• Journal of Life Science
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• v.23 no.12
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• pp.1516-1524
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• 2013

To find a novel skin whitening agent, the effect of cordycepin-enriched Cordyceps militaris (CM${\alpha}$) extract fermented by fungi on anti-melanogenesis in B16F0 mouse melanoma cells was investigated. Fermented CM${\alpha}$ was prepared with fungi, including Monascus purpureus (Mp), Aspergillus oryzae (Ao), Aspergillus kawachii (Ak), and Rhizopus oryzae (Ro), respectively. When the content of the phenolics and the flavonoids and the activities of the antioxidant and the mushroom tyrosinase inhibition were measured in the CM fermented by Ak (AkF-CM), the highest content of the phenolics was 46 mg/g dry weight and the highest content of the flavonoids was 0.93 mg/g; the highest activity of the DPPH radical scavenging was 62.74% and the highest activity of the mushroom tyrosinase inhibition was 79.97% CM${\alpha}$CM${\alpha}$. From this result, AkF-CM${\alpha}$ exhibited the highest mushroom tyrosinase inhibitory activity and so it was used in subsequent anti-melanogenesis. B16F0 melanoma cells were treated with 1-10 mg/ml concentrations of AkF-CM${\alpha}$ and 200 ${\mu}M$ arbutin as the positive control. The melanin content and cell viability of the melanoma cells by arbutin treatment decreased to 43% and 92% of the control, respectively. AkF-CM${\alpha}$ treatment at 1, 3, and 5 mg/ml concentrations decreased the extracellular melanin release induced by IBMX treatment by 35%, 45%, and 53%, respectively. AkF-CM${\alpha}$ showed inhibitory activity against both intracellular tyrosinase in melanoma cells and mushroom tyrosinase. AkF-CM${\alpha}$ reduced the protein level of tyrosinase in the IBMX-stimulated cells. These results indicate that AkF-CM${\alpha}$ suppressed the activity and protein content of cellular tyrosinase and decreased the total melanin content in cultured B16F0 melanoma cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.6
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• pp.1499-1505
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• 2007

Peroxynitrite ($ONOO^-$) is a reactive oxidant formed from superoxide anion radical (${\cdot}\;O_2-$) and nitric oxide (NO), which can oxidize cellular components such as essential protein, non-protein thiols, DNA, low-density lipoproteins and membrane phospholipids. ${\cdot}\;O_2-$ and $ONOO^-$ have contributed to the pathogenesis of diseases such as stroke, heart disease, Alzheimer's disease and atherosclerosis. Because of damaging effects of ${\cdot}\;O_2-$ and $ONOO^-$ oxidants, Vespae Nidus, which has been known to strengthen the kidneys to preserve the vital energy. was tested as a potential specific scavenger of those oxidants. In this study, the viability of Vespae Nidus (1, 10, 50 g/ml) to scavenge ${\cdot}\;O_2-$, NO, $ONOO^-$ and so to protect cells against tert-butylhydroxyperoxide (t-BHP) induced cell death was tested. The levels of ${\cdot}\;O_2-$ and $ONOO^-$ were detected by staining with DCFH-DA and DHR 123, respectively. Protein expression levels of COX-2, iNOS and $NF{-\kappa}B$ were assayed by western blot. Vespae Nidus blocked t-BHP-induced cell death in a dose-dependent fashion. Vespae Nidus inhibited t-BHP-induced production of ${\cdot}\;O_2-$, NO and $ONOO^-$ in YPEN cells. The lipid peroxide level was increased and glutathione level was decreased in lipopolysaccharide (LPS)-treated ICR mouse, whereas the ones in the Vespae Nidus-administered group were regulated beneficially. Vespae Nidus inhibited the expression of COX-2, iNOS and NF-κB (p65 and p50) genes in LPS-treated ICR mouse. The present study suggests that Vespae Nidus is a powerful antioxidant and promotes cellular defense activity by scavenging the toxic oxidants such as ${\cdot}\;O_2-$ and $ONOO^-$.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.419-429
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• 1998

Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) has been used for the treatment of allergic disease. IgE is normally one of the least abundant immunoglobulin (Ig) isotypes in the serum of both humans and several species of experimental animals: however a number of different stimuli can result in profound increases in IgE levels relative to other isotypes. In rodents, infection with many parasitic helminths can cause approximately 100-fold elevation in IgE within 2 wks. Immunization of mice with small amounts of protein antigens on alum also results in 10-fold to fold increase in total serum IgE, much of it specific for the immunizing antigen. In this experiment, I investigated the effect of an aqueous extract of Poncirus trifoliata (L.) Raf (Rutaceae) fruits (PTFE) on a in vivo and in vitro IgE production. PTFE dose-dependently inhibited the serum levels of IgE induced by antigens. The regulation of IgE synthesis is influenced by T cells and T cell derived factors. IL -4, a T cell-derived cytokine, has been shown to stimulate murine IgE synthesis both in vitro and in vivo. Current evidence suggests that IL-4 induces IgE synthesis in the mouse by stimulating H chain isotype switch. Lipopolysaccharide (LPS) plus IL-4 cause about l00-fold increase in IgE secretion by murine B cells. The effects of PTFE on the IL-4-dependent IgE response by mouse whole spleen cells were studied. Whole spleen cells were cultured for 7 days in the presence of LPS plus IL-4 and PTFE and the supernatants were assayed for IgE. IL-4 dependent IgE production of LPS-stimulated whole spleen cells was inhibited by PTFE. Moreover, in the present study using U266Bl human IgE-bearing B cells, I found that PTFE inhibited the production of IgE activated by LPS plus IL-4. These results indicate that PTFE have antiallergic activity by inhibition of IgE production from B cells.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.167-171
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• 2010

The present study aims to evaluate Cordyceps militaris water extract (CMWE) with a view to develop melanogenesis inhibitors. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were investigated. CMWE, at $5000\;{\mu}g/ml$, inhibited tyrosinase activity of 71% and DOPA oxidation of 40% as reacting with L-DOPA. Furthermore, B16 mouse melanoma cell survived over 50% from low to high dose on MTT assay, and CMWE markedly inhibited (> 50%) melanin synthesis at $5000\;{\mu}g/ml$. The inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. Key enzyme of melanin biosynthesis is tyrosinase which catalyses a beginning step from tyrosine to DOPA quinine and melanin formation step, respectively. These results indicated that CMWE may be a potential source of novel whitening agents for cosmetic or therapeutic application.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.47-57
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• 2005

Leptin, the product of the obese gene, is a circulating hormone secreted primarily from adipocytes. Several results suggest that leptin is important mediators of bone metabolism. The present study was undertaken to determine the effects of leptin on anti-osteoclastogenesis using murine precursors cultured on Ca-P coated plates and on the production of osteoprotegerin (OPG) in osteoblastic cells. Additionally, this study examined the possible involvement of prostaglandin $E_2\;(PGE_2)$/protein kinase C (PKC)-mediated signals on the effect of leptin on anti-osteoclastogenesis to various culture systems of osteoclast precursors. Osteoclast generation was determined by counting tartrate-resistant acid phosphatase positive [TRAP (+)] multinucleated cells (MNCs). Osteoclastic activity was determined by measuring area of resorption pits formed by osteoclasts on Ca-P coated plate. The number of 1,25-dihydroxycholecalciferol $(1,25[OH]_2D_3)$- or $PGE_2$-induced TRAP (+) MNCs in the mouse bone marrow cell culture decreased significantly after treatment with leptin. The number of receptor activator of NF-kB ligand (RANKL)-induced TRAP (+) MNCs in M-CSF dependent bone marrow macrophage (MDBM) cell or RAW264.7 cell culture decreased significantly with leptin treatment. Indomethacin inhibited osteoclast generation induced by $1,25[OH]_2D_3$ and dexamethasone, however, no significant differences were found in the leptin treated group when compared to the corresponding indomethacin group. Phorbol 12-myristate 13-acetate (PMA), a PKC activator, inhibited osteoclast generation induced by $1,25[OH]_2D_3$. The number of TRAP (+) MNCs decreased significantly with treatment by PMA at concentrations of 0.01 and $0.1{\mu}M$ in culture. Leptin inhibited PMA-mediated osteoclast generation. Isoquinoline-5-sulfonic 2-methyl-1-piperazide dihydrochloride (H7) had no effect on osteoclast generation induced by $1,25[OH]_2D_3$. Cell culture treatment with leptin resulted in no significant differences in osteoclast generation compared to the corresponding H7 group. Indomethacin showed no significant effect on TRAP (+) MNCs formation from the RAW264.7 cell line. PMA inhibited TRAP (+) MNCs formation induced by RANKL in the RAW264.7 cell culture. H7 had no effect on osteoclast generation from the RAW264.7 cell line. There was no difference compared with the corresponding control group after treatment with leptin. $1,25[OH]_2D_3$- or $PGE_2$-induced osteoclastic activity decreased significantly with leptin treatment at a concentration of 100 ng/ml in mouse bone marrow cell culture. Indomethacin, PMA, and H7 significantly inhibited osteoclastic activity induced by $1,25[OH]_2D_3$ in mouse bone marrow cell culture. No significant differences were found between the leptin treated group and the corresponding control group. The secretion of OPG, a substance known to inhibit osteoclast formation, was detected from the osteoblasts. Treatment by leptin resulted in significant increases in OPG secretion by osteoblastic cells. Taken these results, leptin may be an important regulatory cytokines within the bone marrow microenvironment.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1179-1187
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• 2001

