• Journal of Radiation Industry
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• v.5 no.1
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• pp.15-20
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• 2011

Protein kinases are the most important drug targets for the treatment of numerous diseases. The involvement of protein kinase C (PKC) in many biological processes such as development, memory, cell differentiation, and proliferation has been demonstrated. PKC is recognized as an important player in carcinogenesis. Thus, a variety of PKC inhibitors have been investigated. Among them, flavonoids have been demonstrated to affect the activity of many mammalian in vitro enzyme systems. The recent investigation was performed to evaluate the inhibitory effects of hesperidin, which is a flavonoid, on the proliferation and carcinogenesis of many cancers. In this study, an efficient kinase assay based on a biochip using radio-phosphorylation was established and performed for an examination of the inhibitory effects of hesperidin on PKC activity at different concentrations of 50, 200, 500 nM. It was found that hesperidin shows inhibitory potency on PKC, and that the biochip can be used to rapidly screen kinase inhibitors resulting in the therapeutic agents.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Journal of Dental Anesthesia and Pain Medicine
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• v.22 no.5
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• pp.369-376
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• 2022

Background: Nonobstetric surgery is sometimes required during pregnancy, and neck abscess or facial bone fracture surgery cannot be postponed in pregnant women. However, dental surgery can be stressful and can cause inflammation, and the inflammatory response is a well-known major cause of preterm labor. Propofol is an intravenous anesthetic commonly used for general anesthesia and sedation. Studies investigating the effect of propofol on human amnion are rare. The current study investigated the effects of propofol on lipopolysaccharide (LPS)-induced inflammatory responses in human amnion-derived WISH cells. Methods: WISH cells were exposed to LPS for 24 h and co-treated with various concentrations of propofol (0.01-1 ㎍/ml). Cell viability was measured using the MTT assay. Nitric oxide (NO) production was analyzed using a microassay based on the Griess reaction. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE 2), p38, and phospho-p38 was analyzed using western blotting. Results: Propofol did not affect the viability and NO production of WISH cells. Co-treatment with LPS and propofol reduced COX-2 and PGE₂ protein expression and inhibited p38 phosphorylation in WISH cells. Conclusion: Propofol does not affect the viability of WISH cells and inhibits LPS-induced expression of inflammatory factors. The inhibitory effect of propofol on inflammatory factor expression is likely mediated by the inhibition of p38 activation.

• The Korea Journal of Herbology
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• v.28 no.5
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• pp.113-119
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae Gigantis Radix Water Extract(AG) on the production of proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide(LPS). Method : RAW 264.7 cells were cotreated with AG(50 and 100 ug/mL) and lipopolysaccharide(LPS; 1 ug/mL) for 24 hours. After 24 hour treatment, using Bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, IL-$1{\beta}$, IL-10, tumor necrosis factor-alpha(TNF-${\alpha}$), granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), interferon inducible protein-10(IP-10), leukemia inhibitory factor(LIF), lipopolysaccharide-induced chemokine(LIX), monocyte chemoattractant protein-1(MCP-1), macrophage colony-stimulating factor(M-CSF), macrophage inflammatory protein(MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-2, Regulated on Activation, Normal T cell Expressed and Secreted(RANTES) and vascular endothelial growth factor(VEGF) were measured. Result : AG significantly inhibited LPS-induced production of TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, and M-CSF from LPS-stimulated RAW 264.7 cells at the concentrations of 50 and 100 ug/mL. AG significantly inhibited LPS-induced production of MIP-$1{\beta}$, MIP-2, GM-CSF, and IL-6 from LPS-stimulated RAW 264.7 cells at the concentrations of 50 ug/mL. AG significantly inhibited LPS-induced production of VEGF from LPS-stimulated RAW 264.7 cells at the concentrations of 100 ug/mL. But AG did not show any significant effect on the production of MCP-1, LIF, LIX, IP-10 and IL-$1{\beta}$ from LPS-induced RAW 264.7 cells. Conclusion : These results suggest that AG has anti-inflammatory effect related with its inhibition of proinflammatory mediators such as TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, MIP-$1{\beta}$, MIP-2, GM-CSF, IL-6, VEGF and M-CSF in LPS-induced macrophages.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.40-40
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• 2023

