A synthetic chalcone derivative, 2-hydroxy-3',5,5'-trimethoxychalcone (DK-139), suppresses the TNFα-induced invasive capability of MDA-MB-231 human breast cancer cells by inhibiting NF-κB-mediated GROα expression.

2-Hydroxy-3',5,5'-trimenthoxyochalcone (DK-139) is a synthetic chalcone-derived compound. This study evaluated the biological activity of DK-139 on the inhibition of tumor metastasis. Growth-regulated oncogene-alpha (GROα) plays an important role in the progression of tumor development by stimulating angiogenesis and metastasis. In this study, DK-139 inhibited tumor necrosis factor alpha (TNFα)-induced GROα gene promoter activity by inhibiting of IκB kinase (IKK) in MDA-MB231 cells. In addition, DK-139 prevented the TNFα-induced cell migration, F-actin formation, and invasive capability of MDA-MB-231 cells. These findings suggest that DK-139 is a potential drug candidate for the inhibition of tumor cell locomotion and invasion via the suppression of NF-κB-mediated GROα expression.

UPLC-MS/MS-Based Profiling of Eicosanoids in RAW264.7 Cells Treated with Lipopolysaccharide.

While both the pro- and anti-inflammatory effects of several eicosanoids have been widely studied, the degree of inflammation in cells that results from various eicosanoids has yet to be comprehensively studied. The objective of this study was to assess the effect of lipopolysaccharide (LPS) treatment on eicosanoid content in RAW264.7 cells. An Ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS)-based profiling method was used to analyze the eicosanoid contents of RAW264.7 cells treated with different LPS concentrations. The profiling data were subjected to statistical analyses, such as principal component analysis (PCA) and hierarchical clustering analysis. LPS treatment increased nitric oxide production and secretion of pro-inflammatory cytokines, such as tumor necrosis factor-α and interleukin-6, in a concentration-dependent manner. In total, 79 eicosanoids were identified in the cells. RAW264.7 cells treated with different LPS concentrations were well differentiated in the PCA score plot. A heatmap was used to identify the eicosanoids that were up- or down-regulated according to the degree of inflammation and LPS concentration. Thirty-nine eicosanoids were upregulated and seven were down-regulated by LPS treatment in a concentration-dependent manner. Our novel UPLC-MS/MS technique can profile eicosanoids, and can evaluate the correlations between inflammation and eicosanoid metabolism.

Global Profiling of Various Metabolites in Platycodon grandiflorum by UPLC-QTOF/MS.

In this study, a method of metabolite profiling based on UPLC-QTOF/MS was developed to analyze Platycodon grandiflorum. In the optimal UPLC, various metabolites, including major platycosides, were separated well in 15 min. The metabolite extraction protocols were also optimized by selecting a solvent for use in the study, the ratio of solvent to sample and sonication time. This method was used to profile two different parts of P. grandiflorum, i.e., the roots of P. grandiflorum (PR) and the stems and leaves of P. grandiflorum (PS), in the positive and negative ion modes. As a result, PR and PS showed qualitatively and quantitatively different metabolite profiles. Furthermore, their metabolite compositions differed according to individual plant samples. These results indicate that the UPLC-QTOF/MS-based profiling method is a good tool to analyze various metabolites in P. grandiflorum. This metabolomics approach can also be applied to evaluate the overall quality of P. grandiflorum, as well as to discriminate the cultivars for the medicinal plant industry.

Induction of human microsomal prostaglandin E synthase 1 by activated oncogene RhoA GTPase in A549 human epithelial cancer cells.

Oncogenic RhoA GTPase has been investigated as a mediator of pro-inflammatory responses and aggressive carcinogenesis. Among the various targets of RhoA-linked signals, pro-inflammatory prostaglandin E(2) (PGE(2)), a major prostaglandin metabolite, was assessed in epithelial cancer cells. RhoA activation increased PGE(2) levels and gene expression of the rate-limiting PGE(2) producing enzymes, cyclooxygenase-2 and microsomal prostaglandin E synthase 1 (mPGES-1). In particular, human mPGES-1 was induced by RhoA via transcriptional activation in control and interleukin (IL)-1ß-activated cancer cells. To address the involvement of potent signaling pathways in RhoA-activated mPGES-1 induction, various signaling inhibitors were screened for their effects on mPGES-1 promoter activity. RhoA activation enhanced basal and IL-1ß-mediated phosphorylated nuclear factor-κB and extracellular signal-regulated kinase1/2 proteins, all of which were positively involved in RhoA-induced gene expression of mPGES-1. As one potent down-stream transcription factor of ERK1/2 signals, early growth response gene 1 product also mediated RhoA-induced gene expression of mPGES-1 by enhancing transcriptional activity. Since oncogene-triggered PGE(2) production is a critical modulator of epithelial tumor cells, RhoA-associated mPGES-1 represents a promising chemo-preventive or therapeutic target for epithelial inflammation and its associated cancers.

