• Tuberculosis and Respiratory Diseases
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• 제57권5호
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• pp.449-460
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• 2004

연구배경 : PS-341은 최근에 개발된 강력하고 특이적인 proteasome 억제제로서, 일부 암환자에 투여하여 좋은 성적이 보고되고 있다. Proteasome 억제제의 항암효과는 아포프토시스 유발 물질, 즉 p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax, Bcl-2 등의 발현 증가와 관련이 있는 것으로 생각되고 있다. JNK와 GSK-$3{\beta}$도 아포프토시스에 관여하는 것으로 잘 알려져 있지만, PS-341에 의한 아포프토시스에서의 역할은 규명되지 못한 상태이다. 본 연구에서는 폐암세포주에서 PS-341에 의한 아포프토시스에서 JNK와 GSK-$3{\beta}$의 역할을 규명하고자 하였다. 방 법 : NCI-H157과 A549 폐암세포주를 실험에 사용하였다. 세포생존능은 MTT 방법으로 평가하였고, 아포프토시스는 PARP의 분해로 평가하였다. JNK의 활성도는 in vitro immuno complex kinase 방법과 내인성 c-Jun의 인산화로 측정하였다. 각종 단백의 발현은 Western 분석으로 평가하였다. JNK1과 GSK-$3{\beta}$의 과발현은 각각 plasmid vector와 adenovirus vector를 이용하였다. 결 과 : PS-341 처치로 아포프토시스에 의한 세포생존율의 감소가 관찰되었다. PS-341 처치로 JNK가 활성화되었고, c-Jun의 발현이 유도되었다. Dominant negative JNK1의 과발현 또는 SP600125 전치치로 JNK의 활성화를 차단하면 PS-341에 의한 아포프토시스가 억제되었다. PS-341 처리로 JNK 활성화에 의존적으로 caspase 3의 활성화가 유도되었다. Caspase 활성화의 차단으로도 PS-341에 의한 아포프토시스가 억제되었다. PS-341에 의해 Akt가 활성화되었고, Akt 활성화의 차단으로 PS-341에 의한 아포프토시스가 심화되었다. PS-341에 의한 Akt 활성화로 GSK-$3{\beta}$가 불활성화되었다. Constitutively active GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 심화되었고, dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 감소되었다. Lithium chloride 전처치와 dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 JNK의 활성화와 c-Jun의 발현 증가가 억제되었다. 결 론 : 폐암세포주에서 PS-341에 의한 아포프토시스는 JNK/caspase 경로가 관여하며, 이는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화에 의해 억제되는 것으로 판단된다. 따라서 PS-341의 항암효과를 최대화하기 위해서는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화를 차단하는 치료법이 병행되어야 할 것으로 판단된다.

• 동의생리병리학회지
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• 제23권1호
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• pp.57-62
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• 2009

본 연구는 토복령의 항염증 및 세포보호 효과에 미치는 영향에 관한 것으로서 주요 내용은 다음과 같다. 본 실험에서는 세포독성, NO의 생성, PGE2, TNF-$\alpha$와 카탈라아제 농도, SOD, MAP kinase 등을 측정하였다. 본 실험의 결과 토복령 추출물은 세포 독성이 없었고 NO의 생성을 억제하며, 항염증과 세포보호 효과가 있었다. 그러나 이러한 효과에 대한 명확한 메커니즘에 대해서는 좀 더 연구가 필요하다는 내용이다.

• BMB Reports
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• 제50권12호
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• pp.634-639
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• 2017

We aimed to assess the anti-inflammatory and antioxidative properties of KHG26792, a novel azetidine derivative, in amyloid ${\beta}$ ($A{\beta}$)-treated primary microglial cells. KHG26792 attenuated the $A{\beta}-induced$ production of inflammatory mediators such as IL-6, $IL-1{\beta}$, $TNF-{\alpha}$, and nitric oxide. The levels of protein oxidation, lipid peroxidation, ROS, and NADHP oxidase enhanced by $A{\beta}$ were also downregulated by KHG26792 treatment. The effects of KHG26792 against the $A{\beta}-induced$ increases in inflammatory cytokine levels and oxidative stress were achieved by increasing the phosphorylation of $Akt/GSK-3{\beta}$ signaling and by decreasing the $A{\beta}-induced$ translocation of $NF-{\kappa}B$. Our results provide novel insights into the use of KHG26792 as a potential agent against $A{\beta}$ toxicity, including its role in the reduction of inflammation and oxidative stress. Nevertheless, further investigations of cellular signaling are required to clarify the in vivo effects of KHG26792 against $A{\beta}-induced$ toxicity.

