• 한방안이비인후피부과학회지
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• 제23권1호
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• pp.94-110
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• 2010

Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as phorbol-myristate-acetate(PMA), lipopolysaccharide(LPS), dermato-phagoides pteronyssus crude extract(DPE), dinitrochloro-benzene(DNCB) and Cryptotympana pustulata(CP), the Cryptotympana pustulata extracting substance at simultaneously on the translocation of nuclear factor-kappa B(NF-${\kappa}B$) towards to the nucleus and the mRNA expression patterns of various cytokine genes in Human acute monocytic leukemia cell line(THP-1 cells), monocytes of human. Experiment : To analyze cytokine genes expression patterns, the RT-PCR method was used, measuring tumor necrosis factor(TNF)-$\alpha$ that had been secreted during cell culture in the ELISA method. The morphological change in the cell observed during THP-1 cell culture was observed using a scanning electron microscope (SEM) and the quantitative distribution in the cell NF-${\kappa}B$ was analyzed through immunocytochemistry and a confocal microscopy. Result : CP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. The expressions of inter-leukin(IL)-10, interferon(INF)-$\gamma$, TNF-$\alpha$ and monocyte chemoattractantant protein(MCP)-1 induced by PMA were suppressed by CP while the expression of transforming growth factor(TGF)-$\beta$ was promoted. Regarding the secretion pattern of TNF-$\alpha$ according to PMA processing, its secretion amount was increased by CP concurrent processing, in case of processing CP onto PMA and LPS, We discovered that the secretion amount of TNF-$\alpha$ was increased. Upon processing PMA and LPS on the THP-1 cell strain at the same time or either additionally processing CP thereon, the movement increase towards the nucleus from the NF-${\kappa}B$ cell cytoplasm, a transcription factor was able to be observed. Conclusion : In this study, Cryptotympana pustulata extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances processed on the monocyte THP-1 cell of humans. Therefore, additional studies as for the immune adjusting function of Cryptotympana pustulata are considered to be able to offer important materials for curing immune abnormal diseases such as atopy dermatitis afterward.

• 한국식품영양학회지
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• 제23권3호
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• pp.306-310
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• 2010

This study was designed to investigate the protective effect of the combination of fucoidan and lutein against AAPH-induced oxidative stress in THP-1 cells. The combination of fucoidan and lutein existed significant antioxidant effect on AAPH-damaged THP-1 cells by using lipid peroxidation and cellular antioxidant capacity assay. Fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) did not affect at all the viability of THP-1 cells, but protected the AAPH-damage of THP-1 cells at the same concentration. The viability of THP-1 cells was 0% with 1 mM AAPH alone, the protective effect of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) was 37% and 36%, respectively. The combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) exhibited significant inhibitory effect of lipid peroxidation using TBARS assay and cellular antioxidant capacity using DCFH-DA assay. In lipid peroxidation, the TBARS value of 1 mM AAPH alone was $0.8{\pm}0.03\;nM$ MDA, its of the combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) was $0.2{\pm}0.05\;nM$ MDA. In cellular antioxidant capacity, the combination of fucoidan($1\;{\mu}g/m{\ell}$) and lutein($10\;{\mu}g/m{\ell}$) exhibited significant cellular antioxidant capacity of 76%, whereas quercetin($10\;{\mu}M$) as positive control exhibited the cellular antioxidant capacity of 32%. These results indicate that the cotreatment of fucoidan and lutein protects against AAPH-induced THP-1 cell damage by inhibiting lipid peroxidation, increasing cellular antioxidant capacity.

• 대한한의학회지
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• 제40권2호
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• pp.35-50
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• 2019

Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells. Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts ($0-300{\mu}g/ml$) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3 II, Beclin-1, Atg5, $NF-{\kappa}B$, p38, ERK was evaluated using western blot analysis. Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3 II protein expressions was increased. Activation of $NF-{\kappa}B$ was decreased and phosphorylation of p38 was increased. Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via $NF-{\kappa}B$ and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.

• IMMUNE NETWORK
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• 제10권5호
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• pp.173-180
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• 2010

Background: APRIL, originally known as a cytokine involved in B cell survival, is now known to regulate the inflammatory activation of macrophages. Although the signal initiated from APRIL has been demonstrated, its role in cellular activation is still not clear due to the presence of BAFF, a closely related member of TNF superfamily, which share same receptors (TACI and BCMA) with APRIL. Methods: Through transfection of siRNA, BAFF-deficient THP-1 cells (human macrophage-like cells) were generated and APRIL-mediated inflammatory activities were tested. The expression patterns of APRIL were also tested in vivo. Results: BAFF-deficient THP-1 cells responded to APRIL-stimulating agents such as monoclonal antibody against APRIL and soluble form of TACI or BCMA. Furthermore, co-incubation of the siBAFF-deficient THP-1 cells with a human B cell line (Ramos) resulted in an activation of THP-1 cells which was dependent on interactions between APRIL and TACI/BCMA. Immunohistochemical analysis of human pathologic samples detected the expression of both APRIL and TACI in macrophage-rich areas. Additionally, human macrophage primary culture expressed APRIL on the cell surface. Conclusion: These observations indicate that APRIL, which is expressed on macrophages in pathologic tissues with chronic inflammation, may mediate activation signals through its interaction with its counterparts via cell-to-cell interaction.

