsible for these activities and elucidating their mechanisms of action.

PnMYB4 negatively modulates saponin biosynthesis in Panax notoginseng through interplay with PnMYB1

Saponins are the main triterpenoid ingredients from Panax notoginseng,a well-known Chinese medicine,and are important sources for producing drugs to prevent and treat cerebrovascular and cardiovascular diseases.However,the transcriptional regulatory network of saponin biosynthesis in P.notoginseng is largely unknown.In the present study we demonstrated that one R2R3-MYB transcription factor,designated PnMYB4,acts as a repressor of saponin accumulation.Suppression of PnMYB4 in P.notoginseng calli significantly increased the saponin content and the expression level of saponin biosynthetic genes.PnMYB4 directly bound to the promoters of key saponin biosynthetic genes,including PnSS,PnSE,and PnDS,to repress saponin accumulation.PnMYB4 and the activator PnMYB1 could inter-acted with PnbHLH,which is a positive regulator of saponin biosynthesis,to modulate the biosynthesis of saponin.PnMYB4 competed with PnMYB1 for binding to PnbHLH,repressing activation of the promoters of saponin structural genes induced by the PnMYB1-PnbHLH complex.Our study reveals that a complex regulatory module of saponin biosynthesis is associated with positive and negative MYB transcriptional regulators and provides a theoretical basis for improving the content of saponins and efficacy of P.notoginseng.

Rhizoma paridis saponins protected against liver injury in diethylnitrosamine-induced mice

Background:Diethylnitrosamine,one of food additives,possessed a strong carcinogenic effect in human.Rhizoma paridis saponins,as the main active components of Paris polyphylla,have a good anti-cancer effect in our previous research.To verify their inhibitory effect on diethylnitrosamine-induced liver cancer,we carried out this study.Methods:We established diethylnitrosamine-induced mouse hepatocarcinoma models to evaluate antitumor of Rhizoma paridis saponins.Subsequently,gas chromatography-mass spectrometry was applied to analyze the metabolites in the urine and serum samples.Results:Rhizoma paridis saponins alleviated diethylnitrosamine-induced hepatocarcinogenesis.On the one hand,Rhizoma paridis saponins down-regulated the levels of liver function markers,such as alanine aminotransferase,aspartate transaminase and alpha fetoprotein.On the other hand,Rhizoma paridis saponins reduced metabolic disorders by increasing fructose and mannose metabolism,and decreasing pentose and glucuronate interconversion,inositol phosphate metabolism,and the process of saturated fatty acids transforming to unsaturated fatty acids,which based on the regulating mRNA expression of glucose transporter type 4,lactate dehydrogenase A,fatty acid synthetas,acetyl-CoA carboxylase and apolipoprotein A-I.Conclusion:Rhizoma paridis saponins has the potential application to inhibit chemical-induced hepatocarcinogenesis in the future.

Effects of Tribulus terrestris saponins on proliferation and invasion of A549 cells

Objective:Tribulus terrestris saponin is a traditional Chinese medicine in China.This experiment was designed to investigate the effects of tribulus terrestris saponin on the proliferation and invasion ability of non-small cell lung cancer A549 cells.Methods:A549 cells were divided into normal control and experimental groups(Tribulus terrestris saponin 250μg/mL group,Tribulus terrestris saponin 200μg/mL group,Tribulus terrestris saponin 150μg/mL group,Tribulus terrestris saponin 100μg/mL group,Tribulus terrestris saponin 50μg/mL group).The proliferation viability of the cells in each group was detected by CCK8,the invasion of tumor cells was detected by Transwell model.The mRNA expression of MMP9 and caspase-3 in each group of cells was detected by RT-PCR.Immunofluorescence staining was used to observe the fluorescence intensity of caspase-3 in each group of cells.Results:Compared with the normal control group,tribulus terrestris saponin significantly inhibited the proliferation activity and invasion ability of A549 cells,which was statistically significant(P<0.01).In the invasion assay,compared with the control group,MMP9 expression was significantly reduced and caspase-3 expression was significantly increased in the tribulus terrestris saponin group,and both were concentration-dependent,with statistically significant differences(P<0.01).By cellular immunofluorescence staining experiments,it was found that the fluorescence expression of caspase-3 was enhanced in the experimental group compared with the normal control group,in which the high concentration saponin group was significantly higher than the low concentration group.Conclusion:Tribulus terrestris saponin can inhibit the invasive ability of A549 cells by down-regulating the expression of MMP9,and induce irreversible apoptosis by up-regulating the activation of caspase-3 expression to form caspase-3.