Isolation and identification of phytochemicals and biological activities of Hericium ernaceus and their contents in Hericium strains using HPLC/UV analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Hericium ernaceus has been traditionally used for the treatment of dyspepsia, gastric ulcer and enervation in traditional Chinese medicine for a long time. AIM OF THE STUDY: To examine the effect of Hericium strains on their ability to inhibit LPS and interferon-γ induced NO production in cell culture and the bioassay correlation of hericenone C, D, F, isolated from H. ernaceus. MATERIALS AND METHODS: Hericenone C, D, F were isolated from H. ernaceus by open column chromatography and identified on the basis of spectroscopic analyses including (1)H NMR, (13)C NMR and MS. The amounts of hericenone C, D, and F in Hericium strains were determined by HPLC/UV analysis. In order to investigate the anti-inflammatory effect of Hericium strains extracts, RAW 264.7 cells were treated with 200µg/mL of Hericium strains extracts for 48h. Cell growth was assessed by MTT assay. RESULTS: Phytochemical constituents were isolated from H. ernaceus by open column chromatography. Their structures were elucidated as hericenones C, D, and F on the basis of spectroscopic analyses including (1)H NMR, (13)C NMR and MS. The amounts of hericenones C, D, and F in Hericium strains were determined by HPLC/UV analysis. Hericenones C, D, and F contents were highest in Norugungdenglee-2 (8.289±0.593mg/g), KFRI-1453 (4.657±0.462mg/g), and KFRI-1093 (5.408±0.420mg/g) strains, respectively. All Hericium strains extracts tested inhibited the lipopolysaccharide- and interferon-γ-induced inflammatory activity of RAW264.7 cells. The strain KFRI-1093 about 39.6% reduced NO generation with compared to control. CONCLUSION: We believe that the anti-inflammatory effect of KFRI-1093 was due to hericenone F content. Our results contribute towards validation of the traditional use, natural drugs and health supplements. And also, the developed simple, accurate and rapid LC method can be used determinate the content of hericenones from other Hericium strains.

Hepatoprotective Effect and Synergism of Bisdemethoycurcumin against MCD Diet-Induced Nonalcoholic Fatty Liver Disease in Mice.

Nonalcoholic fatty liver disease (NAFLD), the hepatic manifestation of the metabolic syndrome, has become one of the most common causes of chronic liver disease over the last decade in developed countries. NAFLD includes a spectrum of pathological hepatic changes, such as steatosis, steatohepatitis, advanced fibrosis, and cirrhosis. Bisdemethoxycurcumin (BDMC) is polyphenolic compounds with a diarylheptanoid skeleton, curcumin close analogues, which is derived from the Curcumae Longae Rhizoma. While the rich bioavailability research of curcumin, BDMC is the poor studies. We investigated whether BDMC has the hepatoprotective effect and combinatory preventive effect with silymarin on methionine choline deficient (MCD)-diet-induced NAFLD in C57BL/6J mice. C57BL/6J mice were divided into five groups of normal (normal diet without any treatment), MCD diet (MCD diet only), MCD + silymarin (SIL) 100 mg/kg group, MCD + BDMC 100 mg/kg group, MCD + SIL 50 mg/kg + BDMC 50 mg/kg group. Body weight, liver weight, liver function tests, histological changes were assessed and quantitative real-time polymerase chain reaction and Western blot analyses were conducted after 4 weeks. Mice lost body weight on the MCD-diet, but BDMC did not lose less than the MCD-diet group. Liver weights decreased from BDMC, but they increased significantly in the MCD-diet groups. All liver function test values decreased from the MCD-diet, whereas those from the BDMC increased significantly. The MCD- diet induced severe hepatic fatty accumulation, but the fatty change was reduced in the BDMC. The BDMC showed an inhibitory effect on liver lipogenesis by reducing associated gene expression caused by the MCD-diet. In all experiments, the combinations of BDMC with SIL had a synergistic effect against MCD-diet models. In conclusion, our findings indicate that BDMC has a potential suppressive effect on NAFLD. Therefore, our data suggest that BDMC may act as a novel and potent therapeutic agent against NAFLD.

Ethanol extract of Lithospermum erythrorhizon Sieb. et Zucc. promotes osteoblastogenesis through the regulation of Runx2 and Osterix.