• BMB Reports
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• 제51권7호
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• pp.362-367
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• 2018

A major feature of type 1 diabetes mellitus (T1DM) is hyperglycemia and dysfunction of pancreatic ${\beta}$-cells. In a previous study, we have shown that Tat-DJ-1 protein inhibits pancreatic RINm5F ${\beta}$-cell death caused by oxidative stress. In this study, we examined effects of Tat-DJ-1 protein on streptozotocin (STZ)-induced diabetic mice. Wild type (WT) Tat-DJ-1 protein transduced into pancreas where it markedly inhibited pancreatic ${\beta}$-cell destruction and regulated levels of serum parameters including insulin, alkaline phosphatase (ALP), and free fatty acid (FFA) secretion. In addition, transduced WT Tat-DJ-1 protein significantly inhibited the activation of $NF-{\kappa}B$ and MAPK (ERK and p38) expression as well as expression of COX-2 and iNOS in STZ exposed pancreas. In contrast, treatment with C106A mutant Tat-DJ-1 protein showed no protective effects. Collectively, our results indicate that WT Tat-DJ-1 protein can significantly ameliorate pancreatic tissues in STZ-induced diabetes in mice.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.217-223
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• 2010

To evaluate the antiallergic effect of fermented Ixeris sonchifolia (IS, family Compositae), we prepared IS kimchi, isolated lactic acid bacteria (LAB) from it, fermented IS with these LAB, and investigated their antiallergic effects. IS kimchi inhibited the passive cutaneous anaphylaxis (PCA) reaction induced by an IgE-antigen complex as well as the scratching behavior induced by compound 48/80 or histamine more potently than IS. When IS was fermented with LAB isolated from IS kimchi, its antiallergic effects was also increased. Of LAB used for fermentation, Lactobacillus brevis more potently increased the antiallergic effects. Its main constituents, chlorogenic acid and luteolin, potently inhibited the PCA reaction induced by the IgE-antigen complex as well as the pruritis induced by compound 48/80 or histamine. These constituents inhibited the expression of pro inflammatory and allergic cytokines, TNF-$\alpha$. and IL-4, and transcription factor NF-${\kappa}B$ activation induced by the IgE-antigen complex in RBL-2H3 cells, as well as the degranulation of RBL-2H3 cells induced by the IgE-antigen complex. Luteolin more potently inhibited these allergic reactions than chlorogenic acid. These findings suggest that the antiallergic effect of IS can be increased by LAB fermentation, and the fermented IS might improve allergic reactions such as pruritus, anaphylaxis, and inflammation.

• The Korean Journal of Physiology and Pharmacology
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• 제20권6호
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• pp.581-593
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• 2016

The advantages of monounsaturated fatty acids (MUFAs) on insulin resistance and type 2 diabetes mellitus (T2DM) have been well established. However, the molecular mechanisms of the anti-diabetic action of MUFAs remain unclear. This study examined the anti-hyperglycemic effect and explored the molecular mechanisms involved in the actions of fish oil- rich in MUFAs that had been acquired from hybrid catfish (Pangasius larnaudii${\times}$Pangasianodon hypophthalmus) among experimental type 2 diabetic rats. Diabetic rats that were fed with fish oil (500 and 1,000 mg/kg BW) for 12 weeks significantly reduced the fasting plasma glucose levels without increasing the plasma insulin levels. The diminishing levels of plasma lipids and the muscle triglyceride accumulation as well as the plasma leptin levels were identified in T2DM rats, which had been administrated with fish oil. Notably, the plasma adiponectin levels increased among these rats. The fish oil supplementation also improved glucose tolerance, insulin sensitivity and pancreatic histological changes. Moreover, the supplementation of fish oil improved insulin signaling ($p-Akt^{Ser473}$ and p-PKC-${\zeta}/{\lambda}^{Thr410/403}$), $p-AMPK^{Thr172}$ and membrane GLUT4 protein expressions, whereas the protein expressions of pro-inflammatory cytokines (TNF-${\alpha}$ and nuclear NF-${\kappa}B$) as well as p-PKC-${\theta}^{Thr538}$ were down regulated in the skeletal muscle. These data indicate that the effects of fish oil-rich in MUFAs in these T2DM rats were partly due to the attenuation of insulin resistance and an improvement in the adipokine imbalance. The mechanisms of the anti-hyperglycemic effect are involved in the improvement of insulin signaling, AMPK activation, GLUT4 translocation and suppression of pro-inflammatory cytokine protein expressions.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9915-9919
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• 2014

Lamellarin D (LamD) is a marine alkaloid with a pronounced cytotoxicity against a large panel of cancer cells, affecting cell growth and inducing apoptosis. However, the molecular mechanisms of action of this compound are poorly understood. In this study, the anticancer efficacy of LamD was investigated in human leukemia K562 cells. The results showed suppressed cell proliferation and induction of G0/G1-phase arrest,while expression of CDK1, and activity of smad3 and smad5 were reduced, but that of p27, p53 and STGC3 was increased. LamD induced cell apoptosis through activation of caspases-8/-3, inhibition of survivin and Bcl-2, suggesting that this compound may also act through a caspase-independent pathway. Moreover, LamD inhibited the secretion of TGF-${\beta}$, IL-$1{\beta}$, IL-6, IL-8 and other inflammatory cytokines and the transcriptional activity of transcription factor NF-${\kappa}B$ in human leukemia K562 cells.Taken together, our results suggest that LamD-mediated inhibition of leukemia cell proliferation may be related to the induction of apoptosis and the regulation of cell cycle, tumor-related gene expression and cytokine expression, which may provide a new way of thinking for the treatment leukemia.

