Development of 50 InDel-based barcode system for genetic identification of tartary buckwheat resources.

Tartary buckwheat (Fagopyrum tataricum Gartn.) is a highly functional crop that is poised to be the target of many future breeding efforts. The reliable ex situ conservation of various genetic resources is essential for the modern breeding of tartary buckwheat varieties. We developed PCR-based co-dominant insertion/deletion (InDel) markers to discriminate tartary buckwheat genetic resources. First, we obtained the whole genome from 26 accessions across a superscaffold-scale reference genome of 569.37 Mb for tartary buckwheat cv. "Daegwan 3-7." Next, 171,926 homogeneous and 53,755 heterogeneous InDels were detected by comparing 26 accessions with the "Daegwan 3-7" reference sequence. Of these, 100 candidate InDels ranging from 5-20 bp in length were chosen for validation, and 50 of them revealed polymorphisms between the 26 accessions and "Daegwan 3-7." The validated InDels were further tested through the assessment of their likelihood to give rise to a single or a few PCR products in 50 other accessions, covering most tartary buckwheat genome types. The major allele frequencies ranged from 0.5616 at the TB42 locus to 0.9863 at the TB48 locus, with the average PIC value of 0.1532 with a range of 0.0267-0.3712. To create a user-friendly system, the homology of the genotypes between and among the accessions were visualized in both one- (1D) and two-dimensional (2D) barcode types by comparing amplicon polymorphisms with the reference variety, "Daegwan 3-7." A phylogenetic tree and population structure of the 76 accessions according to amplicon polymorphisms for the 50 InDel markers corresponded to those using non-synonymous single nucleotide polymorphism variants, indicating that the barcode system based on the 50 InDels was a useful tool to improve the reliability of identification of tartary buckwheat accessions in the germplasm stocks.

Ethanol Extract of Potentilla supina Linne Suppresses LPS-induced Inflammatory Responses through NF-κB and AP-1 Inactivation in Macrophages and in Endotoxic mice.

In this study, we investigated the antiinflammatory effects of ethanol extracts of Potentilla. supina Linne (EPS) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages and septic mice. EPS suppressed LPS-induced nitric oxide, prostaglandin E2 , TNF-α, interleukin-6 and interleukin-1ß at production and mRNA levels in LPS-induced RAW 264.7 macrophages. Consistent with these observations, EPS attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by downregulation of their promoter activities. Molecularly, EPS reduced the LPS-induced transcriptional activity and DNA-binding activity of nuclear factor-κB (NF-κB), and this was associated with a decrease of translocation and phosphorylation of p65 NF-κB by inhibiting the inhibitory κB-α degradation and IKK-α/ß phosphorylation. Furthermore, EPS inhibited the LPS-induced activation of activator protein-1 by reducing the expression of c-Fos and c-Jun in nuclear. EPS also suppressed the phosphorylation of mitogen-activated protein kinase, such as p38 mitogen-activated protein kinase and c-Jun N-terminal kinase. In an LPS-induced endotoxemia mouse model, pretreatment with EPS reduced the mRNA levels of inducible nitric oxide synthase, cyclooxygenase-2 and proinflammatory cytokines and increased the survival rate of mice. Collectively, these results suggest that the antiinflammatory effects of EPS were associated with the suppression of NF-κB and activator protein-1 activation and support its possible therapeutic role for the treatment of endotoxemia. Copyright © 2017 John Wiley & Sons, Ltd.

α-Solanine Isolated From Solanum Tuberosum L. cv Jayoung Abrogates LPS-Induced Inflammatory Responses Via NF-κB Inactivation in RAW 264.7 Macrophages and Endotoxin-Induced Shock Model in Mice.

