• Korean Journal of Pharmacognosy
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• v.48 no.4
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• pp.261-267
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• 2017

As part of the search for bioactive constituents of Korean medicinal plants, twelve steroids (1-12) were isolated from the rhizomes of Dioscorea nipponica. The isolated compounds were identified as diosgenin ($3{\beta}$, 25R)-spirost-5-en-3-ol (1), 25(R)-dracaenoside E (2), dioscin (3), gracillin (4), prosapogenin B (5), 25(R)-dracaenoside G (6), diosgenin 3-O-${\beta}$-D-glucopyranosyl($1{\rightarrow}3$)-${\beta}$-D-glucopyranoside (7), ophipogonin C′ (8), 7-oxodioscin (9), protodioscin (10), hypoglaucin F (11), and protoneogracillin (12). Their structures were characterized by spectroscopic data and identified by comparing these data with those in the literatures. All the isolates (1-12) were evaluated for their neuroprotective effects through induction of nerve growth factor in C6 glioma cells and effects on nitric oxide (NO) production in murine microglia cell line BV-2. Compounds 7 and 12 were found to induce upregulation of NGF secretion without causing significant cell toxicity and compound 4 exhibited potent anti-neuroinflammatory activity.

• Archives of Pharmacal Research
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• v.24 no.1
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• pp.44-50
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• 2001

To search for cytotoxic components from Allium victoriallis , MTT assays on each extract and an isolated component, gitogenin 3-O-lycotetroside, were performed against cancer cell lines. Cytotoxicities of most extract were shown to be comparatively weak, though $IC_50$ values of $CHCl_3$fraction was found to be <31.3-368.4 $\mu\textrm{g}/ml$. From the incubated methanol extract at $36^{\circ}C, eleven kinds of organosulfuric flavours were predictable by CG-MS performance. The most abundant peak was revealed to be 2-vinyl-4H-1,3-dithiin(1) by its mass spectrum. Further, this extract showed significant cytotoxicities toward cancer cell lies. Silica gel column chromatography of the n-butanol fraction led to the isolation of gitogenin 3-O-lycotetroside (3) along with astragalin (4) and kaempferol 3, 4'-di-O-$\beta$-D-glycoside (5). This steroidal saponin exhibited significant cytotoxic activities ($IC_50$, 6.51-36.5 $\mu\textrm{g}/ml$) over several cancer cell lines. When compound 3 was incubated for 24 h with human intestinal bacteria, a major metabolite was produced and then isolated by silica gel column chromatography. By examining parent and prominent ion peak in FAB-MS spectrum of the metabolite, the structure was speculated not to be any of prosapogenins of 3, suggesting that spiroketal ring were labile to the bacterial reaction. These suggest that disulfides produced secondarily are the antitumor principles.

• Journal of Ginseng Research
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• v.44 no.1
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• pp.145-153
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• 2020

Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions, especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarily on its roots, whereas the seeds remain poorly understood. This study explores the phytochemical and biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using various chromatographic methods. The chemical structures of the isolates were determined by spectroscopic data. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin, pseudoginsenoside RT₈, along with 15 known compounds. Pseudoginsenoside RT₈ exhibited more potent antiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction of proinflammatory genes such as interleukin-1β, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species and nitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT₈ has a pharmaceutical potential as an antiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novel bioactive molecules.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6945-6948
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• 2013

Background: Diosgenin, a steroidal saponin from a therapeutic herb, fenugreek (Trigonellafoenum-graceum L.), has been recognized to have anticancer properties. Telomerase activity is not detected in typical healthy cells, while in cancer cell telomerase expression is reactivated, therefore providing a promising cancer therapeutic target. Materials and Methods: We studied the inhibitory effect of diosgenin on human telomerase reverse transcriptase gene (hTERT) expression which is critical for telomerase activity. MTT- assays and qRT-PCR analysis were conducted to assess cytotoxicity and hTERT gene expression inhibition effects, respectively. Results: MTT results showed that $IC_{50}$ values for 24, 48 and 72h after treatment were 47, 44 and $43{\mu}M$, respectively. Culturing cells with diosgenin treatment caused down-regulation of hTERT expression. Discussion: These results show that diosgenin inhibits telomerase activity by down-regulation of hTERT gene expression in the A549 lung cancer cell line.

