• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2002.11a
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• pp.14-40
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• 2002

15-Deoxy-${\Delta}^{12, 14}$-prostaglandin $J_2$ (15-deoxy-$PGJ_2$), a naturally occurring ligand activates the peroxisome proliferator-activated $receptor-{\gamma}(PPAR-{\gamma}$). Activation of $PPAR-{\gamma}$ has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-$PGJ_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-$PGJ_2$ (0.2 to 1.6 ${\mu}M$) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/ml). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-$PGJ_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-$PGJ_2$(0.8 ${\mu}M$) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-$PGJ_2$ on the expression of p38 MAP kinase and activation of AP-1, The promoting ability of 15-deoxy-$PGJ_2$ did not occur through $PPAR-{\gamma}$, as synthetic PPAR-${\gamma}$ agonist andantagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), $PPAR-{\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and $PGE_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of f 5-deoxy-$PGJ_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-$PGJ_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 single pathway may be important in the promoting activity of 15-deoxy-$PGJ_2$ cells.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.11b
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• pp.14-40
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• 2002

15-Deoxy- Δ$\^$12,14/-prostaglandin J$_2$ (15-deoxy-PGJ$_2$), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$). Activation of PPAR-y has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-PGJ$_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-PGJ$_2$ (0.2 to 1.6 ${\mu}$M) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/$m\ell$). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-PGJ$_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-PGJ$_2$ (0.8 ${\mu}$M) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-PGJ$_2$ on the expression of p38 MAP kinase and activation of AP-1. The promoting ability of 15-deoxy-PGJ$_2$ did not occur through PPAR-${\gamma}$, as synthetic PPAR-${\gamma}$ agonist and antagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), PPAR-${\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and PGE$_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of 15-deoxy-PGJ$_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-PGJ$_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 signal pathway may be important in the promoting activity of 15-deoxy-PGJ$_2$ on the differentiation of PC12 cells.

• The Korea Journal of Herbology
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• v.31 no.3
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• pp.59-69
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• 2016

Objectives： Presently Mume Fructus (MF) undergoes fumigation, which produces benzo[a]pyrene. As a primary analysis with the aims to minimize the production of benzo[a]pyrene and to suggest standards for processing the MF, the steaming method was chosen among the various processing methods, and reviewed through a series of experiments.Methods： Methods：Pitted and un-pitted MF were steamed and processed into samples. After testing level of benzo[a]pyrene, the samples were analyzed for amount of polyphenol and flavonoids. Scavenging activities of the samples for the DPPH and ABTS radicals were tested. In order to measure anti-inflammatory effects of the samples, cell survival rate was investigated using CCK-8 Assay. Also, water extracts of dried and steamed MF were administered to the RAW 264.7 cells to compare expressions of NO, PGE₂, IL-1β, and TNF-α. In addition, anti-diarrhea effects of the herbal medicine were tested on animal models with diarrhea induced by MgSO₄ and Castor oil.Results： Regardless of pitting, processed MF contained no benzo[a]pyrene. Anti-oxidation effect increased in relation to the frequency of steaming process. However, extracts of dried and steamed MF suppressed different kinds of inflammation factors, and extract of dried MF showed superior anti-diarrhea effect than extract of steamed MF.Conclusions： It is suggested that steaming method of MF is recommended for processing the herbal medicine without the production of benzo[a]pyrene. But regarding that dried and steamed MF showed differences in their anti-oxidative, anti-inflammatory, and anti-diarrhea effects, it is recommended to perform further researches on different efficacies of MF according to their processing methods.

• IMMUNE NETWORK
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• v.10 no.5
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• pp.145-152
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• 2010

Background: The flowers of Inula japonica (Inulae Flos) have long been used in traditional medicine for the treatment of inflammatory diseases. In the present study, we investigated the anti-inflammatory properties of Inulae Flos Extract (IFE). Methods: The anti-inflammatory effects of IFE against nitric oxide (NO), $PGE_2$, TNF-${\alpha}$, and IL-6 release, as well as NF-${\kappa}B$ and MAP kinase activation were evaluated in RAW 264.7 cells. Results: IFE inhibited the production of NO and the expression of inducible nitric oxide synthase (iNOS) in LPS-stimulated RAW264.7 cells. In addition, IFE reduced the release of pro-inflammatory cytokines, such as TNF-${\alpha}$ and IL-6. Furthermore, IFE inhibited the NF-${\kappa}B$ activation induced by LPS, which was associated with the abrogation of $I{\kappa}B-{\alpha}$ degradation and subsequent decreases in nuclear p65 and p50 levels. Moreover, the phosphorylation of ERK, JNK, and p38 MAP kinases in LPS-stimulated RAW 264.7 cells was suppressed by IFE in a dose-dependent manner. Conclusion: These results suggest that the anti-inflammation activities of IFE might be attributed to the inhibition of NO, iNOS and cytokine expression through the down-regulation of NF-${\kappa}B$ activation via suppression of $I{\kappa}B{\alpha}$ and MAP kinase phosphorylation in macrophages.

