• Nutrition Research and Practice
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• 제1권4호
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• pp.285-290
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• 2007

The leukocyte recruitment and transmigration across the endothelial barrier into the vessel wall are crucial steps in atherosclerosis. Leukocyte trafficking on the endothelium is elicited by induction of endothelial adhesion molecules, and its transmigration is mediated by degradation of basement membrane proteins through enzymatic activity of matrix metalloproteinases (MMP). The current study investigated whether resveratrol, a polyphenol present in grapes and red wine, was capable of inhibiting leukocyte adhesion to tumor necrosis factor (TNF)-${\alpha}$-activated endothelium. It was found that resveratrol inhibited the TNF-${\alpha}$-activated endothelial expression of vascular cell adhesion molecule-1 in a dose-dependent manner. In addition, resveratrol hampered THP-1 monocyte adhesion to activated endothelial cells. This study further examined whether resveratrol interfered with transendothelial migration of leukocytes. The MMP-2 gelatinolytic activity of endothelial cells was enhanced by TNF-${\alpha}$, which was attenuated by an addition of ${\geq}25{\mu}M$ resveratrol. In addition, 25 ${\mu}M$ resveratrol mitigated the MMP-9 activity of THP-1 cells, followed by a marked inhibition of transendothelial migration. These results demonstrated that resveratrol suppressed monocyte adhesion and migration induced by TNF-${\alpha}$ through modulating expression of adhesion molecules and gelatinolytic activity of MMP. These findings suggest that dietary resveratrol may be therapeutic agent for inhibiting leukocyte recruitment into the subendothelium during inflammatory atherosclerosis.

• 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.

• 한방안이비인후피부과학회지
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• 제23권2호
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• pp.57-68
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• 2010

Objective : This study was performed to evaluate the effect of Lithospermum erythrorhizon extracts on the inflammatory cytokines gene expression by the bacteria of Propionibacterium acnes (P. acnes) which elicits acne in human monocytes, THP-1 cell line. Experiment : Cytotoxicity of Lithospermum erythrorhizon extracts was analyzed by XTT assay. Real time RT-PCR was applied to analyze the cytokines gene expressions of IL-8, MCP-1 and TNF-$\alpha$. Translocation of transcription factor NF-${\kappa}B$ from cytoplasm into nucleus was observed using immunocytochemistry and confocal microscopy. Results : Lithospermum erythrorhizon extracts did not show cytotoxicity as high as in $1,000\;{\mu}g/ml$ of concentration. Transcription levels of inflammatory cytokines, IL-8, MCP-1 and TNF-$\alpha$ were increased by P. acnes in THP-1 and Lithospermum erythrorhizon extracts decreased the upregulated transcription levels. Lithospermum erythrorhizon extracts significantly inhibited the translocation of NF-${\kappa}B$ into nucleus by P. acnes. Conclusion : This study suggests that Lithospermum erythrorhizon extracts have anti-inflammatory effects on P. acnes treated THP-1 as decreasing the mRNA expressions of IL-8, MCP-1 and TNF-$\alpha$. This anti-inflammatory effect of Lithospermum erythrorhizon extracts may be useful in therapeutic treatments for acne vulgaris.

• 생명과학회지
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• 제29권11호
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• pp.1251-1257
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• 2019

본 연구에서는 펩티도글리칸이 단핵세포의 $TNF-{\alpha}$ 발현에 미치는 영향을 조사하였고, 또한 펩티도글리칸에 의한 $TNF-{\alpha}$ 발현에 관련된 세포의 요소들을 연구하였다. 사람의 단핵세포주인 THP-1 세포를 펩티도글리칸에 노출시키는 경우 $TNF-{\alpha}$ 분비 증가뿐만 아니라 $TNF-{\alpha}$ 유전자 전사를 유도하는 결과를 가져왔다. TLR-2/4의 억제제인 OxPAPC은 펩티도글리칸에 의한 $TNF-{\alpha}$의 발현을 저해하였다. 그리고 U0126, SB202190, SP6001250, LY294002, Akti IV, rapamycin, NAC, DPI 같은 약리학적 저해제 또한 $TNF-{\alpha}$ 발현을 유전자/단백질 수준에서 상당히 약화시켰다. 그러나 polymyxin B는 $TNF-{\alpha}$ 발현에 영향을 주지않았다. 따라서 펩티도글리칸이 TLR-2, PI3K, Akt, mTOR, MAPKs, ROS 등을 통하여 단핵세포의 $TNF-{\alpha}$ 발현을 증가시킴을 확인하였다.