• Mass Spectrometry Letters
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• v.4 no.3
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• pp.55-58
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• 2013

Dioscin is a biologically active steroidal saponin with anticancer and hepatoprotective effects. A rapid, selective, and sensitive liquid chromatographic method with electrospray ionization tandem mass spectrometry was developed for the quantification of dioscin in rat plasma. Dioscin was extracted from rat plasma using ethyl acetate at acidic pH. The analytes were separated on a Halo C18 column using gradient elution of acetonitrile and 0.1% formic acid and detected by tandem mass spectrometry in selected reaction monitoring mode. The standard curve was linear ($r^2$ = 0.998) over the concentration range of 1-100 ng/mL. The lower limit of quantification was 1.0 ng/mL using 50 ${\mu}L$ of plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 1.3 to 8.0% and -5.4 to 10.0%, respectively. This method was applied successfully to the pharmacokinetic study of dioscin after oral administration of dioscin at a dose of 29.2 mg/kg in male Sprague-Dawley rats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1552-1563
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• 2016

To examine the cognitive function of onion (Allium cepa L.) beverages (odourless and fortified), we analyzed in vitro neuronal cell protection against $H_2O_2$-induced cytotoxicity and performed in vivo tests on amyloid beta ($A{\beta}$)-induced cognitive dysfunction. Cellular oxidative stress and cell viability were evaluated by DCF-DA assay and MTT assay. These results show that fortified beverage resulted in better neuronal cell protection than odourless beverage at lower concentration ($0{\sim}100{\mu}g/mL$). Fortified beverage also showed more excellent acetylcholinesterase (AChE) inhibitory activity ($IC_{50}$: 4.20 mg/mL) than odourless beverage. The cognitive functions of odourless beverage and fortified beverage in $A{\beta}$-induced neurotoxicity were assessed by Y-maze, passive avoidance, and Morris water maze tests. The results show improved cognitive function in both groups treated with beverages. After in vivo tests, cholinergic activities were determined based on AChE inhibition and acetylcholine levels, and antioxidant activities were measured as SOD, oxidized glutathione (GSH)/total GSH ratio, and MDA levels in mouse brain tissue. In a Q-TOF UPLC/MS system, main compounds were analyzed as follows: odourless beverage (five types of sugars and three types of phenolics) and fortified beverages (six types of phenolics and two types of steroidal saponins).

• The Korean Journal of Pharmacology
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• v.9 no.2
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• pp.39-45
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• 1973

The majority of drugs used in the treatment of superficial fungal infections has limited values due to its low efficacy or development of resistance. For the purpose of searching efficacious agent on the superficial fungal infections induced by dermatophytes which is regarded as the most malicious one, authors examined whether Chinae Rhizoma Extracts have significant on it. Extracts from Smilax china Linne used for the study are water extract (CRWE), ethanol extract (CREE) and methanol extract (CRME). In in vitro studies, the spores of the dermatophytes were inoculated on Sabouraud's glucose agar media which contained three extracts of Chinae Rhizoma in each concentration of $500\;{\mu}g/ml$, $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$ respectively, and also $1,000\;{\mu}g/ml$ of salicylic acid and undecylenic acid $1,000\;{\mu}g/ml$ as comparable drugs. The growth of the dermatophytes were observed for 3 weeks. The species of the dermatophytes used in this experiment were Epidermophyton floccosum, Microsporum canis, Microsporum cookei, Microsporum gypseum, Microsporum nanum, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton tonsurans and Trichophyton verrucosum distributed from The Institute of Tropical Medicine in Belgium. The results of the studies were as follows: 1. The growth of M. canis, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans were slightly inhibited in CRWE $1,000\;{\mu}g/ml$ and CRWE $5,000\;{\mu}g/ml$, and only slight inhibition on the growth of E. floccosum, M. canis and M. gypseum were observed in CRWE $5,000\;{\mu}g/ml$. 2. Complete inhibition of T. rubrum, moderate inhibition of M. nanum & T. tonsurans, and slight inhibition of E. floccosunl, M. canis, M. cookei & T. mentagrophytes in growth were observed in concentration of CREE $500\;{\mu}g/ml$. The growth of M. gypseum was slightly inhibited, moderate inhibition on the growth of M. canis, M. cookei & T. mentagrophytes, and complete inhibition of E. floccosum, M. nanum, T. rubrum & T. tonsurans in growth were observeed by CREE $1,000\;{\mu}g/ml$. With $5,000\;{\mu}g/ml$ of CREE, the growth of E. floccosum, M. canis, M. cookei, M. gypseum, T. mentagrophytes, T. rubrum & T. tonsurans were completely inhibited except T. verrucosum being showed slight inhibition. 3. In CRME $500\;{\mu}g/ml$, slight inhibition of T. verrucosum, moderate inhibition of M. gypseum and complete inhibition of E. floccosum, M. canis, M. cookei, T. mentagrophytes, T. rubrum & T. tonsurans in growth were observed. The growth of E. floccosum, M. canis, M. cookei, M. gypseum, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans were completely inhibited except T. verrucosum being showed moderate inhibition in both CRME $1,000\;{\mu}g/ml$ and CRME $5,000\;{\mu}g/ml$. 4. In $1,000\;{\mu}g/ml$ of undecylenic acid, slight inhibition of T. verrucosum and complete inhibition of E, floccosum, M. canis, M. cookei, M. gypseum, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans in growth were observed. From the above results, it was found that Chinas Rhizoma Alcoholic Extracts(CREE & CRME) exerted significant antifungal activity, and their effects were probably derived from the pharmacological actions of triterpenoidal saponin and steroidal saponin.

