• Tuberculosis and Respiratory Diseases
• /
• 제52권2호
• /
• pp.145-155
• /
• 2002

연구배경 : 혈관내피세포와 호중구의 상호작용은 급성염증반응의 생리 및 병리학적 핵심적인 과정으로서 특별한 분자들에 의해서 매개되고 있다. 적절한 자극이 주어질 때 혈관 내피세포에서는 부착 및 신호인자가 표현되며 이들은 호중구의 수용체에 의해 인지된다. 본 연구에서는 IL-1과 LPS로 혈관내피세포를 자극하였을 때 CXC chemokine family에 속하는 GRO-${\alpha}$, IL-8 및 ENA-78이 내피세포에서 분비되는 양상을 관찰하였고, ENA-78과 GRO-${\alpha}$가 호중구-내피세포의 부착에 미치는 영향을 평가하였다. 방 법 : 인제대혈관 내피세포를 배양하여 다양한 농도로 IL-1과 LPS로 자극하였다. 자극된 내피세로로부터 분비 되어진 GRO-${\alpha}$, IL-8 및 ENA-78의 농도는 ELISA로 측정하였다. recombinant human ENA-78 및 GRO-${\alpha}$로 자극된 내피세포에 동위원소가 부착된 정상백혈구를 이용하여 부착능을 측정하였다. 결 과 : IL-1a과 LPS로 자극된 내피세포에서 GRO-${\alpha}$ IL-8 및 ENA-78의 분비는 자극의 농도와 지속시간에 비례하여 증가되었고, recombinant human ENA-78과 GRO-${\alpha}$의 농도가 증가함에 따라 호중구의 부착율이 더욱 증가하였다. 결 론 : ENA-78과 GRO-${\alpha}$는 CXC chemokine family에 속하며 주로 상피세포에서 발현되어 백혈구의 화학적 주성을 유발하며 염증성 반응을 매개하는 중요한 물질로 알려져 있다. 본 연구에서는 다양한 농도의 IL-1과 LPS로 자극된 내피세포에서 농도에 비례하여 GRO-${\alpha}$, IL-8 및 ENA-78이 분비됨을 확인할 수 있었으며, 또한 ENA-78과 GRO-${\alpha}$를 각각 다른 농도로 배양한 후 호중구 부착능이 농도에 따라 증가함을 알 수 있었다. 따라서 본 연구의 결과는 염증성 자극으로 혈관내피세포에서 분비되는 ENA-78과 GRO-${\alpha}$ 등의 CXC chemokine 이 염증반응의 진행 기전에서 중요한 역할을 담당할 것임을 시사하며, 향후 염증성 질환의 발병기전 규명이나 치료법개발을 위한 기초적 자료를 제공하는데 도움이 될 것으로 생각된다.

• 동의생리병리학회지
• /
• 제21권1호
• /
• pp.39-49
• /
• 2007

Rheumatoid arthritis is an autoimmune disease involving multiple joint. In order to access the suppressive effects of JTT on rheumatoid arthritis and it's effects on immune system we investigated whether JTT could suppress the disease progression of collagen-induced arthritis. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with DW, JTT (200 or 400 mg/kg) or methotrexate (MTX, 30 mg/kg) as a positive control. Oral administration of JTT significantly suppressed the progression of CIA, which extend is comparable to that of MTX. Histological examination reveled that JTT inhibited infiltration of inflammatory cells into affected paw joint and bone erosion and cartilage destruction were greatly reduced compared with control. Total cell number of spleen, lymph node and peripheral blood were significantly reduced. The absolute number of CD19$^+$, CD3$^+$/CD69$^+$, CD4$^+$/CD25$^+$ cell in spleen from JTT treated mice were significantly decreased. The absolute number of CD19$^+$, CD3$^+$, CD3$^+$/CD69$^+$, CD4$^+$, CD4$^+$/CD25$^+$ CD8$^+$, CD49b, CD3/CD49b cells in draining lymph node were significantly increased compared with control. In peripheral blood mononuclear cells of JTT treated mice, the absolute number of CD4$^+$, CD4$^+$/CD25$^+$, CD3$^+$/CD69$^+$ cells were significantly decreased compared with control, while that of CD49b$^+$ was slightly increased. Infiltration of CD3$^+$ cells and CD11b$^+$/Gr-1$^+$ cells into paw joint was significantly reduced in JTT treated mice. The levels of pathologic cytokines including TNF-a and IL-6 in serum were significantly decreased by oral treatment with JTT The levels of IFN-g in the culture supernatant of splenocyte stimulated with CD3$^+$/CD28$^+$ or collagen were dramatically decreased, while the levels of IL-4 was increased under CD3$^+$/CD28$^+$ or collagen stimulation. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of JTT treated mice than control. Taken together, JTT has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use anti-rheumatic arthritic agent in human.

