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.

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.

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。

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.

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).

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.

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.

Two Novel Antifungal Saponins from Tibetan Herbal Medicine Clematis tangutica

Antifungal assay-guided isolation of the ethanol extract of the aerial parts of Clematis tangutica yielded two novel triterpene saponins. Their structures were determined to be 3-O-a-L-arabinopyranosyl hederagenin 28-O-a-L-rhamnopyranosyl ester (1) and 3-O-b-D- glucopyranosyl(14)-a-L-arabinopyranosyl hederagenin 28-O-a-L-rhamnopyranosyl ester (2) on the basis of spectral data and chemical reactions.

Adjuvant effects of saponins on animal immune responses

Vaccines require optimal adjuvants including immunopotentiator and delivery systems to offer long term protection from infectious diseases in animals and man. Initially it was believed that adjuvants are responsible for promoting strong and sustainable antibody responses. Now it has been shown that adjuvants influence the isotype and avidity of antibody and also affect the properties of cell-mediated immunity. Mostly oil emulsions, lipopolysaccharides, polymers, saponins, liposomes, cytokines, ISCOMs (immunostimulating complexes), Freund’s complete adjuvant, Freund’s incomplete adjuvant, alums, bacterial toxins etc., are common adjuvants under investigation. Saponin based adjuvants have the ability to stimulate the cell mediated immune system as well as to enhance antibody production and have the advantage that only a low dose is needed for adjuvant activity. In the present study the importance of adjuvants, their role and the effect of saponin in immune system is reviewed.

Two Triterpenoid Saponins from Lonicera Japonica

A new triterpenoid saponin, together with a known saponin. was isolated from the flowers of Lonicera Japonica Thunb. Using chemical and spectroscopic methods, mainly 2D NMR technique, their structures were deduced to be 3-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 3)-alpha-L-rhamnospyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl-(l --> 6)-beta-D-glucopyranoside and 3-alpha-L-rhamnospyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-alpha-L-rhamnospyranosyl- (1 --> 2)-[beta-D-xylcopyranosyl-( 1 --> 6)]-beta-D-glucopyranoside, respectively.

Inhibitory Effects of Saponins From Anemarrhena asphodeloides Bunge on the Growth of Vascular Smooth Muscle Cells

Objective To investigate the effects of saponins from Anemarrhena asphodeloides Bunge （SAaB） （Botanical Name： Anemarrhena Asphodeloidis Rhizoma） on the growth of vascular smooth muscle cells （VSMCs）. Methods Cell proliferation was measured by a newly developed cell proliferation reagent, WST-1. Cell apoptosis was assayed by flow cytometry through detecting annexin V. Nitric oxide production was evaluated using confocal laser scanning microscopy with diaminofluorescein diacetate （DAF-2, DA）. Cell aldose reductase （AR） activity, as well as the effect of Epalrestat and interleukin-1β were also explored. Results WST assay showed that cell proliferation induced by serum was significantly inhibited by SAaB （P〈0.01）. Flow cytometry analysis revealed that SAaB could enhance apoptotic rate of VSMCs （P〈0.01）. Nitric oxide production was significantly enhanced after administration of SAaB and interleukin-Iβ Moreover, AR activity of VSMCs was also remarkably inhibited by both SAaB and Epalrestat （P〈 0.01）. Conclusion SAaB can inhibit proliferation and enhance apoptosis of VSMCs. It may protect vascular cells by inhibiting VSMC proliferation and augmenting apoptotic rate of VSMCs via NO-dependent pathway.

Panax notoginseng saponins preconditioning protects rat liver grafts from ischemia/reper- fusion injury via an antiapoptotic pathway