• 한방재활의학과학회지
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• 제19권4호
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• pp.1-18
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• 2009

Objectives : This study was carried out to understand the immune responses of the Changchuldointanggami-bang(after referred to CDIT) on antioxidants, THP-1 cell and rheumatoid arthritis in Collagen-induced Arthritis(CIA) mice. Methods : The antioxidative effect of CDIT on DPPH free radical and SOD-like activity was measured. CDIT was administered to THP-1 cell, cytokine and mRNA associated with inflammation were measured. CDIT was orally administered to mice with arthritis by collagen II and then cytokine, PBMC in the serum were measured. Results : 1. The scavenging activity of CDIT on DPPH free radical was dose-dependent. 2. The effect of CDIT on SOD-like activity was dose-dependent. 3. The production of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ was decreased significantly in THP-1 cell. 4. The expression of $IL-1{\beta}$ mRNA, IL-6 mRNA, $TNF-{\alpha}$ mRNA were decreased significantly in THP-1 cell. 5. $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ were decreased significantly in the serum of CIA mice. 6. CD3+, CD4+, CD8+ were increased significantly, but CD3+/CD69+, CD3+/CD49b+, B220+/CD23+ were decreased significantly in PBMC of CIA mice. Conclusions : Taking all these observations into account, CDIT is considered to be effective in rheumatoid arthritis. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.

• International Journal of Oral Biology
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• 제42권3호
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• pp.117-121
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• 2017

We have previously shown that the specific phosphatidylinositol 3-kinase inhibitor LY294002 (LY29), and its inactive analog LY303511 (LY30), inhibit a monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells; these results suggest the potential of LY30 as an anti-inflammatory drug. In this study, we determined the effects of LY30 on the production of various inflammatory cytokines in human macrophagic THP-1 cells which were stimulated with lipopolysaccharide (LPS). LY30 selectively suppressed the mRNA expression of IL-12 p40, $TNF-{\alpha}$, and MCP-1 without affecting the expression of $IL-1{\alpha}$, IL-6, and IL-8. Inhibition of the production of IL-12 and $TNF-{\alpha}$ by LY30 was also demonstrated using ELISA assays. In order to elucidate the mechanisms of the action of LY30, we examined the role played by the mitogen-activated protein kinases and the key transcription factors, AP-1 and $NF-{\kappa}B$ in LPS-stimulated THP-1 cells. The results revealed that LY30 inhibited LPS-induced activation of ERK, but not p38 or JNK. Furthermore, the AP-1 DNA binding activity was suppressed by LY30 based upon the dosage, whereas $NF-{\kappa}B$ DNA binding was not affected. These results suggest that LY30 selectively inhibits cytokine production in the LPS-stimulated macrophagic THP-1 cells by down-regulating the activation of ERK and AP-1.

• 한국식품영양과학회지
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• 제31권6호
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• pp.1134-1141
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• 2002