• Journal of Acupuncture Research
• /
• 제27권4호
• /
• pp.39-53
• /
• 2010

목적 : 퇴행성 관절염은 염증성 사이토카인인 IL-$1\beta$에 의해 연골관절이 파괴되고 이로 인해 염증성 사이토카인이 더욱 증가하는 질환이다. 퇴행성 관절염을 치료하기 위해서는 연골 파괴를 가속화시키는 catabolic cytokines의 활성을 줄이고, 성장인자인 anabolic factor의 활성을 증가시는 연골 보호 작용이 있어야 한다. 본 연구에서는 독활 승마 처방(OAH19T)이 catabolic/anabolic 대사 조절에 어떤 영향을 미치는지와 그 신호 전달 기전에 대해 연구하였다. 또한 OAH19T를 구성하는 단미재 및 임상에서 사용되는 COX-2 inhibitor인 Celebrex(CEL)와 효능을 비교 실험하였다. 방법 : 배양된 세포에 IL-$1\beta$로 자극한 후 (1) glycosaminoglycan(GAG)의 분해 억제 정도, (2) OAH19T와 CEL에 대하여 MMP-1과 MMP-3의 유전자 발현 및 활성 억제, (3) Aggrecan 및 Aggrecanases의 유전자 발현 및 활성 억제, (4) OAH19T의 growth factor의 조절 능력, (5) MAPK pathway 등을 RT-PCR(reverse transcriptase-polymerase chain reaction), ELISA(Enzyme-linked immunosorbent assay), western blot, viability 측정을 통해 검증했다. 결과 : 사람 관절 세포에서 (1) 독활 승마 각각의 단미재, 임상에서 사용중인 셀레콕시브(CEL), 조인스보다 실험 약물(OAH19T)이 저농도에서 GAG 분해 억제 효과가 우수하였고, 부탄올로 분획한 OAH19B와는 동등한 효과를 보였다. (2) OAH19T는 IL-$1\beta$에 의하여 활성화된 MMP-1과 MMP-3의 발현을 모두 억제하였으나, CEL은 MMP-1의 발현은 억제하였으나 MMP-3의 발현은 억제하지 못하였다. (3) OAH19T는 IL-$1\beta$에 의하여 손상된 Aggrecan을 회복시켰으며 이는 활성화된 Aggrecanase-1과 Aggrecanase-2를 억제시킴으로써 나타난 결과이다. 그러나 CEL의 경우, 손상된 Aggrecan을 회복시키지 못하였다. (4) 배양된 세포는 IL-$1\beta$에 의하여 TGF-$\beta$II및 TGF-$\beta$ receptor II의 발현이 억제되었으나, OAH19T는 TGF-$\beta$II및 TGF-$\beta$ receptor II의 발현을 회복시켜 OAH19T가 anabolic한 조절능력이 있음을 시사한다. 그러나 CEL의 경우 growth factor에 대한 조절 능력이 없었다. (5) 대사 조절 작용에 대한 기전으로서 MAPK pathway에 대해서 연구한 결과 IL-$1\beta$에 의하여 유도된 pERK, pp38 kinase의 활성은 억제하였고, pJNK의 활성은 변하지 않았다. 또한 OAH19T는 연골 세포에 독성이 없었으며 IL-$1\beta$에 의해 유도된 세포 증식만을 억제시켰다. 이 결과로, OAH19T가 OA chondrocyte의 탈분화 및 세포 고사를 억제하여 연골보호 및 회복 효과가 있음을 알 수 있었다. 결론 : OAH19T는 이를 구성하는 단미재 및 CEL보다 연골보호 효과가 월등하였고, 이러한 연골보호 효과는 catabolic cytokines/growth factors의 균형으로 대사조절을 통해 연골세포의 탈분화 및 세포 고사를 억제하여 연골보호 및 회복 효과가 있음을 알 수 있었다.

• Tuberculosis and Respiratory Diseases
• /
• 제62권3호
• /
• pp.197-202
• /
• 2007

흡연물질로서 benzopyrene, nicotine 및 cotinine등은 기관지상피세포인 Beas2B에서 CuZnSOD, thioredoxin, glutathione reductase 등의 발현량에 영향을 주며 특히 thioredoxin 및 glutathione reductase의 발현량을 노출 후 30분에서 4시간 경과 시간대에 증가시켰다가 24시간 이후에는 억제하는 특징을 나타내었다. 상기한 흡연물질에 의한 항산화효소의 조절기전에는 전사인자인 NNF-${\kappa}B$가 관여하는 것으로 추정되었다.