In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.6
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• pp.529-539
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• 2008

Background and Purpose : Oral squamous cell carcinoma (OSCC) is one of the most aggressive tumors of the head and neck area. OSCC is known to preferentially metastasize via lymphatic system, and resulting cervical lymph node metastasis is the most reliable of treatment failure. But the biological mechanism of the regional nodal metastasis is not clear. So, we determined metastasis-related factors in orthotopic nude mouse models of OSCC. Experimental Design : Two cell lines-KB and YD-10B cells, established from human oral mucosal squamous cell carcinoma, were xenografted into the tissue space of athymic murine mouth floor. The mice were followed for tumor development and growth, the murine tumors were examined histopathologically for local invasion or regional or distant metastasis. Finally, we performed immunohistochemical assays with antiepithelial growth factor (EGF), EGF receptor (EGFR), phosphorylated EGFR (pEGFR), and anti-vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)-2, phosphorylated VEGFR-2/3 (pVEGFR-2/3) antibodies. We also determined the microvessel density. Results : Transplantation of human OSCC tumor cells into the mouth floor successfully resulted in the formation of orthotopic tumors. KB cell line showed significantly higher tumor proliferation and higher nodal metastatic potential than YD-10B cell line. Furthermore, immunohistochemical staining demonstrated higher expression of EGFR/pEGFR, VEGF, and pVEGFR-2/3 as well as higher microvessel density in KB murine tumors than in YD-10B murine tumors. Conclusion : An orthotopic model of OSCC in athymic mice was established which copies the cervical lymph nodal metastasis of human OSCC. Our mouth floor model should facillitate the understanding of the molecular pathogenesis of cervical nodal metastasis of OSCC.

• BMB Reports
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• v.49 no.10
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• pp.527-528
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• 2016

In embryonic stem cells (ESCs), cell cycle regulation is deeply connected to pluripotency. Especially, core transcription factors (CTFs) which are essential to maintaining the pluripotency transcription programs should be reset during M/G1 transition. However, it remains unknown about how CTFs are governed during cell cycle progression. Here, we describe that the regulation of Oct4 by Aurora kinase b (Aurkb)/protein phosphatase 1 (PP1) axis during the cell cycle is important for resetting Oct4 to pluripotency and cell cycle related target genes in determining the identity of ESCs. Aurkb starts to phosphorylate Oct4(S229) at the onset of G2/M phase, inducing the dissociation of Oct4 from chromatin, whereas PP1 binds Oct4 and dephosphorylates Oct4(S229) during M/G1 transition, which resets Oct4-driven transcription for pluripotency and the cell cycle. Furthermore, Aurkb phosphormimetic and PP1 binding-deficient mutations in Oct4 disrupt the pluripotent cell cycle, lead to the loss of pluripotency in ESCs, and decrease the efficiency of somatic cell reprogramming. Based on our findings, we suggest that the cell cycle is directly linked to pluripotency programs in ESCs.

