• IMMUNE NETWORK
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• v.6 no.1
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• pp.33-42
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• 2006

Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

• The Korean Journal of Mycology
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• v.35 no.2
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• pp.101-108
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• 2007

Armillaria tabescens, one of edible and medicinal mushrooms belonging to Agaricales of Basidiomycota, has been known to have outstanding curative effects on chronic hepatitis and cholecystitis and inhibitory effects on the sarcoma 180 and Erhrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr, NaCl, Fr. HW and Fr, MeOH, respectively) were extracted from fruiting body of the mushroom. In vitro cytotoxicity tests showed that crude polysaccharides were not cytotoxic against cancer cell lines such as NIH3T3 and Sarcoma 180 at the concentration of $2000\;{\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of $28.8{\sim}46.5%$ in mice inoculated with Sarcoma 180, respectively. Fr. NaCl improved the immunopotentiation activity of B lymphocyte by increasing the alkaline phosphatase activity by $1.8{\sim}2.1$ folds, respectively. In case of Fr, NaCl, the numbers of peritoneal exudate cells and circulating leukocytes were increased by 9 and 1.9 folds, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.3
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• pp.287-291
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• 2009

In order to use Corni fructus as functional food materials, we investigated the biological activities of ethanol extracts from Corni fructus (EECF). The hydrogen-donating activity of EECF was increased in a dose dependent manner compared with untreated control, and the activities by EECF were 64 and 74% at 300 and $500{\mu}g/mL$ concentration, respectively. The NO productions in the RAW264.7 marcrophage cells treated with EECF were increased in dose dependent manners. EECF significantly inhibited the growth of MCF-7 human breast cancer cells in dose and time dependent manners. EECF of $500{\mu}g/mL$ concentration inhibited the proliferation by over 60% in the MCF-7 cells when treated for 72 hr. Also, the proliferations were increased in the MCF-7 cells cultured in the charcoal-treated FBS (cFBS) medium with environmental hormones such as bisphenol or $17{\beta}$-estradiol of $0.1{\mu}M$ whereas the proliferations were decreased in the MCF-7 cells treated with the environmental hormones after treatment of EEFC for 72 hr. The results suggest that Corni fructus would be used as functional food materials.

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.71-81
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• 2005

Objective : Dear antler (Cervus korean TEMMINCK var. mantchuricus Swinhoe) used for traditional immunosuppressive and immuno-activating action. The effect of deer antler herbal-acupuncture(DAH) solution, prepared by water extract method, on cathepsin activities in bone tissues (cartilage and synovial) cells from mouse rheumatoid arthritis (RA) model was studied. The cysteine endoprotease cathepsin mediates degradation of the MHC class II invariant chain (Ii) in human and mouse antigen-presenting cells. The studies described here examine the functional significance of cathepsin inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for RA. Methods : An animal model for RA in BALB/c mice thymectomized 3 days after birth (3d-Tx) was constructed All 3d-Tx BALB/c mice developed autoimmune lesions in the bone tissue cells, starting at 3 weeks of age, and the disease mediated by CD4+ T cells was chronic and progressive. Significant inhibitory effects of DAH solution on cathepsin S and L were observed in each organ in a dose-dependent manner. Moreover, we confirmed that cathepsin S and L activity in each organ were clearly inhibited by DAB solution. When we examined the inhibitory effects of DAH solution against autoantigen-specific T cell responses in vitro, in regional lymph node cells, but not in spleens, from model mice, a significant inhibitory effect of DAB solution was observed in a dose-dependent manner. DAH solution do not block T cell proliferation to Con A, indicated that the dose of DAB solution 10 to $20\;{\mu}g/m{\ell}$ was sufficient to inactivate the autoantigen-specific T cell responses in vitro. In vivo therapeutic effects of DAB solution were examined in a murine model for RA, autoantigen-specific (C-II-specific) T cell response were significantly inhibited in LNCs from DAH solution-treated mice. Results : Iinhibition of cathepsin S and L in vivo alters autoantigen presentation and development of organ-specific autoimmunity in RA model. Conclusion : These data identify selective inhibition of cysteine protease cathepsin S and L as a potential therapeutic strategy for autoimmune disease process such RA. Thus, DAH solution will served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

• The Korean Journal of Mycology
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• v.34 no.2
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• pp.98-104
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• 2006

Armillaria mellea, one of edible and medicinal mushroom belonging to Tricholomataceae of Basid-iomycota, has been known to have outstanding inhibitive effects on the sarcoma 180 and Erhrlich carcinoma of mice. Neutral salt soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr, HW and Fr. MeOH, respectively) were extracted from the mushroom. In vitro cytotoxicity tests, crude polysaccharide were not cytotoxic against cancer cell lines such as NIH3T3 and Sarcoma 180 at the concentration of $1000{\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of $60{\sim}67.5%$ in mice inoculated with Sarcoma 180, respectively. Fr, NaCl improved the immunopotentiation activity of B lymphocyte by increasing the alkaline phosphatase activity by $1.8{\sim}3.0$ folds, respectively. In case of Fr. NaCl, the numbers of peritoneal exudate cells and circulating leukocytes were increased by 10 and 2 folds, respectively.