α-Solanine, a trisaccharide glycoalkaloid, has been reported to possess anti-cancer effects. In this study, we investigated the anti-inflammatory effects of α-solanine isolated from "Jayoung" a dark purple-fleshed potato by examining its in vitro inhibitory effects on inducible nitric-oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines in LPS-induced RAW 264.7 macrophages and its in vivo effects on LPS-induced septic shock in a mouse model. α-Solanine suppressed the expression of iNOS and COX-2 both at protein and mRNA levels and consequently inhibited nitric oxide (NO) and prostaglandin E2 (PGE2 ) production in LPS-induced RAW 264.7 macrophages. α-Solanine also reduced the production and mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß) induced by LPS. Furthermore, molecular mechanism studies indicated that α-solanine inhibited LPS-induced activation of nuclear factor-κB (NF-κB) by reducing nuclear translocation of p65, degradation of inhibitory κBα (IκBα), and phosphorylation of IκB kinaseα/ß (IKKα/ß). In an in vivo experiment of LPS-induced endotoxemia, treatment with α-solanine suppressed mRNA expressions of iNOS, COX-2, IL-6, TNF-α, and IL-1ß, and the activation of NF-κB in liver. Importantly, α-solanine increased the survival rate of mice in LPS-induced endotoxemia and polymicrobial sepsis models. Taken together, our data suggest that the α-solanine may be a promising therapeutic against inflammatory diseases by inhibiting the NF-κB signaling pathway. J. Cell. Biochem. 117: 2327-2339, 2016. © 2016 Wiley Periodicals, Inc.

Solanum tuberosum L. cv Jayoung Epidermis Extract Inhibits Mite Antigen-Induced Atopic Dermatitis in NC/Nga Mice by Regulating the Th1/Th2 Balance and Expression of Filaggrin.

Solanum tuberosum L. cv Jayoung (JY) is a potato with dark purple flesh and contains substantial amounts of polyphenols. In this study, we evaluated the therapeutic effects of S. tuberosum L. cv JY in a mouse model of Dermatophagoides farinae body (Dfb)-induced atopic dermatitis (AD). The ethanol extract of the peel of JY (EPJ) ameliorated Dfb-induced dermatitis severity, serum levels of immunoglobulin E (IgE) and thymus and activation-regulated chemokine. Histological analysis of the skin also revealed that EPJ treatment significantly decreased mast cell infiltration. The suppression of dermatitis by EPJ treatment was accompanied by a decrease in the skin levels of type 2 helper T-cell cytokines such as interleukin (IL)-4, IL-5, and IL-13. The induction of thymic stromal lymphopoietin, which leads to a systemic Th2 response, was also decreased in the skin by EPJ. Nuclear translocation of nuclear factor-κB p65 was decreased by EPJ in Dfb-induced NC/Nga mice. The protein expression of filaggrin in the AD-like skin lesions was restored by EPJ treatment. These results suggested that EPJ may be a potential therapeutic tool for the treatment of AD.

α-Chaconine isolated from a Solanum tuberosum L. cv Jayoung suppresses lipopolysaccharide-induced pro-inflammatory mediators via AP-1 inactivation in RAW 264.7 macrophages and protects mice from endotoxin shock.

In this study, we investigated the molecular mechanisms underlying the anti-inflammatory effects of α-chaconine in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages and in LPS-induced septic mice. α-Chaconine inhibited the expressions of cyclooxygenase-2 (COX-2), interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α) at the transcriptional level, and attenuated the transcriptional activity of activator protein-1 (AP-1) by reducing the translocation and phosphorylation of c-Jun. α-Chaconine also suppressed the phosphorylation of TGF-ß-activated kinase-1 (TAK1), which lies upstream of mitogen-activated protein kinase kinase 7 (MKK7)/Jun N-terminal kinase (JNK) signaling. JNK knockdown using siRNA prevented the α-chaconine-mediated inhibition of pro-inflammatory mediators. In a sepsis model, pretreatment with α-chaconine reduced the LPS-induced lethality and the mRNA and production levels of pro-inflammatory mediators by inhibiting c-Jun activation. These results suggest that the anti-inflammatory effects of α-chaconine are associated with the suppression of AP-1, and support its possible therapeutic role for the treatment of sepsis.

Chloroform fraction of Solanum tuberosum L. cv Jayoung epidermis suppresses LPS-induced inflammatory responses in macrophages and DSS-induced colitis in mice.

In this study, the authors investigated the molecular mechanism underlying the antiinflammatory effects of the chloroform fraction of the peel of 'Jayoung' (CFPJ), a color-fleshed potato, on lipopolysaccharide (LPS)-induced RAW 264.7 macrophages and in mice with dextran sulfate sodium (DSS)-induced colitis. CFPJ inhibited the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the transcription level, and attenuated the transcriptional activity of nuclear factor-κB (NF-κB) by reducing the translocation of NF-κB depending on degradation of inhibitory κB-α (IκB-α). Furthermore, CFPJ attenuated the phosphorylations of mitogen-activated protein kinase kinases3/6 (MKK3/6) and of p38. In colitis model, CFPJ significantly reduced the severity of colitis and the productions and protein levels of pro-inflammatory mediators in colonic tissue. These results suggest that the anti-inflammatory effects of CFPJ are associated with the suppression of NF-κB and p38 activation in macrophages, and support its possible therapeutic role for the treatment of colitis.