• The Korean Journal of Physiology and Pharmacology
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• 제17권1호
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• pp.65-71
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• 2013

The transient receptor potential melastatin type 7 (TRPM7) channel is a widely expressed non-selective cation channel with fusion to the C-terminal alpha kinase domain and regarded as a key regulator of whole body $Mg^{2+}$ homeostasis in mammals. However, the roles of TRPM7 during osteoclastogenesis in RAW264.7 cells and bone marrow-derived monocyte/macrophage precursor cells (BMMs) are not clear. In the present study, we investigate the roles of TRPM7 in osteoclastogenesis using methods of small interfering RNA (siRNA), RT-PCR, patch-clamp, and calcium imaging. RANKL (receptor activator of NF-${\kappa}B$ ligand) stimulation did not affect the TRPM7 expression and TRPM7-mediated current was activated in HEK293, RAW264.7, and BMM cells by the regulation of $Mg^{2+}$. Knock-down of TRPM7 by siTRPM7 reduced intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increases by 0 mM $[Mg^{2+}]_e$ in HEK293 cells and inhibited the generation of RANKL-induced $Ca^{2+}$ oscillations in RAW264.7 cells. Finally, knock-down of TRPM7 suppressed RANKL-mediated osteoclastogenesis such as activation and translocation of NFATc1, formation of multinucleated cells, and the bone resorptive activity, sequentially. These results suggest that TRPM7 plays an essential role in the RANKL-induced $[Ca^{2+}]_i$ oscillations that triggers the late stages of osteoclastogenesis.

• The Korean Journal of Physiology and Pharmacology
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• 제15권6호
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• pp.397-403
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• 2011

The proliferation, migration, cytokine release, and contraction of airway smooth muscle cells are key events in the airway remodeling process that occur in lung disease such as asthma, chronic obstruction pulmonary disease, and cancer. These events can be modulated by a number of factors, including cigarette smoke extract (CSE). CSE-induced alterations in the viability, migration, and contractile abilities of normal human airway cells remain unclear. This study investigated the effect of CSE on cell viability, migration, tumor necrosis factor (TNF)-${\alpha}$ secretion, and contraction in normal human bronchial smooth muscle cells (HBSMCs). Treatment of HBSMCs with 10% CSE induced cell death, and the death was accompanied by the generation of reactive oxygen species (ROS). CSE-induced cell death was reduced by N-acetyl-l-cysteine (NAC), an ROS scavenger. In addition, CSE reduced the migration ability of HBSMCs by 75%. The combination of NAC with CSE blocked the CSE-induced reduction of cell migration. However, CSE had no effect on TNF-${\alpha}$ secretion and NF-${\kappa}B$ activation. CSE induced an increase in intracellular $Ca^{2+}$ concentration in 64% of HBSMCs. CSE reduced the contractile ability of HBSMCs, and the ability was enhanced by NAC treatment. These results demonstrate that CSE treatment induces cell death and reduces migration and contraction by increasing ROS generation in normal HBSMCs. These results suggest that CSE may induce airway change through cell death and reduction in migration and contraction of normal HBSMCs.

• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.315-320
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• 2013

Here, we show that radicicol, a fungal antibiotic, resulted in marked inhibition of inducible nitric oxide synthase (iNOS) transcription by the pancreatic beta cell line MIN6N8a in response to cytokine mixture (CM: TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$). Treatment of MIN6N8a cells with radicicol inhibited CM-stimulated activation of NF-${\kappa}B$/Rel, which plays a critical role in iNOS transcription, in a dose-related manner. Nitrite production in the presence of PD98059, a specific inhibitor of the extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) pathway, was dramatically diminished, suggesting that the ERK1/2 pathway is involved in CM-induced iNOS expression. In contrast, SB203580, a specific inhibitor of p38, had no effect on nitrite generation. Collectively, this series of experiments indicates that radicicol inhibits iNOS gene expression by blocking ERK1/2 signaling. Due to the critical role that NO release plays in mediating destruction of pancreatic beta cells, the inhibitory effects of radicicol on iNOS expression suggest that radicicol may represent a useful anti-diabetic activity.