• The Korean Journal of Physiology and Pharmacology
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• 제27권5호
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• pp.471-479
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• 2023

Disulfiram (DSF), a medication for alcoholism, has recently been used as a repurposing drug owing to its anticancer effects. Despite the crucial role of dendritic cells (DCs) in immune homeostasis and cancer therapy, the effects of DSF on the survival and function of DCs have not yet been studied. Therefore, we treated bone marrow-derived DCs with DSF and lipopolysaccharide (LPS) and performed various analyses. DCs are resistant to DSF and less cytotoxic than bone marrow cells and spleen cells. The viability and metabolic activity of DCs hardly decreased after treatment with DSF in the absence or presence of LPS. DSF did not alter the expression of surface markers (MHC II, CD86, CD40, and CD54), antigen uptake capability, or the antigen-presenting ability of LPS-treated DCs. DSF decreased the production of interleukin (IL)-12/23 (p40), but not IL-6 or tumor necrosis factor-α, in LPS-treated DCs. We considered the granulocyte-macrophage colony-stimulating factor (GM-CSF) as a factor to make DCs resistant to DSF-induced cytotoxicity. The resistance of DCs to DSF decreased when GM-CSF was not given or its signaling was inhibited. Also, GM-CSF upregulated the expression of a transcription factor XBP-1 which is essential for DCs' survival. This study demonstrated for the first time that DSF did not alter the function of DCs, had low cytotoxicity, and induced differential cytokine production.

• Tuberculosis and Respiratory Diseases
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• 제60권6호
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• pp.663-672
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• 2006

연구배경: 미세먼지는 여러 가지 유기물과 무기물의 복합체로 그 구성 성분이 시간과 장소에 따라 다르고 모양과 크기도 일정하지 않으며, 특히 지름 $10{\mu}m$이하의 미세먼지 (particulate matter 10; $PM_{10}$)는 흡입이 가능한 입자의 크기여서 하부기관지 및 폐의 가스-교환부분까지 침착하여 호흡기계에 손상을 일으킬 수 있다. 본 연구에서는 황사에 포함된 $PM_{10}$과 비황사 시기에 포집된 $PM_{10}$이 폐상피세포주에 작용하여 전염증성 사이토카인(proinflammatory cytokine) 및 cytokine messenger RNA(mRNA)의 발현에 어떤 영향이 있는지를 관찰하여 기관지 천식과 만성 폐쇄성 폐질환등 호흡기 질환의 증상 악화기전에 미치는 역할을 규명하고자 하였다. 연구방법: 공기 포집기(HV 500F, sibata model)를 이용하여 황사와 비황사 기간에 하루 6시간씩 실외의 장소에서 대기분진을 membrane filter에 포집한 다음, $PM_{10}$입자를 추출하고 폐암 상피세포주인 A549 cells(한국세포은행주)에 $PM_{10}$을 농도에 맞게($10{\mu}g/ml$, $100{\mu}g/ml$, $500{\mu}g/ml$) 노출시켰다. 각각의 노출된 세포로부터 interleukin(IL)-$1{\alpha}$, $IL-1{\beta}$, IL-8, granulocyte macrophage colony stimulating factor(GM-CSF)의 mRNA를 역전사중합효소연쇄반응(reverse transcriptase polymerase chain reaction; RT-PCR) 방법으로 측정하였다. 결 과: 황사 및 비황사 기간 중 포집된 $PM_{10}$을 가했을 시 가하지 않은 대조군에 비하여 $IL-1{\alpha}$, $IL-1{\beta}$, IL-8, granulocyte macrophage colony stimulating factor (GM-CSF)의 m'RNA와 cytokine의 발현이 유의하게 높았으며, 황사 기간의 고농도의 $PM_{10}$에 노출된 세포의 $IL-1{\alpha}$ mRNA는 비황사 기간의 $PM_{10}$에 노출된 세포의 mRNA보다 증가되어 있었다. 결 론: $PM_{10}$은 A549 cells에서 전염증성 사이토카인의 발현을 증가시키고 비황사 기간보다 황사 기간 중 대기 중에서 채취한 $PM_{10}$에 노출된 A549 cells에서 일부의 전염증성 사이토카인의 mRNA발현을 더욱 증가시키는 것을 알 수 있었다. 따라서 황사 기간의 $PM_{10}$에 의한 일부의 전염증성 사이토카인의 발현 증가가 만성 호흡기 질환의 증상 악화기전에 연관되어 있을 가능성을 시사하였다.

