Background : While the anti-inflammatory effects of 20 (S)-ginsenoside Rg3 (Rg3) have been studied, it remains unclear how Rg3 regulates lipid metabolism in inflammatory macrophages. Thus, in this study, we characterized some eicosanoids related to the anti-inflammatory effects of 20 (S)-ginsenoside Rg3 in murine macrophages. Methods and Results : UPLC-MS/MS was used to profile various eicosanoids from RAW264.7 cells treated with lipopolysaccharide (LPS) and Rg3. The profiling data were statistically analyzed by principal component analysis, hierarchical clustering analysis and analysis of variance. The anti-inflammatory effect of Rg3 was validated by assessing the levels of nitric oxide, tumor necrosis factor-α, and interleukin-6 in the activated macrophages treated with Rg3. A total of 69 eicosanoids were analyzed in RAW264.7 cells. Principal component and hierarchical cluster analyses differentiated control cells from cells treated with LPS, Rg3, or LPS + Rg3 for 12 or 24 h. Furthermore, some differentially regulated compounds were found between macrophages treated with LPS for 24 h and those treated with LPS + Rg3 for 24 h. Conclusion : Rg3 alters eicosanoid metabolism in activated macrophages treated with LPS. Furthermore, we identified several eicosanoids correlated with the anti-inflammatory activity of Rg3.

Background : Methicillin-Resistant Staphylococcus aureus (MRSA) is a multidrug-resistant (MDR) strain. Especially, MRSA is developing resistance to available antibacterial agents and causing complications in the treatment of infections related to skin, soft tissue, respiratory, bone, joint, and endovascular disorders. Therefore, antibacterial agent combination therapy appears to be a useful option, particularly in developing countries where antibiotic availability is limited. (+)-Usnic acid (UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. Methods and Results : In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against MRSA. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid (EDTA) was used. In the other hand, Sodium azide (NaN3) was used as inhibitors of ATPase. These results suggest that the antibacterial effect of UA was potentiated by membrane-binding agents and ABC transporter-inhibiting agents, implying that antibacterial activity is associated with damage of the cell wall and inhibition of ATPase function by UA. Conclusion : UA and in combination with EDTA and NaN3 could lead to the development of new combination antibiotics against MRSA infection. The results of this study appear to be promising, and they are expected to enhance the use of natural products as drugs.

Background: This study was performed to evaluate the protective effect of Saururus chinensis ethanol extract (SCE) against styrene toxicity in mouse spermatocyte cells [GC-2spd (ts) cell line]. Methods and Results: Cytotoxicity in mouse spermatocyte cells was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Generation of reactive oxygen species (ROS) was determined using 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to quantify the mRNA and protein expression levels, resepectiviely, of stress or apoptosis-related genes including p21, p53, heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), Bax, Bcl-2, and caspase-3. The results of the MTT assay showed that 50 ㎍/㎖ SCE did not affect cell viability. ROS generation in mouse spermatocyte cells increased by treatment with 100 μM styrene, and decreased by co-treatment with SCE. SCE repressed the mRNA expression of stress-related genes, which increased by styrene treatment. In addition, SCE inhibited the apoptosis of mouse spermatocyte cells by ameliorating mRNA and protein levels of apoptotic genes that were altered by styrene treatment. Conclusions: These results suggest that SCE may alleviate styrene toxicity in mouse spermatocyte cells by reducing ROS stress and regulating genes related to styrene toxicity.