Pectolinarin and Pectolinarigenin of Cirsium setidens Prevent the Hepatic Injury in Rats Caused by D-Galactosamine via an Antioxidant Mechanism.

To identify the hepatoprotective component from the leaves of Cirsium setidens (Compositae), the methanolic extract was divided into two fractions, chloroform and butanol fractions, and their hepatoprotective efficacy was evaluated in a rat model of hepatic injury caused by D-galactosamine (GalN). Hepatoprotective activity was measured by the activity of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH). Glutathione metabolism was measured via biochemical parameters such as glutathione (GSH), glutathione reductase (GR), gamma-glutamylcysteine synthetase (GCS), glutathione S-transferase (GST), and superoxide dismutase (SOD) levels. We subjected the butanol fraction, which had higher activity, to column chromatography to yield pectolinarin, which was further hydrolyzed to yield pectolinarigenin. Administration (10, 20 mg/kg, p.o.) of the main flavonoid glycoside component, pectolinarin, and its aglycone, pectolinarigenin, for 2 weeks significantly decreased the activity levels of AST, ALT, ALP and LDH, indicating that the two compounds have hepatoprotective activity. Pectolinarin and pectolinarigenin also increased activity levels of GSH, GR, GCS, and GST, as well as SOD. The significant effect was only seen in SOD activity. This suggests that the two components exhibit hepatoprotective activity mainly via SOD antioxidant mechanism.

Anti-ulcerogenic effects of the flavonoid-rich fraction from the extract of Orostachys japonicus in mice.

Orostachys japonicus (Crassulaceae) herbal preparations have been used to treat gastric ulcer or gastric cancer disease in Korean folk medicine. To demonstrate the effects of the methanol (MeOH) extract of O. japonicus and its fractions on gastric lesions and pain, the MeOH extract was fractionated into triterpene-rich and flavonoid-rich (FRF) fractions. Second, the fractions were subjected to analgesic assays including hot plate and writhing assays and anti-ulcerogenic assays in HCl/ethanol-induced- and indomethacin/bethanechol-induced ulcer models in mice. In this experiment, it was found that the FRF most significantly reduced ulcerative indices and pain in mice, although the MeOH extract was also effective. Oral administration of the FRF highly reduced the diameter of gastric lesion induced by HCl/ethanol (inhibitory effect, 53%) and by indomethacin/bethanechol (inhibitory effect, 36%) at the 100 mg/kg dose. In addition, oral administration of 200 mg/kg FRF markedly increased the reaction time in the hot plate test by 52% and decreased stretching episodes (45%) in the writhing test. These results suggest that the active component of O. japonicus exhibiting the potent anti-ulcerative and antinociceptive effect is included in the FRF. The anti-ulcerative effects of the MeOH extract and the FRF were also supported by gastric juice and gastric acid volumes and pH in pylorus-ligated mice. Taken together, these results provide evidence-based support for the traditional use of O. japonicus for gastric disease.

The MeOH extract of Pleurospermum kamtschaticum and its active component buddlejasaponin (IV) inhibits intrinsic and extrinsic hyperlipidemia and hypercholesterolemia in the rat.

The inhibitory effect of the MeOH extract of Pleurospermum kamtschaticum (Umbelliferase) and its fractions were tested in hyperlipidemic and hypercholesterolemic rats using four animal models induced using poloxamer-407 or using Triton WR-1339 as intrinsic inducers and by 30% corn oil or high cholesterol diet as extrinsic inducers. We measured serum triglyceride, total cholesterol, HDL-cholesterol and LDL-cholesterol levels as measures of its hypocholesterolemic or hypolipidemic effects. Since the MeOH extract and the BuOH fraction of Pleurospermum kamtschaticum were found to be active using these four hypolipidemic assays, its major saponin buddlejasaponin IV {BS(IV)} isolated from the BuOH fraction were also tested to demonstrate the active components. BS(IV) was found to significantly inhibit hypercholesterolemia and hyperlipidemia by extrinsic and intrinsic inducers. In particular, BS(IV) reduced the blood thiobarbituric acid reactive substance (TBARS) and hydroxy radical levels, and increased superoxide dismutase activity in high cholesterol diet-induced rats, thus suggesting that BS(IV) reduces oxidative stress caused by a high cholesterol diet. Moreover, these effects of BS(IV) were comparable to probucol, which was used as a positive control. These results suggested that Pleurospermum kamtschaticum which is traditionally used to treat atherosclerosis and its active major saponin BS(IV) could be used to treat hypercholesterolemia or hyperlipidemia.