• Journal of Applied Biological Chemistry
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• v.59 no.2
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• pp.113-124
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• 2016

Red ginseng is known to have many beneficial effects. Cisplatin, an effective antineoplastic drug, can cause many side effects like irreversible sensorineural hearing loss and serious tinnitus in humans. This study is aimed to reduce a cisplatin's side effect, nephrotoxicity by fermentated korean red ginseng. Korea ginseng was produced by steaming and dring and fermentation. And mice were divided into 4 groups- (A) normal mice, (B) Vehicle treated cisplatin mice, (C) RG0F0-treated cisplatin mice, (D) RG8F3-treated cisplatin mice. C and D groups were feed each material 200 mg/kg/day during 4 days. And cisplatin 20 mg/kg injected to B, C, and D groups as abdominal injection. After 24 h, blood sample was collected. The kidneys were harvested for histological, immuno histochemical and western blot analysis. 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity was depended on steaming hours. RG0F0 and RG8F3 (ginseng-8 h steamed and fermented by Saccharomyces cerevisiae) were showed antioxidants effect in DPPH and ABTS radical scavenging activity. Component amounts according to steaming hours. 8 h steamed red ginseng had the most ingredients of ginsenoside. Treatments with RG8F3 reduced cisplatin-induced nephrotoxicity in the mice resulting in increase of GSH and decrease of ROS, BUN, creatinine, and inflammatory mediators. This result seems to be involved with the restriction of the inflammation in the kidney. Therefore, fermented red ginseng might have therapeutic efficacy in reduce kidney injury induced by cisplatin treatment.

• Korean Journal of Medicinal Crop Science
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• v.17 no.3
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• pp.210-216
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• 2009

This study was performed to compare the effects of immuno-modulating activities of Rhodiola sachalinensis A. Bor. fractionized by consecutive solvent separation. The Cytotoxicity of all fractionized extracts on human kidney cell (HEK293) was lower than crude extracts. Generally, the butanol and chloroform extracts showed less cytotoxicity on about 10.07% and 9.67% than the crude extracts. For human immune B and T cell growth, chloroform fraction showed the highest cell growth compared to the control. The secretion of cytokines (IL-6, $TNF-\alpha$) on human B and T cells were increased by adding chloroform extracts. Also, NK cell growth was significantly improved up to nearly 30% by adding the supernatant of B cell medium grown with the chloroform fraction. It was also found that chloroform fraction could yield higher nitric oxide production from macrophage than untreated control cells. Differentiation of HL-60 cells was increased up to 131.9% after treatment with chloroform fraction extracts, compared to the control. These results indicate that the chloroform fraction of R. sachalinensis have high immune activation activity than others fractions and the crude extracts, implying that this chloroform fractions could be used a new functional material.

• Journal of Life Science
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• v.25 no.6
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• pp.703-708
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• 2015

Hox genes encode a highly conserved family of homeodomain-containing transcription factors controlling vertebrate pattern formation along the anteroposterior body axis during embryogenesis. Retinoic acid (RA) is a key morphogen in embryogenesis and a critical regulator of both adult and embryonic cellular activity. Specifically, RA regulates Hox gene expression in mouse- or human-derived embryonic carcinoma (EC) cells. Histone modification has been reported to play a pivotal role in the process of RA-induced gene expression and cell differentiation. As histone modification is thought to play an essential role in RA-induced Hox gene expression, we examined RA-induced initiation of collinear expression of Hox genes and the corresponding histone modifications in F9 murine embryonic teratocarcinoma (EC) cells. Hox expression patterns and histone modifications were analyzed by semiquantitative RT-PCR, RNA-sequencing, and chromatin immuno-precipitation (ChIP)-PCR analyses. The Hoxc4 gene (D0) was initiated earlier than the Hoxc5 to –c10 genes (D3) upon RA treatment (day 0 [D0], day 1 [D1], and day 3 [D3]). The Hox nonexpressing D0 sample had a strong repressive marker, H3K27me3, than the D1 and D3 samples. In the D1 and D3 samples, reduced enrichment of the H3K27me3 marker was observed in the whole cluster. The active H3K4me3 marker was closely associated with the collinear expression of Hoxc genes. Thus, the Hoxc4 gene (D1) and all Hoxc genes (D3) expressed H3K4me3 upon transcription activation. In conclusion, these data indicated that removing H3K27me3 and acquiring H3K4me3 regulated RA-induced Hoxc gene collinearity in F9 cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.4
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• pp.287-292
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• 2009

