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.

Bisphenol A (BPA), a known endocrine disruptor, induces toxicity in cells and in experimental animals. Ginseng extracts were evaluated to determine whether they can inhibit BPA-induced toxicity. The antioxidant activity of fresh ginseng extract (WGE), dried white ginseng extract (DGE), and dried red ginseng extract (RGE) was measured using the DPPH assay. WGE and RGE increased DPPH free radical scavenging activity. Cell viability was measured in HepG2 cells following treatment with BPA and ginseng extracts using the MTT assay. DGE and RGE increased HepG2 cell viability following treatment with 200 μM BPA. RGE reduced levels of biochemical markers of liver damage, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) that increased in mice following treatment with BPA. In addition, the regeneration and proliferation of damaged liver cells were significantly increased in RGE-treated mice. Moreover, RGE inhibited hepatic fibrosis in the surrounding area and in the central vein of the liver microstructure. RGE also significantly inhibited BPA-induced cytotoxicity. In addition, RGE protected liver damage and regenerated liver tissues in BPA-treated animals. These results show that RGE may represent a potential candidate drug for the treatment and prevention of liver damage caused by environmental toxins.

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 : Reactive oxygen species (ROS), whether produced endogenously as a consequence of normal cell functions or derived from external sources, pose a constant threat to cells living in an aerobic environment as they can result in severe damage to DNA, protein, and lipids. The effects of Valeriana fauriei extract and fractions on hydrogen peroxide-induced neuronal cell damage are studied. Methods and Results : Oxidative stress plays an important role in the pathological process of neurodegenerative diseases. Valeriana fauriei extract (VFE) and EA fractions (VFEA) was investigated total phenolic contents using method. VFE of total phenolic contents had 2.54 ± 0.01 mg/g, also, VFEA had a 18.78 ± 0.03 mg/g. High phenolic content of the VFEA is expected to better the inhibition of oxidative stress. VFE and VFEA were experimented to inhibit ROS induced 200 μM 3-morpholinosydnonimine (SIN-1). VFE of inhibit SIN-1 induced-ROS dose dependently and signficantly. In addition, VFEA inhibition was also dose dependant and significant. Moreover, Treatment of SH-SY5Y and SK-N-SH cells with VFEA significantly reduced hydrogen peroxide-induced generation of intercellular ROS. Conclusion : From the above results, we may suggest that VFEA might have useful as a material for functional food and pharmaceutics for the pathological process of neurodegenerative diseases.

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 : Ginseng has been commonly used as a traditional oriental medicine for its wide spectrum of medicinal properties, including anti-inflammatory, antitumorigenic, adaptogenic and anti-aging properties. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we have found inflammation-related genes regulated by 20(S)-PPD in mouse bone marrow-derived macrophage (BMDM) to elucidate the role of 20(S)-PPD in inflammatory signaling pathways. Methods and Results : We examined cell viability of BMDM cells after treatment of 20(S)-PPD and found that 20(S)-PPD has no cytotoxicity up to 10 uM. BMDM cells treated with none or 10uM 20(S)-PPD were used for RNA extraction and microarray analysis. It was found that 2 inflammation-related genes are upregulated and 4 genes are downregulated by 20(S)-PPD. Conclusion : These results can give clues to elucidate the role of 20(S)-PPD in inflammatory signaling pathways.

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.

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 was carried out to evaluate the preventive effect of three forms of Korean ginseng roots (fresh, white and red) against bisphenol A (BPA) toxicity in mouse male germ cells (GC-2spd, TM3, TM4). ROS (reactive oxygen species) generation were measured by DCF-DA (2’,7’-dichlorohydrofluorescein diacetate) assay. Also, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression levels of apoptosis- related genes, Bax (pro-apoptotic gene) and Bcl2 (anti-apoptotic gene). ROS generation was increased by 50 μM BPA, but definitely decreased by treatment with Korean ginseng extracts (fresh, white and red) in mouse male germ cells. In especial, Korean fresh ginseng extract reduced significantly ROS production to normal control. In addition, Korean fresh and white ginseng extracts suppressed the apoptosis of mouse male germ cells by fine-tuning mRNA levels of apoptotic genes changed by BPA. In general, Korean fresh ginseng extract was more effective than white ginseng extract for reducing BPAinduced oxidative stress and apoptosis in mouse male germ cells. Therefore, Korean fresh and white ginseng may help to alleviate biphenol A toxicity in mouse male germ cells.