Anti-inflammatory activities of ent-16alphaH,17-hydroxy-kauran-19-oic acid isolated from the roots of Siegesbeckia pubescens are due to the inhibition of iNOS and COX-2 expression in RAW 264.7 macrophages via NF-kappaB inactivation.

To isolate the anti-inflammatory components in Siegesbeckia pubescens root, we performed activity-guided fractionation using a carrageenan-induced edema rat model. Antinociceptive effects were followed using acetic acid-induced abdominal constriction and hot plate tests in mice. Chloroform extract was subjected to silica gel and octadesyl silane (ODS) column chromatography, and a diterpene was isolated which was identified as ent-16alphaH,17-hydroxy-kauran-19-oic acid (siegeskaurolic acid). Pretreatment with siegeskaurolic acid (20 or 30 mg/kg/day, p.o.) exhibited anti-inflammatory and antinociceptive effects in these animal models. To investigate the mechanisms underlying this anti-inflammatory action, we investigated the effect of siegeskaurolic acid on lipopolysaccharide (LPS)-induced responses in a murine macrophage cell line, RAW 264.7. Siegeskaurolic acid was found to significantly inhibit the productions of nitric oxide (NO), prostaglandin E(2) (PGE(2)), and tumor necrosis factor-alpha (TNF-alpha). Consistent with these findings, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins, and iNOS, COX-2, and TNF-alpha mRNAs were found to be inhibited by siegeskaurolic acid. Furthermore, siegeskaurolic acid inhibited the nuclear factor-kappaB (NF-kappaB) activation induced by LPS, and this was associated with the prevention of inhibitor kappaB degradation (I kappaB), and subsequently with decreased nuclear p65 and p50 protein levels. Taken together, our data indicate that the anti-inflammatory and antinociceptive properties of siegeskaurolic acid may be due to iNOS, COX-2 and TNF-alpha inhibition via the down-regulation of NF-kappaB binding activity.

The anti-gastropathic and anti-rheumatic effect of niga-ichigoside F1 and 23-hydroxytormentic acid isolated from the unripe fruits of Rubus coreanus in a rat model.

This study was undertaken to produce the clinical merits of two natural antinociceptive anti-inflammatory triterpenoids which synthetic anti-inflammatory drugs do not have. The triterpenoid glycoside niga-ichigoside F1 (NIF1) and its aglycone 23-hydroxytormentic acid (23-HTA), which were isolated from the unripe fruits of Rubus coreanus (Rosaceae), reduced rheumatoid arthritis (RA) factor and C-reactive protein (CRP) factor in Freund's complete adjuvant reagent-induced rats, suggesting that these two triterpenoids had an anti-rheumatic effect. It was also shown that treatment with NIF1 or 23-HTA reduced gastric lesion extent, acidity and total gastric acid output induced by EtOH plus sodium salicylate in a gastric secretion test. Moreover, 23-HTA had a greater effect than the glycoside, NIF1. To clarify the anti-gastropathic mechanism of these two compounds, their free radical scavenging activities in the gastric mucosa were examined in a rat EtOH-sodium salicylate-induced gastropathy model. The two compounds significantly increased superoxide dismutase and glutathione peroxidase activities, indicating that the healing effects of NIF1 and 23-HTA against gastropathy are associated with free radical scavenging enzyme activities. These results support the notion that the long-term administration of NIF1 or 23-HTA should overcome the adverse effects of synthetic anti-inflammatory drugs.

Antinociceptive anti-inflammatory effect of Monotropein isolated from the root of Morinda officinalis.

The root of Morinda officinalis (Rubiaceae) is used to treat rheumatoid arthritis and impotence in the traditional Oriental medicine. To identify the antinociceptive anti-inflammatory components of this crude drug, we adopted an activity-directed fractionation approach. The active fraction of the BuOH extract of M. officinalis root was subjected to silica gel and ODS column chromatography to yield two diterpenes, compounds 1 and 2 and these were identified as monotropein and deacetylasperulosidic acid, respectively. The iridoid glycoside, monotropein, was tested for its anti-inflammatory antinociceptive effects using hot plate- and writhing antinociceptive assays and by using carrageenan-induced anti-inflammatory assays in mice and rats. Pretreatment with monotropein (at 20, 30 mg/kg/d, p.o.) significantly reduced stretching episodes and prolonged action time in mice. It also significantly reduced acute paw edema by carrageenan in rats. These results indicate that monotropein contributes to the antinociceptive and anti-inflammatory action of Morinda officinalis root.