• Korean Journal of Plant Resources
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• v.11 no.1
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• pp.51-59
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• 1998

When the contents of the constituents such as total amino acids, free amino acids, volatile organosulfuric compounds and steroidal saponins among three origins in the aerial-and underground parts of Allium victorialis, it was suggested that the characteristic components regarding to quality evaluation could be differed according to the purpose of utilization. For the utilization of amino acids, underground parts of this plant was shown to be better than aerial part. In addition, Ulung island origin was found to contain the highest amino acids content among the three origins though the difference was small. The amino acids showing remarkably high contents were appeared to be arginine, glutamine and asparagine. In the volatile organosulfuric compounds, the origina of Mt. Odae and Mt. Chiri positioned in inland showed higher contents than Ulung island origin geographically positioned in the ocean. Inland origins were shown to contain higher organosulfurie component contents in aerial parts than in underground parts while those of Ulung island origin were higher in underground parts than aerial parts. Underground parts, regarding to saponin constituents, showed higher contents than aerial parts. Underground parts of Ulung island origin were shown to contain more saponins than those of other two origins and the sequence of the contents was in the order of Ulung island>Mt. Chiri>Mt Odac.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.243-250
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• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.

• Journal of Ginseng Research
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• v.43 no.3
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• pp.488-495
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• 2019

Background: Timosaponin AIII (TA3) is a steroidal saponin extracted from Anemarrhena asphodeloides. Here, we investigated the anticancer effects of TA3 in MG63 human osteosarcoma cells. TA3 attenuates migration and invasion of MG63 cells via regulations of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, which are involved with cancer metastasis in various cancer cells. TA3 reduced enzymatic activities and transcriptional expressions of MMP-2 and MMP-9 in MG63 cells. TA3 also inhibited Src, focal adhesion kinase, extracellular signal-regulated kinase (ERK1/2), c-Jun N-terminal kinase (JNK), p38, ${\beta}-catenin$, and cAMP response element binding signaling, which regulate migration and invasion of cells. TA3 induced apoptosis of MG63 cells via regulations of caspase-3, caspase-7, and poly(ADP-ribose) polymerase (PARP). Then, we tested several ginsenosides to be used in combination with TA3 for the synergistic anticancer effects. We found that ginsenosides Rb1 and Rc have synergistic effects on TA3-induced apoptosis in MG63 cells. Methods: We investigated the anticancer effects of TA3 and synergistic effects of various ginseng saponins on TA3-induced apoptosis in MG63 cells. To test antimetastatic effects, we performed wound healing migration assay, Boyden chamber invasion assays, gelatin zymography assay, and Western blot analysis. Annexin V/PI staining apoptosis assay was performed to determine the apoptotic effect of TA3 and ginsenosides. Results: TA3 attenuated migration and invasion of MG63 cells and induced apoptosis of MG63 cells. Ginsenosides Rb1 and Rc showed the synergistic effects on TA3-induced apoptosis in MG63 cells. Conclusions: The results strongly suggest that the combination of TA3 and the two ginsenosides Rb1 and Rc may be a strong candidate for the effective antiosteosarcoma agent.