• 대한한의학회지
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• 제33권4호
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• pp.42-49
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• 2012

Objectives: The purpose of this study was to investigate effects of Red Ginseng-Ejung-tang Water Extract (ER) on cytokine production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods: Levels of various cytokines such as interleukin (IL)-6, IL-10, IL-2, IL-12p70, vascular endothelial growth factor (VEGF), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-2, keratinocyte-derived chemokine (KC), tumor necrosis factor (TNF)-alpha, granulocyte macrophage colony-stimulating factor (GM-CSF) were measured by high-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. Results: ER significantly decreased levels of IL-6, IL-10, IL-2, IL-12p70, VEGF, and MCP-1 for 24 hrs incubation at the concentrations of 25, 50, and $100{\mu}g/mL$ in LPS-induced RAW 264.7 cells (P < 0.05). But ER did not exert significant effects on production of MIP-2, KC, TNF-${\alpha}$, and GM-CSF in LPS-induced RAW 264.7 cells. Conclusions: These results suggest that ER has an anti-inflammatory property related with its inhibition of cytokine production in LPS-induced macrophages.

• Toxicological Research
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• 제31권4호
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• pp.371-392
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• 2015

TS-DP2 is a recombinant human granulocyte colony stimulating factor (rhG-CSF) manufactured by TS Corporation. We conducted a four-week study of TS-DP2 (test article) in repeated intravenous doses in male and female Sprague-Dawley (SD) rats. Lenograstim was used as a reference article and was administered intravenously at a dose of $1000{\mu}g/kg/day$. Rats received TS-DP2 intravenously at doses of 250, 500, and $1000{\mu}g/kg/day$ once daily for 4 weeks, and evaluated following a 2-week recovery period. Edema in the hind limbs and loss of mean body weight and body weight gain were observed in both the highest dose group of TS-DP2 and the lenograstim group in male rats. Fibro-osseous lesions were observed in the lenograstim group in both sexes, and at all groups of TS-DP2 in males, and at doses of TS-DP2 $500{\mu}g/kg/day$ and higher in females. The lesion was considered a toxicological change. Therefore, bone is the primary toxicological target of TS-DP2. The lowest observed adverse effect level (LOAEL) in males was $250{\mu}g/kg/day$, and no observed adverse effect level (NOAEL) in females was $250{\mu}g/kg/day$ in this study. In the toxicokinetic study, the serum concentrations of G-CSF were maintained until 8 hr after administration. The systemic exposures ($AUC_{0-24h}$ and $C_0$) were not markedly different between male and female rats, between the administration periods, or between TS-DP2 and lenograstim. In conclusion, TS-DP2 shows toxicological similarity to lenograstim over 4-weeks of repeated doses in rats.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2523-2526
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• 2012

Introduction: Febrile neutropenia is a relatively frequent event in cancer patients treated with chemotherapy and improvement in absolute neutrophil count (ANC) has been linked directly to improved outcome. Evaluation of granulocyte colony stimulating factors (GCSFs) for treatment has shown reduced incidences of episodes of prolonged neutropenia and protracted hospitalization. To determine absolute neutrophil counts with GCSF in febrile neutropenic cancer patients admitted to a tertiary care centre and to co-relate the improvement in ANC with mortality and hospital discharge. Methods: A prospective cross sectional study was carried at an oncology ward at Aga Khan University hospital from January 2010 to June 2011. All adult patients who were admitted and treated with GCSF for chemotherapy induced febrile neutropenia were included. Multivariable regression was conducted to identify the factors related with poor outcomes. Results: A total of 131 patients with febrile neutropenia were identified with mean age of 43.2 (18-85) years, 79 (60%) being ${\leq}50$. Seventy-five (57%) had solid tumors and 56 (43%) hematological malignancies, including lymphoma. Fifty seven (43.5%) had an ANC less 100 cells/$mm^3$, 34 (26%) one between 100-300 cells/$mm^3$ and 40 (31%) an ANC greater than 300 cells/$mm^3$. Thirty (23%) patients showed ANC recovery in 1-3 days, and 74(56%) within 4-7 days. Thirteen (10%) patients showed no recovery. The overall mortality was 18 (13.7%) patients. The mean time for ANC recovery seen in hematological malignancies was 6.34 days whereas for solid tumors it was 4.88 days. Patients with ANC <100 cells/$mm^3$ were more likely to die than patients with ANC >300 cells/$mm^3$ by a factor of 4.3. Similarly patients >50 years of age were 2.7 times more likely to die than younger patients. Conclusion: Our study demonstrated that use of GCSF, in addition to intravenous antibiotics, in treatment of patients with chemotherapy induced febrile neutropenia accelerates neutrophil recovery, and shortens antibiotic therapy and hospitalization. We propose to risk classify the patients at the time of admission to evaluate the cost effectiveness of this approach in a resource constrained setup.