Isolation of saponins with the inhibitory effect on nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha production from Pleurospermum kamtschaticum.

As an attempt to search for bioactive natural products exerting antiinflammatory activity, we have isolated two saponins were isolated from the aerial portion of Pleurospermum kamtschaticum (Umbelliferae) by nitrite assay activity-directed chromatographic fractionation. They were identified as saikogenin F 3-O-{beta-D-glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->3)]-beta-D-fucopyranoside} (buddlejasaponin IV, 1) and 3beta,16beta,23,28-tetrahydroxy-11alpha-methoxyolean-12-ene 3-O-{beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->3)]-beta-D-fucopyranoside} (buddlejasaponin IVa, 2). Compound 1 significantly inhibited nitric oxide (NO) production, and it also significantly decreased prostaglandin E2 (PGE2) and tumor necrosis factor-alpha (TNF-alpha) release in the lipopolysaccharide (LPS)-activated macrophage Raw 264.7 cells whereas compound 2 was much less active. Saikogenin A (3) and -H (4) were obtained by hydrolyzing 1 and 2. Although these sapogenin showed strong NO inhibition, these effects were caused by the cytotoxic effect on Raw 264.7 cells. These results supported the notion that buddlejasaponin IV is a major inhibitors of NO, PGE2 and TNF-alpha production in P. kamtschaticum.

Eugenol isolated from the essential oil of Eugenia caryophyllata induces a reactive oxygen species-mediated apoptosis in HL-60 human promyelocytic leukemia cells.

Eugenol is a major component of essential oil isolated from the Eugenia caryophyllata (Myrtaceae), which has been widely used as a herbal drug. In this study, we investigated the effects of eugenol on the cytotoxicity, induction of apoptosis, and the putative pathways of its actions in human promyelocytic leukemia cells (HL-60) under the standard laboratory illumination. Eugenol-treated HL-60 cells displayed features of apoptosis including DNA fragmentation and formation of DNA ladders in agarose gel electrophoresis. We observed that eugenol transduced the apoptotic signal via ROS generation, thereby inducing mitochondrial permeability transition (MPT), reducing anti-apoptotic protein bcl-2 level, inducing cytochrome c release to the cytosol, and subsequent apoptotic cell death. Taken together, the present study demonstrated that ROS plays a critical role in eugenol-induced apoptosis in HL-60, and this is the first report on the mechanism of the anticancer effect of eugenol.

In-vitro and in-vivo anti-inflammatory and antinociceptive effects of the methanol extract of the roots of Morinda officinalis.

The anti-inflammatory effects of the methanol extract of the roots of Morinda officinalis (MEMO) (Rubiaceae) were evaluated in-vitro and in-vivo. The effects of MEMO on lipopolysaccharide (LPS)induced responses in the murine macrophage cell line RAW 264.7 were examined. MEMO potently inhibited the production of nitric oxide (NO), prostaglandin E2 and tumour necrosis factor-alpha (TNF-alpha) in LPS-stimulated RAW 264.7 macrophages. Consistent with these results, the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein level, and of iNOS, COX-2 and TNF-alpha at the mRNA level, was also inhibited by MEMO in a concentration-dependent manner. Furthermore, MEMO inhibited the nuclear factor kappa B (NF-kappaB) activation induced by LPS, and this was associated with the prevention of degradation of the inhibitor kappaB (IkappaB), and subsequently with attenuated p65 protein in the nucleus. The anti-inflammatory effect of MEMO was examined in rats using the carrageenan-induced oedema model. The antinociceptive effects of MEMO were assessed in mice using the acetic acid-induced abdominal constriction test and the hot-plate test. MEMO (100, 200 mg kg-1 per day, p.o.) exhibited anti-inflammatory and antinociceptive effects in these animal models. Taken together, the data demonstrate that MEMO has anti-inflammatory and antinociceptive activity, inhibiting iNOS, COX-2 and TNF-alpha expression by down-regulating NF-kappaB binding activity.