Human skin is constantly exposed to environmental irritants such as smoke, chemicals and ultraviolet (UV). Free radicals and reactive oxygen species (ROS) caused by these environmental irritants play critical roles in cellular damage. In this study, to investigate the skin cell protective effect of Ophioglossum vulgatum extract, we investigated its effects on intercellular antioxidative activity and UVA-induced MMP expression in human dermal fibroblasts (HDFs). The dried O. vulgatum was extracted in a mixture of ethanol and water (1 : 1) for 24 h at room temperature. The extract was filtered and concentrated in vacuo and lyophilized. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB $20\;mJ/cm^2$. After treatment of O. vulgatum extracts, intracellular ROS levels were measured by luminescence spectrophotometer. Enzyme linked immuno sorbent assay (ELISA), and RT-PCR techniques were used for evaluating the effects of O. vulgatumon on MMP protein and mRNA expression in UVA irradiated HDFs. O. vulgatum extract was found to have ROS scavenging activity with the $IC_{50}$ values of $18.2\;{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system. After treatment of O. vulgatum extracts, the oxidation of CM-DCFDA was inhibited effectively and O. vulgatum extracts showed a potent free radical scavenging activity by 30.4 % at $100\;{\mu}g/mL$ in UVB-irradiated HDFs. UVA induced MMP protein expression was reduced 37.7 % by treatment with O. vulgatum extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Taken together, these results suggest that O. vulgatum extract prevents the skin cell damage induced by UV irradiation, and implies that O. vulgatum extract may be useful as a new ingredient for anti-aging cosmetics.

• Tuberculosis and Respiratory Diseases
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• v.57 no.5
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• pp.449-460
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• 2004

Background : PS-341 is a novel, highly selective and potent proteasome inhibitor, which showed cytotoxicity against some tumor cells. Its anti-tumor activity has been suggested to be associated with modulation of the expression of apoptosis-associated proteins, such as p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax and Bcl-2. c-Jun N-terminal kinase (JNK) and glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) are important modulators of apoptosis. However, their role in PS-341-induced apoptosis is unclear. This study was undertaken to elucidate the role of JNK and GSK-$3{\beta}$ in the PS-341-induced apoptosis in lung cancer cells. Method : NCI-H157 and A549 cells were used in the experiments. The cell viability was assayed using the MTT assay and apoptosis was evaluated by proteolysis of PARP. The JNK activity was measured by an in vitro immuno complex kinase assay and by phosphorylation of endogenous c-Jun. The protein expression was evaluated by Western blot analysis. Dominant negative JNK1 (DN-JNK1) and GSK-$3{\beta}$ were overexpressed using plasmid and adenovirus vectors, respectively. Result : PS-341 reduced the cell viability via apoptosis, activated JNK and increased the c-Jun expression. Blocking of the JNK activation by overexpression of DN-JNK1, or pretreatment with SP600125, suppressed the apoptosis induced by PS-341. The activation of caspase 3 was mediated by JNK activation. Blocking of the caspase 3 activation suppressed PS-341-induced apoptosis. PS-341 activated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, but its blockade enhanced the PS-341-induced cell death via apoptosis. GSK-$3{\beta}$ was inactivated by PS-341 via the PI3K/Akt pathway. Overexpression of constitutively active GSK-$3{\beta}$ enhanced PS-341-induced apoptosis; in contrast, this was suppressed by dominant negative GSK-$3{\beta}$ (DN-GSK-$3{\beta}$). Inactivation of GSK-$3{\beta}$ by pretreatment with lithium chloride or the overexpression of DN-GSK-$3{\beta}$ suppressed both the JNK activation and c-Jun up-regulation induced by PS-341. Conclusion : The JNK/caspase pathway is involved in PS-341-induced apoptosis, which is negatively regulated by the PI3K/Akt-mediated inactivation of GSK-$3{\beta}$ in lung cancer cells.