Bone remodeling and homeostasis are largely the result of the coordinated action of osteoblasts and osteoclasts. Osteoblasts are responsible for bone formation. The differentiation of osteoblasts is regulated by the transcription factors, Runx2 and Osterix. Natural products of plant origin are still a major part of traditional medicinal systems in Korea. The root of Lithospermum erythrorhizon Sieb. et Zucc. (LR), the purple gromwell, is an herbal medicine used for inflammatory and infectious diseases. LR is an anti-inflammatory and exerts anticancer effects by inducing the apoptosis of cancer cells. However, the precise molecular signaling mechanisms of osteoblastogenesis as regards LR and osteoblast transcription are not yet known. In this study, we investigated the effects of ethanol (EtOH) extract of LR (LES) on the osteoblast differentiation of C2C12 myoblasts induced by bone morphogenetic protein 4 (BMP4) and the potential involvement of Runx2 and Osterix in these effects. We found that the LES exhibited an ability to induce osteoblast differentiation. LES increased the expression of the osteoblast marker, alkaline phosphatase (ALP), as well as its activity, as shown by ALP staining and ALP activity assay. LES also increased mineralization, as shown by Alizarin Red S staining. Treatment with LES increased the protein levels (as shown by immunoblotting), as well as the transcriptional activity of Runx2 and Osterix and enhanced osteogenic activity. These results suggest that LES modulates osteoblast differentiation at least in part through Runx2 and Osterix.

Melanin Biosynthesis Inhibition Effects of Ginsenoside Rb2 Isolated from Panax ginseng Berry.

Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng. Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, (1)H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 µM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.

Identification of genes possibly related to storage root induction in sweet potato.

To identify genes related to initiation of storage root development in sweet potato, a cDNA library was constructed with early stage storage roots (0.3-1 cm in diameter). Single-pass sequences of the 5' ends of 2859 sweet potato cDNA clones were assembled into 483 clusters and 442 singletons. Comparison of sweet potato expressed sequence tags (ESTs) to nodulation/tumorigenesis-related sequence databases (nodule-, tumor-, potato tuber- and development-related sequences) revealed that homologs of 39 sweet potato EST sequences potentially involved in gene regulation, signal transduction and development were present in at least one of the nodulation/tumorigenesis-related sequence databases. Northern blot analyses of these 39 sequences identified 22 differentially expressed genes in early stage storage root and fibrous root. These differentially expressed genes will be potential candidates for research to elucidate the molecular processes related to sweet potato storage root induction.

Postharvest strategies for deoxynivalenol and zearalenone reduction in stored adlay (Coix lachryma-jobi L.) grains.

Improperly practiced postharvest procedures can pose mycotoxin-related risks in the production of medicinal herbs. As a health food with pharmacological supplements, cereal-based adlay has been broadly used in oriental medical practice. Compared with the standard production protocol, three provisional critical control points (CCPs) in the conventional procedure were identified and assessed for mycotoxin contamination in the adlay from small farms in Korea. Although various mycotoxins were present, the prevalence of deoxynivalenol (DON) or zearalenone (ZEN) was relatively high in the adlay. In terms of drying conditions, field drying in the conventional pathway was associated with more exposure to DON than heated-air drying. Moreover, the DON or ZEN levels in chaff were higher than the levels in the inner grain, suggesting that the hulling process as another CCP would reduce the DON or ZEN exposure. In particular, the DON or ZEN levels in adlay stored for protracted periods without dehulling were very high, but a lower storage temperature of 12°C was not effective at significantly reducing these mycotoxins. In this case, the inner grain was more contaminated with DON or ZEN than the chaff after protracted storage because surface fungi, which produce mycotoxins, can penetrate deep into grain with time. Heated-air drying and nonprotracted storage limited DON contamination in adlay. More importantly, an early dehulling process should be adopted as an easy preventive action to reduce the risk of exposure to DON or ZEN in adlay postharvest. This is monitored as a central CCP for safer production of adlay from local farms.

Combination Therapy of Sophoraflavanone B against MRSA: In Vitro Synergy Testing.

Sophoraflavanone B (SPF-B), a known prenylated flavonoid, was isolated from the roots of Desmodium caudatum. The aim of this study was to determine the antimicrobial synergism of SPF-B combined with antibiotics against methicillin-resistant Staphylococcus aureus (MRSA). MRSA, a multidrug-resistant pathogen, causes both hospital- and community-acquired infections worldwide. The antimicrobial activity of SPF-B was assessed by the broth microdilution method, checkerboard dilution test, and time-kill curve assay. The MIC of SPF-B for 7 strains of S. aureus ranges from 15.6 to 31.25 µ g/mL determined. In the checkerboard method, the combinations of SPF-B with antibiotics had a synergistic effect; SPF-B markedly reduced the MICs of the ß -lactam antibiotics: ampicillin (AMP) and oxacillin (OXI); aminoglycosides gentamicin (GET); quinolones ciprofloxacin (CIP) and norfloxacin (NOR) against MRSA. The time-kill curves assay showed that a combined SPF-B and selected antibiotics treatment reduced the bacterial counts below the lowest detectable limit after 24 h. These data suggest that the antibacterial activity of SPF-B against MRSA can be effectively increased through its combination with three groups of antibiotics ( ß -lactams, aminoglycosides, and quinolones). Our research can be a valuable and significant source for the development of a new antibacterial drug with low MRSA resistance.