A hot-water extract of Sanguisorba officinalis ameliorates endotoxin-induced septic shock by inhibiting inflammasome activation.

The inflammasome is a multiprotein signaling complex that mediates inflammatory innate immune responses through caspase 1 activation and subsequent IL-1ß secretion. However, because its aberrant activation often leads to inflammatory diseases, targeting the inflammasome holds promise for the treatment of inflammation-related diseases. In this study, it was found that a hot-water extract of Sanguisorba officinalis (HSO) suppresses inflammasome activation triggered by adenosine 5'-triphosphate, nigericin, microbial pathogens, and double stranded DNA in bone marrow-derived macrophages. HSO was found to significantly suppress IL-1ß production in a dose-dependent manner; this effect correlated well with small amounts of caspase 1 and little ASC pyroptosome formation in HSO-treated cells. The anti-inflammatory activity of HSO was further confirmed in a mouse model of endotoxin-induced septic shock. Oral administration of HSO reduced IL-1ß titers in the serum and peritoneal cavity, increasing the survival rate. Taken together, our results suggest that HSO is an inhibits inflammasome activation through nucleotide-binding domain and leucine-rich repeat pyrin domain 3, nucleotide-binding domain and leucine-rich repeat caspase recruitment domain 4 and absent in melanoma 2 pathways, and may be useful for treatment of inflammasome-mediated diseases.

Development and Method Validation of Analysis of Urushiol in Sumac and Food Ingredients in Korea.

This study developed an analytical method to determine the urushiol content in sap and several foods. The full process for urushiol analysis consists of extraction, trimethylsilyl silylation, analysis, and identification via GC-MS, with each step optimized to attain the required accuracy and precision. Urushiol was separated from sap via liquid-liquid extraction and was derivatized via silylation. The components were analyzed using a polar capillary column and identified using GC-MS. The deviations of relative retention times and retention time windows were within 0.001 and 0.02 min, which satisfied the criteria of 0.06 and 0.03 min, respectively. The response of the urushiol standards tested was found to be linear in the investigated concentration range, with a correlation coefficient of 0.998. The LODs were between 1.74 and 2.67 µg/mL.

A hot water extract of Aralia cordata activates bone marrow-derived macrophages via a myeloid differentiation protein 88-dependent pathway and protects mice from bacterial infection.

In traditional Asian medicine, Aralia cordata (AC) is a known as a pain reliever and anti-inflammatory drug. Although several of its biological activities have been reported, the immunomodulatory effects of a hot water extract of AC (HAC) have not yet been described. The aim of this study was to investigate whether HAC modulates the activation of macrophages, which play important roles in innate immune responses against microbial pathogens, and if so, to determine the molecular mechanisms by which HAC mediates this process. It was found that HAC activates bone marrow-derived macrophages (BMDM) and increases amounts of nitric oxide and proinflammatory cytokines in a dose-dependent manner. In addition, HAC was found to induce phosphorylation of NF-κB and mitogen-activated protein kinases (MAPKs), including c-Jun N-terminal kinases, extracellular signal-regulated kinases and p38. Interestingly, these effects were absent in BMDM prepared from myeloid differentiation protein 88-knockout mice. Polysaccharides from HAC exerted stronger immunostimulatory effects than HAC itself. Furthermore, orally administered HAC clearly enhanced clearance of the intracellular pathogen Listeria monocytogenes by boosting innate immune responses. These results demonstrate that HAC exerts immunostimulatory effects through the TLR/MyD88 and NF-κB/MAPK signal transduction pathways.

Differential Modulation of Lipopolysaccharide-Induced Inflammatory Cytokine Production by and Antioxidant Activity of Fomentariol in RAW264.7 Cells.