Atopic dermatitis is a chronic inflammatory skin diseases caused by skin barrier dysfunction. Allium victoralis var. Platyphyllum (AVP) is a perennial plant used as vegetable and herbal medicine. The purpose of this study was to suggest that AVP is a new cosmetic material by examining the effects of AVP on the skin barrier and inflammatory response. A bibliometric network analysis was performed through keyword co-occurrence analysis by extracting author keyword from 69 articles retrieved from SCOPUS. We noted the anti-inflammatory activity shown by the results of clustering and mapping from network visualization analysis using VOSviewer software tool. HPLC-UV analysis showed that AVP contains 0.12 ± 0.02 mg/g of chlorogenic acid and 0.10 ± 0.01 mg/g of gallic acid. AVP at 100 ㎍/mL was shown to increase the mRNA levels of filaggrin and involucrin related to skin barrier function by 1.50-fold and 1.43-fold, respectively. In the scratch assay, AVP at concentrations of 100 ㎍/mL and 200 ㎍/mL significantly increased the cell migration rate and narrowed the scratch area. In addition, AVP suppressed the increase of inflammation-related factors COX-2 and NO and decreased the release of β-hexosaminidase. This study suggests that AVP can be developed as a functional cosmetic material for atopy management through skin barrier protection effects, anti-inflammatory and anti-itch effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.63-71
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• 2004

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98％ at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97％ at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79％ at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30％. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.7 no.1
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• pp.77-97
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• 2001

Anti-angiogenesis is one of therapies which have been high-lightened on the research of cancer treatment. Anti-angiogenesis means that new blood vessels are created from a existing capillary tube and it is a important process on metastasis and permeation when cancer is created or formed. Since angiogenesis have been under research, a complete recovery oriented treatment against cancer have been suggested blocking metastasis, delaying the growth of cancer cell, and blocking the supply of oxygen and nutritive substance through the web of blood vessels. Until now, there are several anti-angiogenesis, which have been known to the public, such as thalidomide, angiostatin, endostatin, 2-methoxyestradiol, TNP-470, and marimastat, etc. Additionally, 17 clinical testing projects about anti-angiogenesis are on the process in NCI(National Cancer Institute). Especially, TNP-470 showed effectiveness against cancer on clinical testing after finishing animal testing. Based on existing researches showing that Yinsamyangwui-tang is effective to strengthening body resistance and Whallakhyolenyng-dan effects cells on the inside of blood vessel because Whallakhyolenyng- dan restrains cell adhesion during the restraining period of a blood vessel, I tried to research the effect of Whalakhyolenyng-dan plus Yinsamyangwui-tang on angiogenesis. I made a conclusion putting into operation through using SK-Hep-1 (KCLB 30052), A549(KCLB 10185), AGS(KCLB 21739), and BCE(Bovine Capillary Endothelial Cell). Followings are the results of my experimental research: 1. According to the researching results of anti-cancer activation against cancer cell, Whallkhyoleyng dan plus Yinsamyangwui-tang decreased the number of cancer cells -- While injecting $600{\mu}g/ml$, injected groups decreased 3.1% more comparing with the contrastive group of SK-Hep-1, 49.7% more comparing with the contrastive group of A549, and 31.0% more comparing with the contrastive group of AGS. 2. According to the researching results of DNA composition effect between BCE and cancer cell, Whallakhyoleyng-dan plus Yinsamyangwui-tang reduced the rate of SK-Hep-1 synthesis inhibition by 59.1% at $600{\mu}g/ml$ intensity comparing with contrastive group; for A549, 72.6%; for AGS, 6.1%, for BCE, 28.9%. 3. According to the researching results about the effect of BCE cell to angiogenesis, angiogenesis was restrained at $400{\mu}g/ml$ intensity during 18 hours observation. 4. In the case of aortic ring assay, the half level of angiogenesis was reduced comparing with the contrastive group while injecting with $400{\mu}g/ml$ intensity; with $800{\mu}g/ml$, under 10% comparing with contrastive group; and with $1600{\mu}g/ml$, complete restrain. According to the above results, Whallakhyoleyng-dan plus Yinsamyangwui-tang was proved to have an anti-angiogenetic effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.1
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• pp.65-72
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• 2009