• 생명과학회지
• /
• 제16권3호
• /
• pp.500-504
• /
• 2006

합토글로빈(Hp)은 조직손상이나 감염 등의 염증시에 혈중 농도가 증가하는 급성기반응단백질로서 주로 간에서 생합성되지만 염증을 동반한 폐에서도 발현됨이 보고되었다. 폐에서 합성되는 Hp이 염증반응에 어떤 영향을 주는지를 조사하고자, Hp을 과발현하는 폐상피세포를 사용하여 COX-2 및 염증관련 cytokine들의 발현을 조사하였다. Stable transfection 또는 transient transfection된 A549 세포에서 Hp이 잘 발현되고 있음을 확인한 후 COX-2의 발현을 측정한 결과, Hp의 과발현에 의해 COX-2 합성이 현저히 증가함을 확인할 수 있었다. 또한 Hp을 과발현하는 A549세포에 LPS ($1{\mu}g/ml$) 또는 $IL-1{\beta}$ (100 U/ml)를 각각 24시간 동안 처치하였을 때, Hp에 의한 COX-2 발현 증가는 LPS또는 $IL-1{\beta}$자극에 의해 협동적으로 증가하였다. ACP-based PCR 방법으로 염증관련 cytokine들의 발현을 측정한 결과, Hp에 의해 SPARC의 발현이 현저히 저하되는 반면에 IL-4 및 S100A1 유전자 발현은 약간 증가함을 알 수 있었다. 이러한 결과들은 폐상피세포에서 Hp이 염증반응을 활성화시키는 기전으로 작용함을 시사한다.

• Journal of Acupuncture Research
• /
• 제26권5호
• /
• pp.65-75
• /
• 2009

Objectives : Pyrite is one of the important prescriptions that has been used in oriental medicine for healing of fracture. It is reasonable, therefore, to postulate that native copper affects the process of bone metabolism and bone formation. The purpose of this study is to discover the effect of Pyrite on the healing of tibia fracture. Methods : 1. In vitro test : MG-63 cell in human body and the Pyritum in the ratio of 0.5mg/ml, 1.0mg/ml, 1.5mg/ml, 2.0mg/ml were incubated for 24 hours. After 24 hours, RNA was extracted via trizol reagent (Sigma, USA). In order to understand the activation of osteoblast, the level of OPN mRNA, osteopontin, was measured. 2. In vivo tesgroups normal group, control group and experimental group. Left tibia bones of mice in CON and JT groups were fractured by bone cutters. Pyrite was orally administered to the experimental group. After 14 days, each group's tibia specimen was constructed to observe changes in activation of proinflmmatory cytokines in relation to MIF and IL-6. Also, proliferation of osteoblast and osteopontin were measured via changes in levels of OPN and OPN mRNA. Results : In jn-Titro test, the level of OPN mRNA, osteopontin production was remarkably increased in Pyritum-treated MG-63 cells. In in-vitro test, fractured area in external tibia morphology was increased more in the JT group than that of the CON group. Osteogenesis, endochodrial ossification, and osteoid in fractured area were also increased more in the JT group than that of the CON group. Increase in OPN mRNA, osteopontin level and osteoblast's proliferation were observed. Activation of MIF and IL-6 was confirmed from the fracture region. Conclusions : From the result, development of a new stimulator in healing fracture via pyrite is expected.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제38권
• /
• pp.48.1-48.8
• /
• 2016

Background: This study investigates the effect of alendronate-treated osteoblasts, as well as the effect of low-level laser therapy (LLLT) on the alendronate-treated osteoblasts. Bisphosphonate decreases the osteoblastic activity. Various treatment modalities are used to enhance the bisphosphonate-treated osteoblasts; however, there were no cell culture studies conducted using a low-level laser. Methods: Human fetal osteoblastic (hFOB 1.19) cells were treated with $50{\mu}M$ alendronate. Then, they were irradiated with a $1.2J/cm^2$ low-level Ga-Al-As laser (${\lambda}=808{\pm}3nm$, 80 mW, and 80 mA; spot size, $1 cm^2$; NDLux, Seoul, Korea). The cell survivability was measured with the MTT assay. The three cytokines of osteoblasts, receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL), osteoprotegerin (OPG), and macrophage colony-stimulating factor (M-CSF) were analyzed. Results: In the cells treated with alendronate at concentrations of $50{\mu}M$ and higher, cell survivability significantly decreased after 48 h (p < 0.05). After the applications of low-level laser on alendronate-treated cells, cell survivability significantly increased at 72 h (p < 0.05). The expressions of OPG, RANKL, and M-CSF have decreased via the alendronate. The RANKL and M-CSF expressions have increased, but the OPG was not significantly affected by the LLLT. Conclusions: The LLLT does not affect the OPG expression in the hFOB cell line, but it may increase the RANKL and M-CSF expressions, thereby resulting in positive effects on osteoclastogenesis and bone remodeling.