BACKGROUND: Ischemia/reperfusion (I/R) injury is a major cause of primary graft dysfunction and renders an al- lograft more immunogenic in orthotopic liver transplanta- tion (OLT). Panax notoginseng saponins (PNS) has been re- ported to exert protective effects against I/R injury to vari- ous organs. The objective of this study is to investigate whether PNS preconditioning protects rat liver grafts from I/R injury via an antiapoptotic pathway. METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation ( OLT) and were divided into PNS preconditioning group (group P) and normal saline control group (group N) randomly according to whether PNS (50 mg/kg) was injected intra- venously 1 hour before liver grafts harvesting, and sham group (group S). The animals were separately killed 2, 6 and 24 hours after reperfusion. Plasma samples were collect- ed for test of alanine amino-transferase (ALT) and aspartate aminotransferase (AST). Liver tissues were collected to de- tect histological changes, apoptosis and the expression of TNF-α, Bcl-2 and Caspase-3 mRNA. RESULTS: The serum levels of ALT and AST and the apop- tosis index (AI) of liver tissue in group P were lower than in group N significantly 2, 6 and 24 hours after reperfusion. Compared with group N, the expression of TNF-a and Caspase-3 mRNA was reduced significantly in group P 2 and 6 hours after reperfusion and the expression of Bcl-2 mRNA was enhanced significantly in group P 6 and 24 hours after reperfusion. CONCLUSIONS: PNS preconditioning protects liver grafts from I/R injury effectively in rat OLT via an antiapoptotic pathway. The antiapoptotic mechanisms of PNS may in- clude inhibiting the expression of TNF-a and Caspase-3 and enhancing the expression of Bcl-2.

Two New Oleanene-Type Triterpenoid Saponins from Pueraria peduncularis

From the radix of Pueraria peduncularis Grah., two new oleanene-type triterpenoid saponins named pedunsaponins B (2) and C (3) were isolated. Their structures were determined as 3-O-(6-O-methyl)-beta-D-glucuronopyranosyl-3 beta.15 alpha-diol-12-oleanene-16-one (2) and 3-O-beta-D-glucopyranosyl-( 1 --> 3)-beta-D-glucuronopyranosyl-3 beta, 15 alpha-diol-12-oleanene-16-one (3) on the basis of spectroscopic evidence and chemical reactions.

Effects of total saponins of Panax notoginseng on immature neuroblasts in the adult olfactory bulb following global cerebral ischemia/reperfusion

The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.

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.

Two new saponins from the aerial part of Astragalus membranaceus var. mongholicus

Two new saponins named mongholicoside A （1） and mongholicoside B （2） were isolated from the aerial part of Astragalus membranaceus var mongholicus. Their structures were determined by 1D and 2D NMR, ESI-MS techniques and chemical methods.

Panax notoginseng saponins influence on transplantation of neural stem cell-derived dopaminergic neurons in a rat model of Parkinson’s disease

BACKGROUND： Dopaminergic neurons differentiated from neural stem cells have been successfully used in the treatment of rat models of Parkinson＇s disease; however, the survival rate of transplanted cells has been low. Most cells die by apoptosis as a result of overloaded intracellular calcium and the formation of oxygen free radicals. OBJECTIVE： To observe whether survival of transplanted cells, transplantation efficacy, and dopaminergic differentiation from neural stem cells is altered by Panax notoginseng saponins （PNS） in a rat model of Parkinson＇s disease. DESIGN, TIME AND SETTING： Cellular and molecular biology experiments with randomized group design. The experiment was performed at the Animal Experimental Center, First Hospital of Sun Yat-sen University from April to October 2007. MATERIALS： Thirty-two adult, healthy, male Sprague Dawley rats, and four healthy Sprague Dawley rat embryos at gestational days 14-15 were selected. The right ventral mesencephalon was injected with 6-hydroxydopamine to establish a model of Parkinson＇s disease. 6-hydroxydopamine and apomorphine were purchased from Sigma, USA. METHODS： Neural stem cells derived from the mesencephalon of embryonic rats were cultivated and passaged in serum-free culture medium. Lesioned animals were randomly divided into four groups （n = 8）： dopaminergic neuron, dopaminergic neuron ＋ PNS, PNS, and control. The dopaminergic neuron group was injected with 3 μL cell suspension containing dopaminergic neurons differentiated from neural stem cells. The dopaminergic neurons ＋ PNS group received 3 μ L dopaminergic cell suspension combined with PNS （250 mg/L）. The PNS group received 3 μL PNS （250 mg/L）, and the control group received 3 μL DMEM/F12 culture medium. MAIN OUTCOME MEASURES： The rats were transcardially perfused with 4% paraformaldehyde at 60 days post-grafting for immunohistochemistry. The rats were intraperitoneally injected with apomorphine （0.5 mg/kg） to induce rotational behavior. RESULTS： Cell counts of tyrosine hydroxylase-positive neurons in the dopaminergic neuron ＋ PNS group were （732±82.6） cells/400-fold field. This was significantly greater than the dopaminergic neuron group [（326 ± 34.8） cells/400-fold field, P 〈 0.01]. Compared to the control group, the rotational asymmetry of rats that received dopaminergic neuron transplants was significantly decreased, beginning at 20 days after operation （P 〈 0.01）. Rotational asymmetry was further reduced between 10-60 days post-surgery in the dopaminergic neuron ＋ PNS group, compared to the dopaminergic neuron group （P 〈 0.01）. CONCLUSION： Panax notoginseng saponins can increase survival and effectiveness of dopaminergic neurons differentiated from neural stem cells for transplantation in a rat model of Parkinson＇s disease.

