• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.645-654
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• 1999

Interleukin-6(IL-6) stimulate osteoclast differentiation. $17{\beta}$-estradiol, 1,25-dihydroxyvitamin $D_3$(1,25-$(OH)_2D_3$) and interleukin-$1{\beta}$ inhibit or stimulate osteoclast differentiation by decreasing or increasing the synthesis of interleukin-6(IL-6) from stromal/osteoblastic cells, respectively. Periodontal ligament(PDL) cells reside between the alveolar bone and the cementum and have osteoblastic characteristics. To estimate the effect of $17{\beta}$-estradiol and 1,25$(OH)_2D_3$ on IL-6 production of PDL cells, PDL cells were treated with $17{\beta}$-estradiol or 1,25-$(OH)_2D_3$ in the absence or the presence of IL-$1{\beta}$. The concentration of IL-6 produced form PDL cells was determined by enzym linked immunosorbent assay(ELISA). In unstimulated PDL cells, we detected constitutive production of IL-6 at 1st and 2nd day. IL-$1{\beta}$ increased IL-6 synthesis at 1st day and 2nd day. $17{\beta}$-estradiol had no significant effect on the secretion of this cytokine, either constitutively or after stimulation with IL- $1{\beta}$(0.05 ng/ml). 1,25-$(OH)_2D_3$($10^{-8}M$) decreased not only constitutive IL-6 production but also IL-$1{\beta}$-induced IL-6 production at 2nd day. These results suggest that 1,25-$(OH)_2D_3$ may control IL-$1{\beta}$-induced osteoclast differentiation by decreasing IL-$1{\beta}$-induced IL-6 secretion of PDL cells.

• Korean Journal of Acupuncture
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• v.27 no.2
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• pp.95-105
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• 2010

Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.3
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• pp.1-15
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• 2012

Objectives: The purpose of this study was to investigate the effects of Chaenomelis Fructus Herba Water Extract(CF) on the production of inflammatory mediators in RAW 264.7 cell mouse macrophages stimulated with LPS. Methods: We have not examined effect of CF on the cell viability of RAW 264.7 cell until we investigated effects of CF on LPS-induced productions of NO, Ca and various cytokines in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically(P<0.05). Results: 1. CF increased the cell viability in the RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 2. CF inhibited significantly increasing the production of NO in LPS-induced RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 3. CF inhibited significantly increasing the production of Intracellular Ca in LPS-induced RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 4. CF inhibited significantly the IL-2, IL-10, IL-12p70, TNF-${\alpha}$, GM-CSF, M-CSF, LIF and VEGF of the RAW 264.7 cell induced by LPS at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 5. CF inhibited significantly the IL-4 at the density of 25, 50 ${\mu}g/ml$, the IL-5, IL-15 and MIG at the density of 25, 50 and 200 ${\mu}g/ml$ and IFN-${\gamma}$ at the density of 25, 100 ${\mu}g/ml$ respectively in the RAW 264.7 cell increased by LPS. Conclusions: CF inhibited significantly increasing IL-2, IL-10, IL-12p70, TNF-${\alpha}$, GM-CSF, M-CSF, LIF, VEGF, NO and Ca in LPS-induced RAW 264.7 cell at the density of more than 25 ${\mu}g/ml$ without causing the toxicity. These results signify that CF has antiinflammatory effect on controlling the over inflammatory reaction by the RAW 264.7 cell.

• The Journal of Internal Korean Medicine
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• v.31 no.2
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• pp.189-200
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• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.795-801
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• 2013

The purpose of this study is to investigate effects of White Ginseng-Ejung-tang (EG) and Red Ginseng-Ejung-tang (ER) water extract on production of various cytokines such as interleukin (IL)-21, IL-25, IL-$28{\beta}$, erythropoietin (EPO), Exodus-2, monocyte chemotactic protein (MCP)-5, macrophage inflammatory protein (MIP)-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, and TARC in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. ER significantly decreased levels of IL-21, IL-25, IL-$28{\beta}$, EPO, Exodus-2, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, TARC, and fractalkine for 24 h incubation at the oncentrations of 25 and 100 ${\mu}g/mL$ in LPS-induced RAW 264.7 (P < 0.05). But EG did not show any significant effect. These results suggest that ER has anti-inflammtory property related with its inhibition on the production of IL-21, IL-25, IL-$28{\beta}$, and chemokines such as EPO, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, Exodus-2, and TARC in LPS-induced macrophages.

• Journal of Acupuncture Research
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• v.35 no.1
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• pp.11-20
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• 2018

Background: This research was performed to investigate the effects of Pulsatilla Koreana NAKAI pharmacopuncture (PPA) therapy on intestinal disease in rats with dextran sulfate sodium (DSS)-induced colitis. Methods: The subjects were divided into five groups : A control group, saline group, pharmacopuncture group PPA1 ($0.2mg/1kg/40{\mu}{\ell}$), pharmacopuncture group PPA2 ($0.5mg/1kg/40{\mu}{\ell}$), and pharmacopuncture group PPA 3($1mg/1kg/40{\mu}{\ell}$). The experimental model of colitis was induced by infection of dextran sulfate sodium (DSS) for eighteen days. After colitis was induced, PPA therapy was practiced on the Chunchu (ST25) once every two days for a total six times. Thereafter Disease Activity Index (DAI), colon length, damage to the colonic mucosa, body weight, IL-6, IL-10, $IL-1{\beta}$, $IFN-{\gamma}$, $TNF-{\alpha}$, $TGF-{\beta}1$, IL-23 and IL-17 were measured. Results: The results were as follows. 1. DAI was significantly decreased in the PPA groups. 2. Colon length was significantly increased in the PPA groups. 3. Damage of colonic mucosa was observed less in the PPA groups. 4. Body weight was significantly increased in the saline group and the PPA groups. 5. The PPA2 group showed a significant decrease in the intensity of IL-6, $IL-1{\beta}$, $IFN-{\gamma}$ and $TNF-{\alpha}$ levels and the mean of IL-23. 6. The PPA3 group showed a significant increase in the intensity of IL-10 and $TGF-{\beta}1$ levels. 7. No significant differences were shown in the mean of IL-17. Conclusion: These results suggest that PPA therapy on Chunchu (ST25) can be used as an effective treatment for inflammatory bowel disease.