• BMB Reports
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• 제45권12호
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• pp.742-747
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• 2012

Granulocyte colony-stimulating factor (G-CSF) is used for heart failure therapy and promotes myocardial regeneration by inducing mobilization of bone marrow stem cells to the injured heart after myocardial infarction; however, this treatment has one weakness in that its biological effect is transient. In our previous report, we generated 5 mutants harboring N-linked glycosylation to improve its antiapoptotic activities. Among them, one mutant (Phe140Asn) had higher cell viability than wild-type hG-CSF in rat cardiomyocytes, even after treatment with an apoptotic agent ($H_2O_2$). Cells treated with this mutant significantly upregulated the antiapoptotic proteins, and experienced reductions in caspase 3 activity and PARP cleavage. Moreover, the total number of apoptotic cells was dramatically lower in cultures treated with mutant hG-CSF. Taken together, these results suggest that the addition of an N-linked glycosylation was successful in improving the antiapoptotic activity of hG-CSF, and that this mutated product will be a feasible therapy for patients who have experienced heart failure.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.89-92
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• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.

• 동의생리병리학회지
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• 제18권6호
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• pp.1896-1903
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• 2004

Anglicae dahuricae Radix (ADR), the dried roots of Angelica Dahurica Benth et Hook (Umbelliferae), is a traditional herbal medicine used to treat colds, headache, skin diseases such as acne and possess analgesic, antipyretic and drainage effects. In the present study, the author evaluated the effect of ADR on regulation of inflammatory reaction. ADR reduced the ear-swelling responses derived from compound 48/80 in dose-dependent manner significantly. ADR inhibited the PMA plus A23187-induced productions of IL (Interleukin)-8, IL-1β, granulocyte macrophage-colony stimulating factor (GM-CSF) and tumor necrosis factor (TNF)-α from human mast cells (HMC)-1. In addition, ADR blocked PMA plus A23187-induced ERK1/2 phosphorylation. Ⅰ suggest that ADR regulates inflammatory reaction through inhibition of inflammatory cytokines such as IL-8, IL-1β and GM-CSF.

• Advances in Traditional Medicine
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• 제5권1호
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• pp.69-74
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• 2005

'SecSec' has been used for the purpose of prevention and treatment of throat diseases such as sore throat, cough, bronchial asthma and allergic asthma in Korea. However, its effect in experimental models remains unknown. To investigate the biological effect of SecSec, we examined cytotoxicity and secretion of inflammatory cytokines on human leukemic mast cell line, HMC-1, stimulated with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. SecSec by itself had no cytotoxicity on HMC-1. When SecSec (1 mg/ml) was added, the secretion of tumor necrosis factor-alpha $(TNF-{\alpha})$, interleukin (IL)-6, and granulocyte macrophage-colony stimulating factor (GM-CSF) was significantly inhibited about 47.20%, 25.55%, and 46.43%, respectively on PMA plus A23187-stimulated HMC-1 cells. But SecSec did not inhibit IL-8 secretion. These findings may help understanding the mechanism of action of this medicine leading to control activated mast cells on allergic inflammatory condition like asthma.

• 대한약침학회지
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• 제15권1호
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• pp.7-11
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• 2012

Aim: Historically, mineral compound herbal medicines have long been used in treatments of immune-related diseases in Korea, China and other Asian countries. In this study, we investigated the anti-inflammatory effect of egg white combined with chalcanthite (IS4) on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: RAW 264.7 cells cultured with LPS and various concentrations of IS4 were analyzed to determine the production of pro-inflammatory cytokines and mediators by using enzyme-linked immune sorbent assays (ELISAs). Results: IS4 concentration inhibited the production of interleukin-1beta (IL-$1{\beta}$), interleukin-6 (IL-6), and granulocyte-macrophage colony-stimulating factor (GM-CSF) induced by LPS. IS4 at high concentrations (25 and 50 ${\mu}g/ml$) inhibited, in concentration-dependent manner, the expression of tumor necrosis factor-alpha (TNF-${\alpha}$) stimulated by LPS. Conclusion: IS4 has shown an anti-inflammatory effect in RAW 264.7 cells.