Antiinflammatory effects of chiisanoside and chiisanogenin obtained from the leaves of Acanthopanax chiisanensis in the carrageenan- and Freund's complete adjuvant-induced rats.

To find the antiinflammtory constituents of Acanthopanax chiisanensis (Araliaceae) leaves, phytochemical isolation procedures were performed by activity-guided fractionation in carrageenan- and Freund's complete adjuvant (FCA) reagent-induced rat models, respectively. In the two assay system, the MeOH extract (100 and 250 mg/kg, p.o.) showed significant antiinflammtory effects. Since BuOH extract among the fractionated extracts exhibited the most potent effect, it was subjected to column chromatography to yield a main triterpene glycoside, chiisanoside (1). This compound was hydrolyzed in alkaline solution to find the biological activity of produced aglycone, chiisanogenin (1a). Oral treatment with compounds 1 and 1a produced significant antiinflammtory effects at 10 and 30 mg/kg dose, and 1a was more potent than 1. The antiiflammtory effects of the two compounds were supported by the reduction of carrageenan-induced lipid peroxidation and hydroxy radical in serum. Furthermore, treatment with 1 and 1a significantly reduced rheumatoid arthritis (RA) and C-reactive protein (CRP) factors in the rat induced by Freund's complete adjuvant reagent. Compounds, 1 and 1a, inhibited xanthine oxidase activity and increased superoxide dismutase (SOD), glutathione peroxidase and catalase indicating that both compounds scavenged reactive oxygen species (ROS).

19Alpha-hydroxyursane-type triterpenoids: antinociceptive anti-inflammatory principles of the roots of Rosa rugosa.

To search for antiinflammtory 19alpha-hydroxyursane-type triterpenoids, the MeOH extract of the roots of Rosa rugosa (Rosaceae) was fractionated. The active fraction of the EtOAc extract was hydrolyzed in alkaline solution to give a hydrolyzed fraction. Both extracts showed antiinflammatory/antinociceptive action in acetic acid-induced writhing and hot plate testing and in a carrageenan-induced paw edema model in mice and rats. Repeated chromatography of the EtOAc extract on both silica gel and octadecylsilane columns led to the isolation of kaji-ichigoside F1 (1, euscaphic acid 28-O-glucoside) and rosamultin (2, tormentic acid 28-O-glucoside). The hydrolyzed fraction was also subjected to silica gel column and octadecylsilane column chromatography to produce euscaphic acid (3) and tormentic acid (4). The potencies were observed in the following order: 4>3>2>1. These results suggest that 19alpha-hydroxyursane-type triterpenoids are responsible for the antiinflammatory/antinociceptive action of R. rugosa roots.

Structure of a new echinocystic acid bisdesmoside isolated from Codonopsis lanceolata roots and the cytotoxic activity of prosapogenins.

We isolated a new saponin named codonoposide (1) from the roots of Codonopsis lanceolata (Campanulaceae) and characterized it as 3-O-[beta-D-xylopyranosyl(1-3)-beta-D-glucuronopyranosyl]-3beta,16alpha-dihydroxyolean-28-oic acid 28-O-[beta-D-xylopyranosyl (1-3)-alpha-L-rhamnopyranosyl (1-2)-alpha-L-arabinopyranosyl] ester by chemical, physicochemical, and 2DNMR techniques. Complete hydrolysis of 1 produced a sapogenin (1a), and the partial hydrolysis and further isolation afforded two prosapogenins (1b, 1c). The structures of 1a, 1b, and 1c were found to be 3beta,16alpha-dihydroxyolean-28-oic acid (echinocystic acid, 1a), 3-O-beta-D-glucuronopyranoside of 1a, and 3-O-beta-D-xylopyranosyl (1-3)-beta-D-glucuronopyranoside of 1a, respectively, on the basis of spectroscopic data. On MTT assay, 1a showed marginal cytotoxic activity whereas 1b exhibited more cytotoxicity than 1a. However, the bisdesmosylsaponin 1 exhibited no cytotoxicity (IC(50)>0.3 mM against tested cell lines). This result indicated that glycoside linkage of glucuronic acid at C-3 enhances the cytotoxicity of sapogenin (1a), and additive glycosylation of xylose to 1b strongly enhances the cytotoxicity of 3-O-monosaccharides (1b). Therefore, true forms of codonoposide for the cytotoxicity must be sapogenins or prosapogenins.