• 한방재활의학과학회지
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• 제19권2호
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• pp.89-102
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• 2009

Objectives : This study was carried out to understand the immunity responses and anti-oxidation effect of the Gamidaeganghwal-tang(GDT) on rheumatoid arthritis by using the THP-1 cells and the serum of CIA mice. Methods : For this purpose, GDT was orally administerd to mice with rheumatoid arthritis induced by collagen II. To investigate the immunity responses, value of cytokine and gene expression in the THP-1 cell, levels of cytokines in the serum of CIA(collagen type II induced arthritis) mice, number of immunocyte in PBMC of CIA mice were measured. Then, anti-oxidant activity, scavenging activity on DHHP(2,2-diphenyl-1-picrylhydrazyl) free radical and SOD(Superoxide dismutae)-like activity of GDT was observed. Results : 1. The levels of IL-$1{\beta}$, IL-6, IL-8, MCP-1 at 100, $50{\mu}g/m{\ell}$ of GDT were significantly reduced in the THP-1 cell. 2. The levels of TNF-${\alpha}$, COX-2 mRNA expression at 100, $50{\mu}g/m{\ell}$ of GDT and IL-$1{\beta}$, IL-6 at $100{\mu}g/m{\ell}$ of GDT were significantly reduced in the THP-1 cell line. 3. The levels of IL-6, TNF-${\alpha}$ and IL-$1{\beta}$ were significantly reduced in the serum of CIA mice. 4. The absolute number of CD3+, CD4+ and CD8+ cells were significantly induced, CD3+/CD69+, CD3+/CD49+, CD19+, B220+/CD23+ cells were significantly reduced in PBMC. 5. Scavenging activity on DPPH free radical and SOD-like activity were significantly induced in a concentration dependent manner. Conclusions : Taking all these observations, GDT considered to be effective in treating rheumatoid arthritis. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.

• IMMUNE NETWORK
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• 제4권2호
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• pp.116-122
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• 2004

Background: LIGHT (TNFSF14) is a member of tumor necrosis factor superfamily and is the ligand for TR2 (TNFRSF14/HVEM). LIGHT is known to have proinflammatory roles in atherosclerosis. Methods: To find out the expression pattern of LIGHT in atherosclerotic plaques, immunohistochemical analysis was performed on human carotid atherosclerotic plaque specimens. LIGHT induced atherogenic events using human monocytic cell line THP-1 were also investigated. Results: Immunohistochemical analysis revealed expression of LIGHT and TR2 in foam cell rich regions in the atherosclerotic plaques. Double immunohistochemical analysis further confirmed the expression of LIGHT in foam cells. Stimulation of THP-1 cells, which express TR2, with either recombinant LIGHT or immobilized anti-TR2 monoclonal antibody induced interleukin-8 and matrix metalloproteinase(MMP)-9. Electrophoretic mobility shift assay demonstrated that LIGHT induces nuclear localization of transcription factor, nuclear factor $(NF)-{\kappa}B$. LIGHT induced activation of MMP-9 is mediated by $NF-{\kappa}B$, since treatment of THP-1 cells with the $NF-{\kappa}B$ inhibitor PDTC (pyrrolidine dithiocarbamate) completely blocked the activation of MMP-9. Conclusion: These data indicate that LIGHT is expressed in foam cells in atherosclerotic plaques and is involved in atherogenesis through activation of pro-atherogenic cytokine IL-8 and destabilization of plaque by inducing matrix degrading enzyme.

• 생명과학회지
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• 제10권4호
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• pp.362-364
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• 2000

Opuntia ficus-indca(Op) extract has been claimed to have several therapeutic properties in oriental medicine including anti-inflammatory and anti-rheumatoid arthritis effects. Little is known of its effect on the activation of immune cells such as T cells and macrophages. To evaluate the functional effect of Op extract on immune cells, we examined whether Op extract stimulates the proliferation of T cells and the secretion of cytokines including IL-1 beta, IL-6 and tumor necrosis factor-alpha in THP-1 cell lines by RT-PCR. Op extract significantly enhanced the proliferation of T cell clone(D10S). Transcription of cytokines including IL-1 beta, IL-6, and TNF-alpha peaked 6 hrs after exposure to Op extract(100g/ml) in the THP-1 cell line and declined and declined thereafter. In an experiment to test the dose dependency of transcription of cytokines, transcription increased at a dose of 10 g/ml and the maximum expression was obtained at 100 g/ml, 6 hrs after exposure to Op extract. These findings suggest that Op extract is a potent stimulant of immune cells including T cells and macrophages, which acts by stimulating T cell proliferation and upregulating cytokines. These phenomena imply that some edible plants may be beneficial to living animals through the activation of immune functions.