• IMMUNE NETWORK
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• v.22 no.3
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• pp.25.1-25.11
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• 2022

IL-34 can promote osteoclast differentiation and activation, which may contribute to steroid-induced osteonecrosis of the femoral head (ONFH). Animal model was constructed in both BALB/c and IL-34 deficient mice to detect the relative expression of inflammation cytokines. Micro-CT was utilized to reveal the internal structure. In vitro differentiated osteoclast was induced by culturing bone marrow-derived macrophages with IL-34 conditioned medium or M-CSF. The relative expression of pro-inflammation cytokines, osteoclast marker genes, and relevant pathways molecules was detected with quantitative real-time RT-PCR, ELISA, and Western blot. Up-regulated IL-34 expression could be detected in the serum of ONFH patients and femoral heads of ONFH mice. IL-34 deficient mice showed the resistance to ONFH induction with the up-regulated trabecular number, trabecular thickness, bone value fraction, and down-regulated trabecular separation. On the other hand, inflammatory cytokines, such as TNF-α, IFN-γ, IL-6, IL-12, IL-2, and IL-17A, showed diminished expression in IL-34 deficient ONFH induced mice. IL-34 alone or works in coordination with M-CSF to promote osteoclastogenesis and activate ERK, STAT3, and non-canonical NF-κB pathways. These data demonstrate that IL-34 can promote the differentiation of osteoclast through ERK, STAT3, and non-canonical NF-κB pathways to aggravate steroid-induced ONFH, and IL-34 can be considered as a treatment target.

• 한국해양학회지
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• v.21 no.4
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• pp.245-258
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• 1986

A two-dimensional numerical model is developed to investigate the phenomenon of forced seiche caused by the incident long waves. This model is applied to Young-il Bay and Pohang New Harbor, where the seiche is most frequently observed and the damage of the seiche is serious compared with the other harbors in Korea. Some results on this study are as follows; 1. The natural periods of the first two modes obtained from the present model are about 70 and 25 minutes respectively for Young-il Bay, and about 25 and 7.5 minutes for Pohang New Harbor. These results are well consistent with those obtained by the theoretical formula, the spectral analysis, and the statistical investigation of the recorded data. 2. Since the fundamental natural period of Pohang New Harbor is almost the same as the second natural period of Young-il Bay, the seiche in Pohang New Harbor can be strongly amplified by the Oscillation in Young-il Bay. Therefore, the most strong seiche in Pohang New Harbor can occur when the long wave of about 25-minute period come into Young-il Bay.

• The Journal of Korean Obstetrics and Gynecology
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• v.15 no.4
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• pp.45-60
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• 2002

Mouse calvarial osteoblast cells were isolated and cultured. To examine whether the cells produce active gelatinases in culture medium or not,the cells were analyzed using by zymograsphic analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase- A. Then, mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agent's, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of $IL-1{\alpha}$, indomethacin and dexametasone produced a rightward shift in the IL-1 dose response curve. The results of in vitro cytotoxicities showed that Taeyoungjon-Jahage water extracts(T.Y.J-J.H.G extracts) have no any cytotoxicities in concentrations of $1-200\;{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $300\;{\mu}g/ml$ on mouse calvarial bone cells. T.Y.J.-J.H.G. extracts had protective activity against PTH (2 units/mI), or MCM (5%, v/v), or $rhIL-1{\alpha}$ (1 ng/mI) or $1,25(OH)_2D3$ (10 ng/ml) , $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the T.Y.J.-J.H.G.extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore. the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resortion agents of PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ and $IL-1{\alpha}$, with strong protective effect in pretreatment with the extracts. T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1 stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by T.Y.J.-J.H.G. extracts treatment in the mouse calvarial tissue culture system. These results indicated that the T.Y.J.-J.H.G.-water extracts are highly stable and applicable to clinical uses in osteoporosis.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.15-25
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• 2004

Objective : We studied the effect of Bambusae concretio Silicea (BCS) on bone metabolism. Methods : At first, we treated PTH, 1,25(OH)₂D₃, mononuclear cell conditioned medium (MCM) and IL-1 to osteoblast cells derived from mouse calvarial bone explants in vitro, and then investigated the activities of collagenolysis and bone resorption factors. Results : BCS extracts have no cytotoxicities in concentrations of 1-150 ㎍/ml. BCS had protective activity against PTH (5 units/ml), MCM (5%, v/v), 1,25(OH)₂D₃ (20 ng/ml), IL-1α(2 ng/ml) and IL-1β, (1 ng/ml)-induced collagenolysis in the mouse calvarial cells. And, pretreatment of BCS for 1 hr significantly reduced the collagenolysis. Furthermore, it was much more expressed at 16 hrs after BCS (50 ㎍/ml)-pretreatment. And, BCS significantly protected against enhanced collagenolysis induced by IL-1α and IL-1β. Conclusion : BCS extracts inhibited the bone resorption in mouse calvarial bone cell;, thus BCS could be used clinically for bone diseases.