• The Korea Journal of Herbology
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• v.30 no.1
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• pp.95-101
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• 2015

Objectives : The aim of this study was to confirm whether or not coral has a preventive effect on development of atopic dermatitis induced by house mite(dermatophagoides pteronyssinus) in NC/Nga mice. Methods : This study was undertaken by using a reliable Atopic dermatitis mouse model demonstrating similar immune response. Lobophytum crassum was administered orally to NC/Nga mouse for 3 weeks. In order to verify the effectiveness of Lobophytum crassum in atopic dermatitis treatment, its role in immune factors were observed in NC/Nga mice. Results : ALT, AST, BUN and creatine levels were all within in the normal ranges in MC-1 200 and 400 (mg/kg) treated groups, indicating no induced toxicity. MC-1 200 and 400 (mg/kg) groups decreased of atopic dermatitis skin manifestation in NC/Nga mouse of MC-1 200 and 400 (mg/kg) groups compared to that of the control group and decreased the ratio of WBC and lymphocyte in blood. Also, MC-1 200 and 400 (mg/kg) groups significant decreased the ratio of CD4+, CD8+, CD11b+/Gr1+, B220/CD23 and CD4/CD25 immune cell ratio in ALN. Finally MC-1 200 and 400 (mg/kg) groups significantly increased the ratio of CD4+, CD8+, B220/CD23 and CD4/CD25 immune cells in DLN. Conclusions : Theses results suggested that Lobophytum crassum has suppressive effects on aberrant and overactive immunological activities in dermatophagoides pteronyssinus-induced dermatitis mice of NC/Nga.

• The Journal of Korean Medicine
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• v.31 no.2
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• pp.19-35
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• 2010

Objectives: This study was carried out to find the effects of Changchuldoin-tanggamibang (hereinafter referred to CDIT) on the inhibition of arthritis induced by collagen on DBA/1J mouse. Methods: The experimental mice were divided into four groups: normal group (Nr), control group (CIA-CT), methotrexate group (CIA-MTX), and Changchuldoin-tanggamibang group (CIA-CDIT). Cytotoxicity, hepatotoxicity, arthritis index, value of immunocytes in draining lymph node and paw joint, and rheumatoid factor (IgG, IgM) in serum were measured in vivo. Results: 1. Cytotoxicity against hFCs was not shown in any concentration. 2. Hepatotoxicity was low in the CDIT-treated group compared with the MTX group. 3. The arthritis index decreased significantly. 4. In total cell counts of DLN and paw joint, the cells in DLN increased significantly while there was a significant decrease in paw joint. 5. In lymph nodes, CD19+, CD3+, CD4+, CD8+, CD3+/CD8+, CD3+/CD69+, CD4+/CD25+, CD3+/CD49b+, and CD4+/CD44+ cells increased significantly, while B220+/CD23+, and CD11c+/MHCII+ cells decreased significantly. 6. In joints, CD3+, CD4+, CD4+/CD25+, and CD11b+/Gr-1+ cells decreased significantly. 7. The level of IgG decreased and the level of IgM significantly decreased compared with the control. 8. Anti-collagen II in serum decreased compared with the control. 9. Around the joint of the CDIT group, infiltration of inflammation, synovial hyperplasia, invasion of cytokine, of cartilage, deposition of collagen and synovial injury decreased compared with the control in histopathologic observation (HE, MT staining). Conclusions: Comparison of the results for this study showed that CDIT had immunomodulatory effects. We expect that CDIT could be used as a effective drug for not only rheumatoid arthritis but also another auto-immune diseases. Therefore, we have to survey continuously, looking for effective substances and mechanisms in the future.