Background: The study was conducted to elucidate the extraction conditions under which white ginseng has cognition-improving efficacy.Methods and Results: Extracts from white ginseng under different solvent and temperature conditions were analyzed for ginsenoside content and inhibitory effect on N-methyl-D-aspartate (NMDA) receptor and acetylcholinesterase. The total ginsenoside contents and amounts of ginsenoside Rb1 plus ginsenoside Rg1 from the 1st extracts (prepared with EtOH/H2O as solvent) were higher than those from the 2nd extracts (extracted with H2O after the 1st EtOH/H2O extraction). The contents in the 1st and 2nd extracts produced at 80°C were also higher than those obtained at 50°C. Samples from the 1st extraction at 80°C indicated higher inhibitory activities on NMDA receptors-whose excessive activation is thought to mediate the calcium-dependent neurotoxicity associated with several neurodegenerative diseases-than those from the 2nd extraction. Among the samples prepared at varying temperatures, the extract prepared at 50°C showed the highest suppression activity on NMDA receptors. Note, however, that the extracts from the 2nd extraction at 50°C inhibited acetylcholinesterase-whose inhibition could be a therapeutic strategy for neurodegenerative diseases with cognitive deficits and memory malfunction-more effectively than those from the 1st extraction.Conclusions: To enhance the cognition-improving activity of white ginseng extract, it is suggested that the extracts be utilized after being combined the 1st extracts (made with EtOH/H2O solvent) and the 2nd extracts (prepared with H2O) at low temperature.

Background : Acanthopanax sessiliflorus (Rupr. et Maxim) Seem, belonging to the Araliaceae family, is widely distributed in Korea, China, and Japan. The plants belonging to Acanthopanax species are traditionally used in Korea as anti-rheumatoid arthritis, anti-inflammatory and anti-diabetic drugs and are recognized to have ginseng-like activities. A simple and sensitive high-performance liquid chromatographic (HPLC) method was developed and validated for independent analysis of major compounds and chlorogenic acid in A. sessiliflorus fruits. Chlorogenic acid was reported that prevent cancer and cardiovascular disease in vivo. Also, it has antioxidant effect in vitro test. In the previous experiment, chlorogenic acid were found in A. sessiliflorus fruits. This study was performed to identification of the major compounds and investigate the method validation for the determination of chlorogenic acid in A. sessiliflorus fruits. Methods and Results : Three major compounds were recorded on a Varian Unity Inova AS-400 FT-NMR spectrometer and analyzed by the new HPLC analysis method. HPLC analysis was carried out using an Waters e2695 and PDA detector. The new analyasis method was validated by the measurement of intra-day, inter-day precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) of chlorogenic acid. The results showed that the correlation coefficient (R2) for the calibration curves of chlorogenic acid was 0.997 in terms of linearity. The limit of detection (LOD) and limit of quantification (LOQ) were 0.565 ㎍/ml and 2.88 ㎍/ml, respectively. There was no interfering peak observed each other and HPLC system was suitable for analysis showing goodness of peak and high precision. Conclusion : This method is suitable to detect and quantify major compounds in A. sessiliflorus fruits. Furthermore, the result will be applied to establish chlorogenic acid as an standard compound for A. sessiliflorus fruits.

Background : Allergy is a common disease caused by type I hypersensitivity reaction of the immune system. Unfortunately, there is no proper treatment for allergy. Therefore, the discovery of therapeutic drugs for allergy is essential. In this study, the crude extracts of 56 plant parts were screened for anti-allergy effects in RBL-2H3 cells. Methods and Results : IgE-sensitized RBL-2H3 cells were individually treated with 56 extracts of medicinal herbs at the final concentration of 20 ㎍/㎖ and stimulated with the antigen DNP-BSA. β-Hexosaminidase release, interleukin-4 (IL-4) secretion, and cell viability in the sample treated cells were measured. Among the tested samples, extracts from the root of Polygonatum stenophyllum Maxim., aerial part of Acer triflorum Kom., and leaf of Pyrus pyrifolia var. culta (Makino) Nakai showed inhibitory effects on β-hexosaminidase release. The aerial part of Peucedanum japonicum Thunberg and seed of Panax ginseng C.A.Mey. showed suppressive activities on IL-4 secretion. All of the extracts were not cytotoxic at the tested concentration. Conclusion : From the result, six extracts including Polygonatum stenophyllum Maxim. (root) and Acer triflorum Kom. (aerial part) inhibited both β-hexosaminidase release and IL-4 secretion in IgE-sensitized RBL-2H3 cells. The use of these extracts for developing anti-allergy materials is suggested.