염증성 cytokines의 분비 또는 혈관손상으로 인한 백혈구의 adhesion과 transmigration을 통하여 죽상경화과정이 시발되는데, 본 연구에서는 이러한 죽상경화의 초기과정에서 플라보노이드가 억제작용을 발휘하는 지를 규명하고자 하였다. 본 연구에서는 화학적인 구조가 서로 다른 플라보노이드를 사용하여 화학적인 구조와 항동맥경화작용과의 상관성을 착인하였다. TNF-$\alpha$는 혈관내피세포를 활성화시켜 THP-1 단핵구의 adhesion을 유의적으로 증가시켰다. 여러형태의 플라보노이드를 전처리하고 TNF-$\alpha$를 가하여 혈관내피세포를 활성화 시 켰을 때, flavonols인 quercetin과 flavones의 luteolin과 apigenin은 THP-1 단핵구의 adhesion억제효과를 보여주었다. 그러나, catechins과 flavanones의 플라보노이드는 이러한 억제효과를 전혀 보여주지 못하였다. 이러한 adhesion 억제작용을 가지는 플라보노이드는 CAMs 단백질의 발현도 차단시킨다는 것을 확인할 수 있었다. Quercetin, luteolin과 apigenin은 TNF-$\alpha$에 의하여 증가된 VCAM-1, ICAM-1 및 E-selectin의 단백질 발현을 일률적으로 감소 또는 차단시켰다. 그 대신, 단핵구의 adhesion을 차단시키지 못한 (-)epigallo-catechin gallate와 (+)catechin은 TNF-$\alpha$에 의한 이러한 CAMs의 발현을 전혀 억제시키지 못하였다. 또한 quercetin, luteolin과 apigenin의 CAMs단백질 발현 억제작용은 유전자 전사단계에서 mRNA의 down-regulation으로 인하여 나타난다는 사실을 알 수 있었다. 결론적으로 quercetin, luteolin, apigenin과 같은 플라보노이드는 TNF-$\alpha$와 같은 염증성 cytokines에 의한 단핵구의 adhesion을 혈관내피세포의 CAMs 단백질 발현을 억제하므로서 차단시킨다는 것이 확인되었다. 여기서 모든 플라보노이드가 이러한 활성을 다 지니고 있지 않아서 화학적인 구조와 초기 항동맥경화작용에는 서로 연관성이 있다는 것이 제시되었다. 또한, 선별된 플라보노이드의 초기 항동맥경화작용은 활성산소를 소거하는 플라보노이드의 항산화능과는 무관한 것 같다고 할 수 있다.

• IMMUNE NETWORK
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• 제7권1호
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• pp.26-30
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• 2007

Background: Mycobacterial antigens released as PIM, LM, LAM, lipoproteins and other cellular factors may contribute to macrophage and dendritic cell activation through pattern recognition receptors such as TLRs. In this study, we assessed cytokine production and ERK activation with stimulation of several major mycobacterial antigens. Methods: Purified mycobacterial antigens (10, 22, 30, 38kDa) and recombinant antigens (6, 16, 19, 38kDa, Ag85A antigen) were studied. The production of cytokines (TNF-${\alpha}$, IL-12, IL-6) was measured by ELISA. The ERK activation was detected by western blotting. The expression of TLR2 or TLR4 was measured by flow cytometry. Results: Among purified antigens only 30kDa antigen induced production of IL-6 or TNF-${\alpha}$ in THP-1 macrophage cells. When THP-1 macrophage cells were treated with 30kDa antigen, phosphorylation of ERK was detected. ERK activation also occurred in TLR2 transfectant HEK293 cells with 30kDa antigen stimulation. Conclusion: 30kDa antigen is one of the major mycobacterial antigens inducing cytokine production and MAP kinases phosphorylation in macrophages.

• Journal of Applied Biological Chemistry
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• 제61권2호
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• pp.181-186
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• 2018

본 연구에서는 인간 단핵구 세포인 THP-1 세포를 이용하여 berberine의 항염증 활성을 조사 하였다. Propionibacterium acnes의 감염은 THP-1 세포에서 산화질소(NO)와 $TNF-{\alpha}$, IL-8 및 $IL-1{\beta}$와 같은 전 염증성 사이토카인의 생산을 유도했다. 그러나, P. acnes에 의해 유도된 THP-1 세포에 berberine을 처리했을 때, 전 염증성 사이토카인 및 NO의 생성이 유의하게 감소하였다. 또한 우리는 berberine의 항 염증 기능의 신호 전달 경로를 분석하여 berberine이 P. acnes 유도 세포에서 ERK1/2, JNK 및 p38의 인산화를 억제하고 $NF-{\kappa}B$ p65의 발현 및 핵이동을 억제한다는 것을 발견했다. 이러한 결과로부터 berberine은 인간 단핵구 세포에서 $NF-{\kappa}B$ 및 MAPK 신호 전달 경로를 억제함으로써 항 염증 활성을 효과적으로 발휘할 수 있다고 결론지었다. 또한, 이러한 결과는 P. acnes에 의해 유발된 염증성 질환의 치료를 위해 천연물 소재에서 유래한 알칼로이드 화합물인 berberine을 사용하여 천연 치료제를 개발할 수 있는 가능성을 제시하였다.

• 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.