Effects of Arbuscular Mycorrhizal Fungi on the Physiology and Saponin Synthesis of Paris polyphylla var. yunnanensis at Different Nitrogen Levels

Arbuscular mycorrhizal fungi(AMF)are important members of the plant microbiome and affect the uptake and transfer of mineral elements by forming a symbiotic relationship with plant roots.Nitrogen(N),as an important min­eral element,can directly affect plant growth and development at different N levels.It has been confirmed that inoc­ulation with AMF can improve the efficiency of N utilization by plants.However,there are still fewer reports on the dynamic relationship between arbuscular mycorrhizal and plant secondary metabolites at different nitrogen levels.In this experiment,the physiological indexes and genes related to saponin synthesis were determined by applying differ­ent concentration gradients of nitrogen to the medicinal plant P.polyphylla var.yunnanensis infested by AMF as the test material.It was found that nitrogen addition increased the biomass,chlorophyll content,and nutrient content of above-and below-ground plant parts and increased the content of saponin content of P.polyphylla var.yunnanensis to some extent,but AMF inoculation increased the saponin content of P.polyphylla var.yunnanensis more significantly.AMF inoculation also promoted the expression of genes related to the saponin synthesis pathway,including 3-hy­droxy-3-methylglutaryl coenzyme A synthase(HMGS),squalene epoxidase 1(SE1),and cycloartenol synthase(CAS),which is in according with the accumulation of saponin in plants.It also may increase the saponin content of AMF plants by altering the expression of P450s and UGTs related to saponin synthesis.

Two new saponins from Anemarrhena asphodeloides Bge.

Two new saponins 3-O-β-d-glucopyranosyl (1 → 2)-β-d-mannopyranosyl sarsasapogenin, named timosaponin A IV(1) and (5β, 25S)-26-O-β-d-glucopyranosyl-furost-20(22)-en-3,26-diol-3-O-β-d-glucopyranosyl (1 → 4) glucopyranosyl (1 → 2)-β-d-galacopyranoside, named timosaponin B IV(2), were isolated by silica gel column chromatography and preparative HPLC from Anemarrhena asphodeloides Bge. Their structures were elucidated by chemical characters and spectroscopic analysis.

Study on Extraction of Tea Saponin from Camellia oleifera Cake Using Water as Extraction Solvent

This paper studied the effects of liquid-solid ratio, temperature, time and pH value on the extraction rate of tea saponin from the cake of Camellia oleifera seeds by using single factor experiment with the cake of Camellia oleifera seeds as the raw materials, and water as the extraction solvent, and orthogonal test was used to determine the optimal extraction process conditions. The results showed that the extraction ratio of tea saponin could reach up to 95.50% when the liquidsolid ratio was 11：1, extracting temperature of 80 ℃, extraction time of 6 h, and pH value of 9.

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.

预知子中saponins P_K的HPLC分析

目的分析不同种与不同产地预知子中saponins PK的含量,为预知子质量标准建立提供进一步参考。方法采用高效液相色谱分析,Alltech C18色谱柱,乙腈-水(0.1%磷酸)(29.5∶70.5)为流动相,检测波长203 nm。结果saponins PK的保留时间为14 min,线性范围为0.091～0.455μg(r=0.999 9),加样回收率101.93%。所测样品中saponins PK的含量在0.04%～2.04%。结论该法快捷、简便、准确,可用于预知子的质量评价。

Two New Tirucallane-Type Triterpenoid Saponins from Sapindus mukorossi

Two new tirucallane-type triterpenoid saponins, Sapimukoside A (1) and Sapimukoside B (2), were isolated from the roots of Sapindus mukorossi Gaertn. On the basis of spectroscopic and chemical evidence, their structures were elucidated to be 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-arabinopyranosyl-(1-->3)]-beta-D-glucopyranosyl-21, 23R-epoxyl tirucall-7, 24R-diene-3beta, 21-diol (1) and 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyl-21, 23R-epoxyl tirucall-7, 24R-diene-3beta, 21-diol (2).

A New Triterpenoid Saponin from Rhaponticum uniflorum (Compositae)

A new triterpenoid saponin, namely unifloroside, together with five known compounds, was isolated from Rhaponticum uniflorum (L.) DC. The structure of the new triterpenoid saponin was elucidated as 3_O_ α_L_arabinopyranosyl_urs_9(11),12_dien_28_oic acid β_D_glucopyranosyl ester mainly by using 1D, 2D_NMR techniques and chemical methods.