Background : Five medicinal herbs have been selected from the preliminary screening for in vitro anti-allergy activity (in RBL-2H3 cells). The present study is conducted to investigate the inhibitory effects of the medicinal herbs on allergic inflammation in other kind of cells. Methods and Results : Cells treated with five extracts prepared from Betula costata Trautv. (aerial part), Camellia sinensis L. (aerial part), Polygonatum stenophyllum Maxim. (root), Pyrus pyrifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. (root) were measured for mRNA levels of TNF-α on HaCaT keratinocytes stimulated by TNF-α /INF-γ and for mRNA levels of IL-2 in Jurkat T cells mediated by PMA/A23187. Pre-treatment with the five extracts reduced the mRNA levels of TNF-α in HaCaT cells and mRNA levels of IL-2 in Jurkat T cells. In particular, the leaf extract of Pyrus pyrifolia var. culta (Makino) Nakai significantly and dose-dependently decreased the mRNA levels of TNF-α and IL-2. To determine the toxicity of the extracts from the selected medicinal herbs to HaCaT cells and Jurkat T cells, the viabilities of the cells treated with several concentrations of the five extracts were measured by MTT assay. Extracts of Polygonatum stenophyllum Maxim. (root), Pyrus pyrifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. (root) (up to 250 ㎍/㎖) did not show cytotoxic effects on HaCaT cells and Jurkat T cells. On the other hand, 250 ㎍/㎖ of extracts of Betula costata Trautv. (aerial part) and Camellia sinensis L. (aerial part) reduced cell viability in both cells. Conclusion : These results demonstrate that the leaf extract of Pyrus pyrifolia var. culta (Makino) Nakai has an anti-inflammatory effect by inhibiting pro-inflammatory cytokine expression. Therefore, the leaf of Pyrus pyrifolia var. culta (Makino) Nakai can be a useful resource for the development of anti-allergy/anti-inflammatory materials.

Background : Ginseng, one of most famous traditional oriental medicines, has been known for a number of pharmacological properties including anti-tumor, anti-diabetic, anti-fatigue, anti-stress, anti-oxidative, and anti-aging effects. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we investigated the synergistic anticancer effect of 20(S)-PPD and temozolomide (TMZ) and the mechanism of 20(S)-PPD on glioblastoma cells. Methods and Results : We examined cell viability and the morphological changes of C6 cells after treatment of 20(S)-PPD and TMZ. 20(S)-PPD showed a potent antiproliferative activity against C6 cells by triggering apoptosis. 20(S)-PPD-induced apoptosis was characterized by a dose-dependent mitochondrial damage. 20(S)-PPD and TMZ had a synergistic effect in increasing mitochondrial damage via caspase 3 activation. Conclusion : These results revealed an unexpected mechanism of 20(S)-PPD and TMZ, triggering a mitochondrial-mediated apoptosis in C6 cells. Our findings encourage further studies of 20(S)-PPD as a promising chemopreventive agent against glioblastoma.

Background : Recently, ginseng (Panax ginseng C.A. Meyer.) berry has been used as a health-promoting supplements. Also, Mulberries (Morus alba L.) fruit have been used in traditional herbal medicine to treat and prevent diabetes. In this study, we measured the cytotoxicity after fermentation of the extracts in Panax Ginseng Berry and Mulberry Fruit. Methods and Results : The extracts were prepared by decoction for 3 hours in distilled water (100 g/L). The dried extract was then dissolved in phosphate-buffered saline (PBS) in preparation for use. Cell viability was examined by an MTT assay. RAW 264.7 cells were seeded at 1 × 104/mL densities in 96-well plates. Each grouping had a non-treated group as the control. The extracts were added to each well and incubated for 24 hours at 37°C and 5% CO2. The MTT solutions (5 mg/mL) were added to each well, and the cells were cultured for another 2 hours. The supernatant was then discarded, and 100 μL of dimethyl sulfoxide was added to each well. The optical density was read at 540 nm. Conclusion : Probiotics and prebiotics modulate the composition of human and domestic animal gut microbiota. The beneficial effects may result from suppression of harmful microorganisms or stimulation of organisms which contribute in a positive way to the nutrition and health of human and domestic animal. Recently, fermentation using microorganisms for the production of more effective compounds has been extensively studied. In particular, the novel pharmacological effects of a new compound generated by fermentation have been reported. Some previous studies have demonstrated that Fermented herbal medicine extract showed better bioactivity than normal herbal Plants extract when used at the same concentration.