• Biomedical Science Letters
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• v.26 no.3
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• pp.157-163
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• 2020

Long-term exposure to environmental pollutants can impair the human skin's barrier function and promote skin aging mechanisms. The visible consequences of these effects are dryness, wrinkles, black spots and worsening skin sensitivity. As awareness of the effects of environmental stressors on the skin has recently increased, consumers' demand for cosmetics that can provide anti-pollution effects is increasing. In this study, the possibility of anti-pollution cosmetic material was investigated by measuring the inhibitory effect of free radicals using a mixture of Nypa fruticans Wurmb (NF), Saussurea neoserrata (SN), Codium fragile (CF) and Enteromorpha compressa (EC), which has anti-inflammatory and antioxidant effects against irritation caused by dust. Combining the experimental results of a mixture of NF, SN, CF and EC by induction of fine dust stimulation, inhibition of inflammatory factors (PGE₂) and intracellular free radicals (ROS) by inhibiting effects were significant, indicating the possibility of use as anti-pollution cosmetics. The most plants used as sources of anti-pollution cosmetic ingredients contain antioxidants as active substances. In our highly industrialized and chemically polluted world, it is not surprising that most plants used as sources of anti-pollution cosmetic ingredients contain antioxidants as active substances. Considering our results, it can be suggested that this anti-pollutant consisting of NF, SN, CF and EC may be a good ingredient for skincare products for the cosmetic industry due to their antioxidant properties, which may especially alter skin aging.

• Korean Journal of Pharmacognosy
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• v.43 no.3
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• pp.217-223
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• 2012

In the present study, we investigated anti-inflammatory activity of artemisinic acid in HaCaT cells and RAW264.7 cells. Artemisinic acid showed inhibitory activity on macrophage-derived chemokines (MDC) expression, a factor related with atopic dermatitis (AD), in interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$-stimulated HaCaT cells. In the study on action mechanism, pretreated artemisinic acid reduced the phosphorylation of STAT1 and p38 and the degradation of $I{\kappa}B$ by IFN-${\gamma}$ and TNF-${\alpha}$ stimulations. However, artemisinic acid didn't show the inhibitory activity on LPS-induced inflammatory mediators (NO, $PGE_2$, IL-6) in RAW264.7 cell. These results indicate that artemisinic acid inhibits IFN-${\gamma}$ and TNF-${\alpha}$-induced MDC expression through inhibition of signal factors, STAT1, NF-${\kappa}B$, and p38, in HaCaT keratinocytes.

• Journal of People, Plants, and Environment
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• v.21 no.6
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• pp.485-492
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• 2018

This study aimed to develop and validate highly sensitive determination method of a prostaglandin ($PGE_1$, OP 1206) in human plasma by LC-MS/MS using column switching. Plasma stored at $-30^{\circ}C$ and treated with methanol effectively inhibited interferences synthesized post-sampling. Samples were added with internal standard and were separated by reversed-phase HPLC with a cycle time of 30min. The method was selective for OP 1206 and the regression models, based on internal standard, were linear across the concentration range 0.5-50 pg/mL with the limit of quantification of 0.5 pg/mL (limit of quantitation, LOQ) for OP 1206. The calibration curve of OP 1206 standards spiked in five individual plasma samples was linear ($r^2=0.9999$). Accuracy and precision at the concentrations of 0.5, 1.5, 5.0 and 40 pg/mL, and at the lower LOQ of 0.5 pg/mL were excellent at 20%. OP120 < 6 was stable in plasma samples for at least 24 hours at room temperature, 24 hours frozen at $-70^{\circ}C$, 24 hours in an auto sampler at $6^{\circ}C$, and for two freeze/unfreezing cycles. The validated determination method successfully quantified the concentrations of OP 1206 in plasma samples from simulated administrating a single $5{\mu}g$ OP 1206 formulation. Thus, this novel LC-MS/MS technique for drug separation, detection and quantitation is expected to become the standard highly-sensitive detection method in bioanalysis and to be applied to many low dose pharmaceutical products.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.35 no.3
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• pp.36-47
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• 2022

Objectives : The purpose of this study was to evaluate the anti-Inflammatory effect of Hwadokdan(HDD) extracted with water. Methods : We pretreated LPS-activated Raw 264.7 cell with HDD and anti-inflammatory effect of HDD is investigated by measuring NO production, relative iNOS, and TNF-α, IL-6, IL-1β, PGE2 productions level, p-IκBα, IκBα, NFκB. Results : HDD reduced NO production and iNOS protein, TNF-α, IL-6 level at concentration of 100㎍/㎖ and 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. There wasn't a significant cytotoxicity in MTT assay. And HDD reduced IL-1β level at concentration of 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. HDD reduced p-IκBα and NFκB protein level at concentration of 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. And HDD enhanced IκBα protein level at concentration of 100㎍/㎖ and 300㎍/㎖ in LPS-activated Raw 264.7 cell significantly. Conclusions : These suggests that HDD can be used as a therapeutic drug for various inflammatory diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.1
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• pp.18-25
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• 2017

This study describes the preliminary evaluation of antioxidant and anti-inflammatory activities of Allium hookeri. A. hookeri was extracted using crude extract and then fractionated sequentially with n-hexane, $CH_2Cl_2$, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, we first examined the inhibitory effect of A. hookeri extracts on production of oxidant stresses (2,2-diphenyl-1-picrylhydrazyl, xanthine oxidase, and superoxide). In addition, we examined the inhibitory effects of A. hookeri on production of pro-inflammatory factors (nitric oxide, prostaglandin $E_2$, inducible nitric oxide synthase, and cyclooxygenase-2) in murine macrophage RAW 264.7 cells stimulated with lipopolysaccharide. Of the sequential solvent fractions of A. hookeri, EtOAc fractions showed decreased production of oxidant stresses, and $CH_2Cl_2$ and EtOAc fractions of A. hookeri inhibited production of pro-inflammatory factors. EtOAc fraction inhibited production of pro-inflammatory cytokines (interleukin-6 and -$1{\beta}$). These results suggest that A. hookeri has significant effects on oxidant stresses and pro-inflammatory factors and is a possible antioxidant and anti-inflammatory therapeutic and preventive material.

• Molecules and Cells
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• v.40 no.3
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• pp.222-229
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• 2017

Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.