Preliminary Identification of Red Pigment and Positive Correlation between the Contents of Red Pigment and Total Saponins of Panax notoginseng Fruits

[Objective] This study aimed to identify red pigment of Panax notoginseng fruits and explore the correlation between pigment content and total saponins of the fruits. [Method] The red pigment of Panax notoginseng fruits was preliminarily identi- fied with specific color reactions and UV-vis spectra, and the contents of the pigment and total saponins were determined via spectrophotometry. [Result] The red hues of the fruits were contributed by anthocyanins and/or the anthocyanidins. The contents of anthocyanins and total saponins of the fruits both decreased along with thinning of the red hues. The content difference of the anthocyanins in fruits with different red hues reached extremely significant level, but that of total saponins just reached significant level. [Conclusion] The red pigment of P. notoginseng fruits is anthocyanins which are of extremely significant positive correlation with total saponins in contents.

Anticancer effects of saponin and saponin–phospholipid complex of Panax notoginseng grown in Vietnam

Objective:To evaluate the antitumor activity both in vitro and in vivo of saponin–phospholipid complex of Panax notoginseng.Methods:The in vitro cytotoxic effect of saponins extract and saponin–phospholipid complex against human lung cancer NCI-H460 and breast cancer cell lines BT474 was examined using MTS assay.For in vivo evaluation of antitumor potential,saponin and saponin–phospholipid complex were administered orally in rats induced mammary carcinogenesis by 7,12-dimethylbenz(a)anthracene,for 30 days.Results:Our data showed that saponin–phospholipid complex had stronger anticancer effect compared to saponin extract.The IC50 values of saponin–phospholipid complex and saponin extract for NCI-H460 cell lines were 28.47 mg/m L and 47.97 mg/mL,respectively and these values for BT474 cells were 53.18 mg/mL and 86.24 mg/mL,respectively.In vivo experiments,administration of saponin,saponin–phospholipid complex and paclitaxel(positive control) effectively suppressed 7,12-dimethylbenz(a) anthracene-induced breast cancer evidenced by a decrease in tumor volume,the reduction of lipid peroxidation level and increase in the body weight,and elevated the enzymatic antioxidant activities of superoxide dismutase,catalase,glutathione peroxidase in rat breast tissue.Conclusions:Our study suggests that saponin extract from Panax notoginseng and saponin–phospholipid complex have potential to prevent cancer,especially breast cancer.

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.

Responses of Photosynthetic Pigment, Phenol and Total Saponin Contents of Panax notoginseng to Supplemental UV-B under Field Conditions

[Objective] The aim of this study was to investigate the content changes and their correlations of the photosynthetic pigment,phenols,including total phenols,total flavonoids and anthocyanins,and total saponins of the one-year-old P.notoginseng plants under supplemental UV-B stress in fields.[Method] The one-year-old plants were irradiated by UV-B in field for 1 min per day,and the plants under the UV-B lamp were regarded as a circle center,achieving an annular leaf-sampling.The photosynthetic pigment,phenols and total saponins of the leaves were determined spectrophotometrically.[Result] With the increase of sampling radius,the supplemental UV-B intensity decreased significantly,the contents of chlorophyll （Chl） a,Chl b,Chl （a＋b）,carotenoid （Car） and total photosynthetic pigment （Chl＋Car） of the leaves increased extremely significantly,the Chl a/b and total phenol content （TPC） decreased extremely significantly,but the Chl （a＋b）/Car changes were not significant.The contents of total flavonoids,anthocyanins and saponins all increased due to the increasing of UV-B,displaying dose effects.The UV-B intensity was positively correlated with the Chl a/b,and negatively with the Chl a,Chl b,Chl （a＋ b）,Car and （Chl＋Car） contents; and the two of TPC,total flavonoid content （TFC）,total anthocyanin content （TAC） and total saponin content （TSC） were positively correlated,all reaching extremely significant level.The UV-B intensity was positively and significantly correlated with the total flavonoid content （TFC）,negatively and significantly with the Chl （a＋b）/Car,and positively and insignificantly with the TPC,TAC and TSC.[Conclusion] For one-year-old plants of P.notoginseng,UV-B can decrease the contents of the Chl a,Chl b,Chl （a＋b）,Car and （Chl＋Car） and increase the Chl a/b and TPC,and,furthermore,induce the increases of the TFC,TAC and TSC in a dose-dependent manner.However,UV-B can hardly change the Chl （a＋b）/Car.The supplemental UV-B of well-suited dose might be one of the effective measures to improve the TSC of P.notoginseng.