Effects of total soy saponins on free radicals in the quadriceps femoris,serum testosterone,LDH,and BUN of exhausted rats

Purpose:The aim of this study was to investigate the impact of total soy saponins(TS) on the free radical metabolism from the quadriceps femoris muscle,serum testosterone,lactate dehydrogenase(LDH),and blood urea nitrogen(BUN) in rats exercised to exhaustion.Methods:A one-time exhausted treadmill exercise session was used.Sprague-Dawley rats were divided into 4 groups:a control group—animals receiving no TS and no exercise(NTSNE),animals receiving TS but no exercise group(TSNE),animals receiving no TS but exercised to exhaustion group(NTSE),and animals receiving TS and exercised to exhaustion group(TSE).The TSNE and TSE groups were fed TS at a dosage of 20 mg/kg body weight once per day for 2 weeks.The NTSE group was given a placebo,and the NTSNE group was not given any treatment.The NTSE and TSE groups were exercised at speed of 30 m/min on treadmill until exhausted.The exercise time and exercise distance were recorded when the rats became exhausted and the rats were then decapitated and anatomized immediately.A 10% homogenate of the quadriceps femoris tissue was prepared.The levels of superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),glutathione peroxidase(GSH-Px),glutathione reductase(GR),reduced glutathione(GSH),total antioxidant capacity(T-AOC),LDH,BUN,and serum testosterone were tested.Results:TS significantly increased the exercise time to exhaustion by 20.62%(p < 0.05).The MDA levels were decreased significantly in the TSNE group than in NTSNE group(p < 0.05);the T-AOC levels increased significantly in the TSNE group than in the other 3 groups(p < 0.01,p < 0.05,p < 0.05).The LDH activity significantly increased in the NTSE group than in TSNE group(p < 0.05).The BUN levels significantly increased in the NTSE group than in the other 3 groups(p < 0.01,p < 0.01,p < 0.05),and significantly increased in the TSE group than in NTSNE and TSNE groups(both p < 0.01).The serum testosterone levels increased significantly in the TSNE group than in the other 3 groups(all p < 0.01).SOD,CAT,GSH-Px,GR,and GSH were not statistically different among the groups.Conclusion:TS can significantly improve the exercised rats' serum testosterone level and antioxidant activity in their quadriceps femoris to varying degrees,decrease MDA and serum LDH and BUN levels,increase the exercise time,and delay the occurrence of the fatigue.

Neuroprotective effects of total saponins from Rubus parvifolius L. on cerebral ischemia/reperfusion injury in rats

This study examines the neuroprotective effects and mechanisms of action of total saponins from Rubus parvifolius L. （TSRP） on focal cerebral ischemia and reperfusion injury in rats. Focal cerebral ischemia and reperfusion injury was performed in rats using the suture method. The results indicate that intragastric injection of TSRP, at 5, 10 and 20 mg/kg, could decrease neurological impairment, reduce cerebral infarct volume, diminish pathological changes, and significantly inhibit the apoptosis of neurons surrounding the ischemic area. In addition, TSRP upregulated the expression of the anti-apoptotic factor Bcl-2, at the protein and mRNA levels, and it downregulated the expression of the pro-apoptotic factor Bax, at the protein and mRNA levels. These findings indicate that TSRP protects against cerebral ischemia/reperfusion injury, and that it may do so by regulating the expression of Bcl-2 and Bax.