A new extraction method-heated ultrasonic extraction was qualitatively and quantitatively analyzed for the extraction of major ginsenosides from ginseng extract; this new high-performance liquid chromatography (HPLC) method was compared with the official extraction method of Korean industrial standards and standard for health functional food. Methods and Results : Ginsenoside compounds were analyzed for 35 minutes by the new HPLC analysis method using a Halo® RP-Amide column. The new HPLC analysis method was validated by the measurement of intra-day and inter-day precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of each ginsenoside. The correlation coefficients (r2) for the calibration curves of the ginsenoside compounds were over 0.9997 in terms of linearity. The heated ultrasonic extraction method using ultrasonication for 30 minutes at 50℃ yielded higher amount of ginsenosides than the extraction method of the Korean industrial standards owing to the enhancement of extraction efficiency. Conclusions : Compared to the other extraction methods, the heated ultrasonic extraction method yielded a higher amount of ginsenoside Rb1 than Rg1 index compounds for the quality evaluation of ginseng roots.

This study has been conducted to establish the optimal extraction process and HPLC analysis method for thedetermination of marker compounds as a part of the materials standardization for the development of health functionalfood materials from Astragali radix. Five extraction conditions including the shaking extraction at room temperature andthe reflux extraction at 85℃ with 30%, 50% and 95% ethanol were evaluated. Reflux extraction with 50% ethanol showedthe highest extraction yield as 27.27±2.27%, while the extraction under reflux with 95% ethanol showed significantly thelowest yield of 10.55±0.24%. The quantitative determination methods of calycosin-7-O-β-D-glucoside and calycosin asmarker compounds of Astragali radix extracts were optimized by HPLC analysis using a Thermo Hypersil column(4.6×250㎜, 5㎛) with the gradient elution of water and acetonitrile as the mobile phase at the flow rate of 0.8mL min-¹and a detection wavelength of 230㎚. The HPLC/UV method was applied successfully to the quantification of two markercompounds in Astragali radix extracts after validation of the method with the linearity, accuracy and precision. The con-tents of calycosin-7-O-β-D-glucoside and calycosin in 50% ethanol extracts by reflux extraction were significantly higher as1,700.3±30.4 and 443.6±8.4㎍ g-1, respectively, comparing with those in other extracts. The results indicate that thereflux extraction with 50% ethanol at 85℃ is optimal for the extraction of Astragali radix, and the established HPLCmethod are very useful for the evaluation of marker compounds in Astragali radix extracts to develop the health functionalmaterial from Astragali radix.

This study investigates the effect of supercritical fluid extract (CMPB803-C) of Lithospermum erythrorhizon,shikonin and acetylshikonin isolated from Lithospermum erythrorhizon on IL-1β-induced chondrocytes and monosodiumiodoacetate (MIA)-induced osteoarthritis in rat. Shikonin (50μM) and acetylshikonin (3μM) treatment reduced signifi-cantly the mRNA expression and enzyme activity of matrix metalloproteinase (MMP)-1, −3 and −13 in IL-1β-inducedSW1353 chondrosarcoma cells. The chondro-protective effects of CMPB803-C and acetylshikonin were than analyzed in arat OA model using a single intra-articular injection of MIA (1㎎) in the right knee joint. CMPB803-C (200㎎/㎏) or ace-tylshikonin (5㎎/㎏) was orally administered daily for two weeks starting after 1 week of MIA injection. In the histologicalobservation, CMPB803-C and acetylshikonin clearly improved OA lesions being comparable to or better that control group.Our results demonstrated that CMPB803-C and acetylshikonin as active compound of Lithospermum erythrorhizon have astrong chondro-protective effect in OA rats, which likely attributes to its anti-inflammatory activity and inhibition ofMMPs production.