• 동의생리병리학회지
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• 제20권1호
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• pp.43-51
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• 2006

This experiment was designed to investigate the effect of Amomum villosum(AMV) on the Alzheimer's disease. The effects of AMV extract on amyloid precursor proteins(APP), acetylcholinesterase(AChE), glial fibrillary acidic protein(GFAP) mRNA of PC-12 cell line treated by amyloid $\beta$ protein($A{\beta}$) : IL-$1{\beta}$, IL-6, TNF-$\alpha$ mRNA of THP-1 cell line treated by lipopolysaccharide(LPS) : AChE activity of PC-12 cell lysate treated by $A{\beta}$ : serum glucose, uric acid, AChE activity of memory deficit rats induced by scopolamine : behavior of memory deficit mice induced by scopolamine were investigated, respectively. AMV extract suppressed APP, AChE, GFAP mRNA in PC-12 cell treated by $A{\beta}$ : IL-$1{\beta}$, IL-6, TNF-$\alpha$ mRNA in THP-1 cell treated by LPS , AChE activity in cell lysate of PC-12 cell treated by $A{\beta}$. AMV extract increased glucose, decreased uric acid and AChE significantly in the serum of the memory deficit rats induced by scopolamine. AMV extract group showed significantly inhibitory effect on the memory deficit of mice induced by scopolamine in the experiment of Morris water maze. According to the above results, it is suggested that AMV extract might be usefully applied for prevention and treatment of Alzheimer's disease.

• International Journal of Oral Biology
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• 제42권1호
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• pp.33-38
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• 2017

Background: Periodontitis is an inflammatory disease characterized by the breakdown of tooth-supporting tissues, leading to tooth loss. Aggregatibacter actinomycetemcomitans are major etiologic bacterium causing aggressive periodontitis. Ursodeoxycholic acid (UDCA), a hydrophilic gall bladder acid, has been used as an effective drug for various diseases related to immunity. The aim of this study was to investigate the effect of UDCA on the inflammatory response induced by A. actinomycetemcomitans. Methods: A human acute monocytic leukemia cell line (THP-1) was differentiated to macrophage- like cells by treatment with phorbol 12-mystristate 13-acetate (PMA) and used for all experiments. The cytotoxic effect of UDCA was examined by MTT assay. THP-1 cells were pretreated with UDCA for 30 min before A. actinomycetemcomitans infection and the culture supernatant was analyzed for various cytokine production by ELISA. The effect of UDCA on bacterial growth was examined by measuring optical densities using a spectrophotometer. Results: UDCA showed no cytotoxic effect on THP-1 cells, up to $80{\mu}M$ Ed highlight: Please confirm technical meaning. UDCA pretreatment inhibited the A. actinomycetemcomitans-induced $IL-1{\beta}$, $TNF-{\alpha}$, and IL-17A secretion in a dose-dependent manner. UDCA also inhibited IL-21 production at $60{\mu}M$. The production of IL-12 and IL-4 was not influenced by A. actinomycetemcomitans infection. Conclusion: These findings indicate that UDCA inhibits the production of inflammatory cytokines involved in innate and Th17 immune responses in A. actinomycetemcomitans-infected THP-1- derived macrophages, which suggests its possible use for the control of aggressive periodontitis.

• 생명과학회지
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• 제34권4호
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• pp.271-278
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• 2024

Redox factor (Ref)-1은 세포질과 핵을 오가며 산화환원(redox) 환경에 민감한 transcription factors의 조절과 손상된 DNA의 교정 등 다양한 기능을 수행하는 단백질이다. 하지만, 단핵구(monocyte)의 대식세포(macrophage)로의 분화과정에서 Ref-1의 역할은 잘 알려져 있지 않다. 본 연구에서는 인간 단핵구세포주 THP-1을 이용하여 Ref-1의 단핵구 분화과정에 미치는 영향을 조사하였다. 분화제 phorbol 12-myristate 13-acetate (PMA)는 시간이 지날수록 세포의 부착능을 증가시키고 포식기능의 현저한 증가를 보이지만, Ref-1의 세포 내 양을 현저히 감소시켰다. Ref-1의 억제제인 E3330와 siRNA 기법을 이용한 Ref-1 knock- down은 PMA에 의한 세포 부착능과 막표면 분화인자의 발현을 현저히 감소시켰다. 이는 PMA에 자극을 받은 THP-1 세포의 분화 초기과정에는 Ref-1의 역할이 절대적으로 필요하다는 것을 의미한다. 단핵구 분화과정에서 Ref-1의 작용기전을 조사하기 위하여, PMA로 자극한 THP-1 세포의 세포질과 핵에서 Ref-1의 분포를 조사하였다. 놀랍게도, PMA 자극은 Ref-1을 빠르게 핵으로 이동하는 결과를 나타내었다. Ref-1의 핵으로의 이동이 단핵구 분화에 필요함을 증명하기 위하여, nuclear localization sequence (NLS)가 제거된 Ref-1 vector를 사용하였다. 그 결과, 핵으로의 이동이 제한된 ∆NLS Ref-1의 과발현은 PMA 자극에 의한 막표면 단백질의 발현 억제와 포식기능의 현저한 감소를 보였다. 이를 종합하면, Ref-1은 분화제 자극에 의한 분화 유도 초기과정에 핵으로 이동하여 다양한 분화인자의 발현에 관여하는 것으로 보인다.