Medicinal mushrooms have been used worldwide to treat cancer and modulate the immune system. Over the last several years, there has been increasing interest in isolating bioactive compounds from medicinal mushrooms and evaluating their health beneficial effects. Fomes fomentarius is used in traditional oriental medicine and is known to possess antioxidant, anti-inflammatory, antidiabetic, and antitumor effects. In the present study, we isolated fomentariol from Fomes fomentarius and investigated its anti-inflammatory effect in murine macrophages (RAW264.7 cells) stimulated with lipopolysaccharides. Fomentariol inhibited the production of nitric oxide and intracellular reactive oxygen species triggered by lipopolysaccharides. Interestingly, fomentariol differentially regulated cytokine production triggered by lipopolysaccharides. Fomentariol effectively suppressed the production of interleukin-1ß and interleukin-6 but not tumor necrosis factor-α. The inhibitory effect of fomentariol against nitric oxide, interleukin-1ß, and interleukin-6 production was possibly mediated by downregulation of the extracellular signal-regulated kinase signaling pathway. Taken together, our results suggest that fomentariol differentially modulated inflammatory responses triggered by lipopolysaccharides in macrophages and is one of the bioactive compounds that mediate the physiological effects of Fomes fomentarius.

Rapid method for determination of anthocyanin glucosides and free delphinidin in grapes using u-HPLC.

A rapid method for the determination of anthocyanin glucosides and free delphinidin in food using ultra-high-performance liquid chromatography (u-HPLC) was validated through tests of precision, accuracy and linearity. The u-HPLC separation was performed on a reversed-phase C18 column (particle size 2 µm, i.d. 2 mm, length 100 mm) with a photodiode array detector. The limits of detection and quantification of the u-HPLC analyses ranged from 0.07 to 0.65 mg/kg for nine types of anthocyanin glucosides and from 0.08 to 0.26 mg/kg for delphinidin aglycone. The intra-day and inter-day precision of individual anthocyanin glucosides and delphinidin aglycone were less than 9.42%. All calibration curves showed good linearity (coefficient of determination = 0.99) within the tested ranges. The rapid and simultaneous u-HPLC method presented in this study significantly improved the speed, sensitivity and resolution of the analyses of nine types of anthocyanin glucosides and free delphidinin aglycone in grapes.

Determination of 22 ginsenosides in ginseng products using ultra-high-performance liquid chromatography.

A reverse-phase ultra-high-performance liquid chromatography (u-HPLC) method was developed for the rapid quantification of 22 ginsenosides in ginseng products. The proposed method for the analysis of ginsenosides is based on a heating-block method without further treatment. The u-HPLC separation was performed on a reversed C18 column (100 × 2 mm id, particle size 2 µm) followed by ultraviolet detection at 203 nm. Aqueous 50% methanol was used as the extraction solvent. The optimum amount of extraction solvent and the optimum extraction time were 20 mL and 20 min (extracted twice with 10 mL), respectively. The method validation parameters yielded good results for linearity, precision, accuracy and recovery. The recovery of ginsenosides from ginseng powder was greater than 98.1% and the limits of detection and quantification of the u-HPLC analysis were >0.6 and >1.8 mg/kg for ginsenosides. The calibration graphs for ginsenosides were linear from approximately 2.6 to 40.4 mg/kg for u-HPLC. The inter-day and intra-day precisions (relative standard deviation values) were <14.6 and 14.7%, except for Rg2(R) + Rh1(R).

Determination of hexanal as an oxidative marker in vegetable oils using an automated dynamic headspace sampler coupled to a gas chromatograph/mass spectrometer.

An automated dynamic headspace sampler coupled to a gas chromatograph/mass spectrometer was evaluated as an oxidative marker to determine hexanal content in vegetable oils. For the effective analysis, a cooled injection system (CIS) was used to focus and to introduce the hexanal desorbed from the Tenax TA. The temperature of the CIS was maintained at -60 °C for 12 min before desorbing the hexanal. Hexanal was separated on a capillary column (DB-5, 0.25 mm × 60 m, 0.25 µm in film thickness) from 50 to 230 °C, followed by mass spectrometer-selected ion monitoring analysis at m/z 56. The instrumental response to hexanal was highly linear from 10 ng mL(-1) to 1 µg mL(-1) (r(2) = 0.9999). The relative standard deviation (RSD) of intra- and inter-day repeatability was acceptable, with values of less than 3.88 and 4.25%, respectively. The LOD and LOQ of hexanal were determined by gas chromatograph/mass spectrometer-selected ion monitoring to be 3.3 and 9.8 ng mL(-1), respectively. The acid value, peroxide value and fatty acid composition revealed a good correlation with the hexanal concentration.