• Tuberculosis and Respiratory Diseases
• /
• 제79권4호
• /
• pp.257-266
• /
• 2016

Background: Idiopathic pulmonary fibrosis is a common interstitial lung disease; it is a chronic, progressive, and fatal lung disease of unknown etiology. Over the last two decades, knowledge about the underlying mechanisms of pulmonary fibrosis has improved markedly and facilitated the identification of potential targets for novel therapies. However, despite the large number of antifibrotic drugs being described in experimental pre-clinical studies, the translation of these findings into clinical practices has not been accomplished yet. NADH:quinone oxidoreductase 1 (NQO1) is a homodimeric enzyme that catalyzes the oxidation of NADH to $NAD^+$ by various quinones and thereby elevates the intracellular $NAD^+$ levels. In this study, we examined the effect of increase in cellular $NAD^+$ levels on bleomycin-induced lung fibrosis in mice. Methods: C57BL/6 mice were treated with intratracheal instillation of bleomycin. The mice were orally administered with ${\beta}$-lapachone from 3 days before exposure to bleomycin to 1-3 weeks after exposure to bleomycin. Bronchoalveolar lavage fluid (BALF) was collected for analyzing the infiltration of immune cells. In vitro, A549 cells were treated with transforming growth factor ${\beta}1$ (TGF-${\beta}1$) and ${\beta}$-lapachone to analyze the extracellular matrix (ECM) and epithelial-mesenchymal transition (EMT). Results: ${\beta}$-Lapachone strongly attenuated bleomycin-induced lung inflammation and fibrosis, characterized by histological staining, infiltrated immune cells in BALF, inflammatory cytokines, fibrotic score, and TGF-${\beta}1$, ${\alpha}$-smooth muscle actin accumulation. In addition, ${\beta}$-lapachone showed a protective role in TGF-${\beta}1$-induced ECM expression and EMT in A549 cells. Conclusion: Our results suggest that ${\beta}$-lapachone can protect against bleomycin-induced lung inflammation and fibrosis in mice and TGF-${\beta}1$-induced EMT in vitro, by elevating the $NAD^+$/NADH ratio through NQO1 activation.

• Food Science and Biotechnology
• /
• 제17권5호
• /
• pp.953-958
• /
• 2008

The object of this study was to investigate the immune-enhancing effects of Chlorella vulgaris (CV) on a deteriorated immune function by a protein-energy malnutrition (PEM) diet. Unicellular algae, CV were used as a biological response modifier. Male C57BL/6J mice were fed for 15 days with standard diet or a PEM diet, which is associated with decreased host immune defense. After 8 days, mice in the PEM diet group were orally administered by 0.05, 0.1, and 0.15 g/kg body weight of CV or distilled water. Nutritional parameters, and interferon (IFN)-$\gamma$ levels were significantly increased in the blood serum of the CV (0.15 g/kg)-treated group (29.6$\pm$2.8 pg/mL) compared to the non-treated PEM group (4.1$\pm$0.4 pg/mL, p<0.05). In addition, cell proliferation and production of cytokines were investigated via a CV (0.01, 0.1, and 1 mg/mL) treatment using a human T cell line MOLT-4 cell. The CV treatment (1 mg/mL) significantly increased the production of both IFN-$\gamma$ and interleukin (IL)-2 (51.3$\pm$3.4 and 285.9$\pm$18.8 pg/mL, respectively) compared to the control (51.3$\pm$3.4 and 442.6$\pm$14.3 pg/mL, respectively), but did not affect the production of IL-4. These results suggest that CV may be useful in improving the immune function.

• Molecules and Cells
• /
• 제27권1호
• /
• pp.105-111
• /
• 2009

Gammaherpesvirus infection of the central nervous system (CNS) has been linked to various neurological diseases, including meningitis, encephalitis, and multiple sclerosis. However, little is known about the interactions between the virus and the CNS in vitro or in vivo. Murine gammaherpesvirus 68 (MHV-68 or ${\gamma}HV-68$) is genetically related and biologically similar to human gammaherpesviruses, thereby providing a tractable animal model system in which to study both viral pathogenesis and replication. In the present study, we show the successful infection of cultured neuronal cells, microglia, and astrocytes with MHV-68 to various extents. Upon intracerebroventricular injection of a recombinant virus (MHV-68/LacZ) into 4-5-week-old and 9-10-week-old mice, the 4-5-week-old mice displayed high mortality within 5-7 days, while the majority of the 9-10-week-old mice survived until the end of the experimental period. Until a peak at 3-4 days post-infection, viral DNA replication and gene expression were similar in the brains of both mouse groups, but only the 9-10-week-old mice were able to subdue viral DNA replication and gene expression after 5 days post-infection. Pro-inflammatory cytokine mRNAs of tumor necrosis factor-${\alpha}$, interleukin $1{\beta}$, and interleukin 6 were highly induced in the brains of the 4-5-week-old mice, suggesting their possible contributions as neurotoxic factors in the age-dependent control of MHV-68 replication of the CNS.