Screening and functional evaluation of the glucose-lowering active compounds of total saponins of Baibiandou(Lablab Semen Album)

Objective To screen forα-glucosidase inhibitor active compounds in the total saponins of Baibiandou(Lablab Semen Album)based on UHPLC-Q-Exactive Orbitrap MS technology and to evaluate its hypoglycemic activity in vivo.Methods Acarbose was used as the positive control,and the median inhibitory concentration(IC50)was used as the evaluation index ofα-glucosidase inhibitory activity to establish an in vitroα-glucosidase inhibition model.Further,UHPLC-Q-Exactive Orbitrap MS technology was used to screen and identify the active compounds ofα-glucosidase inhibitors in the total saponins of Baibiandou(Lablab Semen Album)in order to further verify the activity of the main active monomer and to perform homologous modeling and molecular docking of yeast-derivedα-glucosidase and human-derivedα-glucosidase,while the hypoglycemic activity was evaluated in diabetic mice.Results This study successfully identified 15 compounds with potentialα-glucosidase inhibitory activity,including Chikusetsusaponin IVa,from the total saponins of Baibiandou(Lablab Semen Album).Simultaneously,we verified the activity of the main active monomer Chikusetsusaponin IVa,and showed that it has strongα-glucosidase inhibitory activity.Theα-glucosidase inhibitory concentration IC50 was(565.2±1.026)μg/m L,and the IC50 of acarbose,which was lower than the positive control,was(706.6±1.058)μg/m L.The docking energies of Chikusetsusaponin IVa were–6.1 and–7.7 kcal/mol with yeast-derivedα-glucosidase and human-derivedα-glucosidase molecules,respectively.Both showed strong binding activity,and the levels of alanine aminotransaminase(ALT),aspartate aminotransaminase(AST),UREA,Creatinine(CREA),and cholesterol(CHO)were significantly decreased by Chikusetsusaponin IVa(P<0.05).In addition,it could repair damaged liver and pancreas cells of diabetic mice to some extent.Conclusion This study provides a basis for screeningα-glucosidase inhibitors and structural modifications of the total saponins of Baibiandou(Lablab Semen Album).

A New Triterpenoid Saponin: Se-saponin A

A new triterpenoid saponin, se-saponin A, was isolated from the aerial parts of Clerodendrum serratum var. amplexifolium Moldenke. Its structure was determined by spectral and chemical methods. Only one triterpenoid saponin has been isolated from Clerodendrum genus before. Se-saponin A is the second instance.

New Triterpenoid Saponins from Polycarpon prostratum (Caryophyllaceae)

Three new triterpenoid saponins, namely prostratosides A-C (1-3), were isolated from the whole plant of Polycarpon prostratum (Forssk.) Aschers. et Schwein. ex Aschers. By spectroscopic methods, their structures were determined as 3_O_{β_D_xylopyranosyl_(1→2)_β_D_glucopyranosyl_(1→4)_[β_D_glucopyranosyl_(1→2)]_α_L_arabinopyranoside}_22α_acetoxy_saikogenin G, 3_O_{β_D_xylopyranosyl_(1→2)_β_D_glucopyranosyl_(1→4)_[β_D_glucopyranosyl_(1→2)]_α_L_arabinopyranoside}_16α_hydroxy_22α_acetoxy_saikogenin E and 3_O_{β_D_xylopyranosyl_(1→2)_β_D_glucopyranosyl_(1→4)_[β_D_glucopyranosyl_(1→2)]_α_L_arabinopyranoside}_21β_acetoxy_saikogenin G, respectively.

Ypsilandrosides U-Y,five new steroidal saponins from Ypsilandra thibetica

Phytochemical reinvestigation on the whole plants of Ypsilandra thibetica obtained four new spirostanol glycosides,named ypsilandrosides U-X(1-4),and one new cholestanol glycoside,named ypsilandroside Y(5).Their structures have been established by extensive spectroscopic data and chemical methods.Among them,compound 4 is a rare spirostanol glycoside which possesses a novel 5(6→7)abeo-steroidal aglycone,while compound 1 is a first spiro-stanol bisdesmoside attached to C-3 and C-12,respectively,isolated from the genus Ypsilandra.The induced platelet aggregation activity of the isolates was tested.