Skin aging appears to be principally attributed to a decrease in both levels of Type I collagen and regeneration ability of dermal fibroblasts. It is important to introduce an efficient and safe agent for effective management of skin aging. To this end, we performed screening for anti-ageing agents and then found that vegetable peptones (pea and wheat) promoted cell proliferation of adult stem cells. Vegetable peptones may be considered as useful medium additives because it can supply nutrients, peptides, amino acids or growth factor analogues. This study was designed to investigate effects of vegetable peptones on cell proliferation/collagen production and their possible mechanisms in human dermal fibroblasts. In cell proliferation assay, vegetable peptones significantly promoted cell proliferation in a concentration-dependent manner. In addition, human COL1A2 promoter luciferase and type I procollagen synthesis assays showed that vegetable peptones induce type I procollagen production through the activation of COLlA2 promoter. In both TGF-${\beta}1$ luciferase reporter and ELISA assays, vegetable peptones was found to induce TGF-${\beta}1$ production, suggesting that vegetable peptones induce type I procollagen production through the activation of TGF-${\beta}1$. When applied topically in a human skin twice a day for an 4-week period of time, vegetable peptones did not induce any adverse reactions. Theretore, based on these results, we suggest the possibility that vegetable peptones may be considered as an attractive, wrinkle-reducing candidate for topical application.

• Journal of Life Science
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• v.26 no.6
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• pp.633-639
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• 2016

Biomaterials including polymer, metal, ceramic, and composite have been widely applied for medical uses as medical fibers, artificial blood vessels, artificial joints, implants, soft tissue, and plastic surgery materials owing to their physicochemical properties. However, the biocompatibility and toxicity for film- and plate-form biomaterials is difficult to measure in mammalian cells because there is no appropriate incubation system. To solve these problems, we developed a novel mammalian cell culture system consisting of a silicone ring, top panel, and bottom panel and we applied two polymer films (PF) and one metal plate (MP). This system was based on the principal of sandwiching a test sample between the top panel and the bottom panel. Following the assembly of the culture system, SK-MEL-2 cells were seeded onto Styela Clava Tunic (SCT)-PF, NaHCO₃-added SCT (SCTN)-PF, and magnesium MP (MMP) and incubated at 37℃ for 24 hr and 48 hr. An MTT assay revealed that cell viability was maintained at a normal level in the SCT-PF culture group at 24 or 48 hr, although it rapidly decreased in the SCTN-PF culture group at 48 hr. Furthermore, the cell viability in the MMP culture group was very similar to that of the control group after incubation for 24 hr and 48 hr. Together, these results suggest the sandwich-type mammalian culture system developed here has the potential for the evaluation of the biocompatibility and toxicity of cells against PF- and MP-form biomaterials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.8
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• pp.1128-1136
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• 2015

This study investigated the anti-inflammatory effects of ethanol extract from Grateloupia elliptica Holmes (GEHEE) on the lipopolysaccharide-induced inflammatory response. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There were no cytotoxic effects on proliferation of macrophages treated with GEHEE compared to the control. GEHEE remarkably suppressed NO and pro-inflammatory cytokines (interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$) production and reduced expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) proteins in a dose-dependent manner. GEHEE also significantly reduced activation of mitogen-activated protein kinases (MAPKs). The formation of edema in mouse ears was reduced at the highest dose compared to the control. GEHEE also reduced dermal thickness and mast cell numbers based on histological analysis. These results suggest that GEHEE exerts significant anti-inflammatory activity via inhibition of $NF-{\kappa}B$ and MAPKs activation and may be a potential anti-inflammatory therapeutic material.