• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.311-323
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• 2007

Receptor activation of nuclear factor ${\kappa}$ B ligand (RANKL)은 파골세포의 분화와 기능에 중요한 역할을 하는 단백질로 이들 물질의 조절에는 p38 MAP kinase가 관여한다. 그러나 치주인대 섬유모세포에서 RANKL 발현 시 p38 MAP kinase의 역할은 잘 알려져 있지 않다. 이에 이번 연구는 마우스 치주인대 섬유모세포의 $IL-1{\beta}-induced$ RANKL 발현과정에서 p38의 역할을 규명하고자 하여 다음과 같은 결과를 얻었다. 마우스 치주인대 섬유모세포에 $IL-1{\beta}$ (1ng/ml)의 자극은 수용성 RANKL의 합성을 증가시켰다. 수용성 RANKL의 합성은 p38 MAP kinase 억제제인 SB203580에 의해 농도 의존적으로 억제되었으나 다른 MAP kinase 억제제인 SP600125, JNK 억제제와 PD98059, ERK 억제제에 의해서는 수용성 RANKL의 합성이 조절되지 않았다. NF-kB 억제제에 의해서도 수용성 RANKL의 합성이 억제되지 않았다. RANKL 유전자의 발현은 $IL-1{\beta}$로 자극 시에는 대조군에 비해 약 5배의 발현 증가를 보였으나 SB203580으로 전처치 시 $IL-1{\beta}$ (1ng/ml)로 자극시보다 약 1.5배의 감소를 보였다. 그러나 SP600125, PD98059, 및 NF-kB 억제제로 전처치한 경우에는 $IL-1{\beta}$로 자극한 경우와 비슷한 수준을 보였다. $IL-1{\beta}$로 자극 시 RANKL 유전자의 반감기가 90분 이었으나 SB203580로 전처치 후 $IL-1{\beta}$로 자극 시 RANKL 유전자의 반감기는 60분으로 감소하였다. Cycloheximide 전처리 시 SB203580에 의한 RANKL 유전자 발현 억제가 관찰되지 않았다. 단백질 분석결과 p38 MAP kinase의 인산화 활성은 30분까지 증가하였으나 그 이후 감소하여 2시간째에는 그 발현이 미약하였다. SB203580로 전처치 후 $IL-1{\beta}$로 자극 시 p38 MAP kinase의 인산화 활성이 감소하였다. 이상의 결과는 p38 MAP kinase가 RANKL 유전자 조절에 중요한 역할을 담당하고 있음을 시사한다.

• Journal of Life Science
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• v.19 no.3
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• pp.299-304
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• 2009

Interleukin 27 (IL-27) was discovered as a heterodimeric cytokine of the IL-12 family, and is composed of two subunits - Epstein-Barr virus induced gene 3 (EBI3) and p28. It acts as a versatile cytokine in the early regulation of Th1 initiation and in the negative regulation of the Th2 factor GATA binding protein 3 (GATA-3). This cytokine is mediated by the IL-27 receptor (WSX-1), which is highly expressed on $CD4^+$ T lymphocytes and NK cells. We previously identified four polymorphisms in the human IL-27p28 gene and suggested that the polymorphism of IL-27p28 is associated with susceptibility to asthma. To determine whether these IL-27p28 SNPs are associated with susceptibility to allergic rhinitis, the genotype and allele frequencies of IL-27p28 SNPs were analyzed between allergic rhinitis patients and healthy controls. Although the genotype and allele frequencies of IL-27p28 SNPs in allergic rhinitis patients were not significantly different from those of the control group, there was a suggestive difference (P=0.037) between these groups in total serum IgE levels in the g.2905T>G SNP of the IL-27p28 gene. Our result implies that the g.2905T>G SNP of the IL-27p28 gene might have an affect on IgE production in allergic rhinitis patients.

• Tuberculosis and Respiratory Diseases
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• v.42 no.6
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• pp.862-870
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• 1995

Background: Oxygen free radicals have generally been considered as cytotoxic agents. On the other hand, recent results suggest that small nontoxic amounts of these radicals may act a role in intracellular signal transduction pathway and many efforts to reveal the role of these radicals as secondary messengers have been made. It is evident that the oxygen radicals are released by various cell types in response to extracellular stimuli including LPS, TNF, IL-1 and phorbol esters, all of which translocate the transcription factor $NF{\kappa}B$ from cytoplasm to nucleus by releasing an inhibitory protein subunit, $I{\kappa}B$. Activation of $NF{\kappa}B$ is mimicked by exposure to mild oxidant stress, and inhibited by agents that remove oxygen radicals. It means the cytoplasmic form of the inducible tanscription factor $NF{\kappa}B$ might provide a physiologically important target for oxygen radicals. At the same time, it is well known that LPS induces the release of oxygen radicals in neutrophil with the activation of $NF{\kappa}B$. From above facts, we can assume the expression of IL-8 and IL-$1{\beta}$ gene by LPS stimulation may occur through the activation of $NF{\kappa}B$, which is mediated through the release of $I{\kappa}B$ by increasing amounts of oxygen radicals. But definitive evidence is lacking about the role of oxygen free radicals in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. We conducted a study to determine whether oxygen radicals act a role in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. Method: Human peripheral blood monocytes were isolated from healthy volunteers. Time and dose relationship of $H_2O_2$-induced IL-8 and IL-$1{\beta}$ mRNA expression was observed by Northern blot analysis. To evaluate the role of oxygen radicals in the expression of IL-8 and IL-$1{\beta}$ mRNA by LPS stimulation, pretreatment of various antioxiants including PDTC, TMTU, NAC, ME, Desferrioxamine were done and Northern blot analysis for IL-8 and IL-$1{\beta}$ mRNA was performed. Results: In PBMC, dose and time dependent expression of IL-8 and IL-$1{\beta}$ mRNA by exogenous $H_2O_2$ was not observed. But various antioxidants suppressed the expression of LPS-induced IL-8 and IL-$1{\beta}$ mRNA expression of PBMC and the suppressive activity was most prominant when the pretreatment was done with TMTU. Conclusion: Oxygen free radical may have some role in the expression of IL-8 and IL-$1{\beta}$ mRNA of PBMC but that radical might not be $H_2O_2$.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.52-66
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• 2001

Background : NF-${\kappa}B$ is the most important transcriptional factor in IL-8 gene expression. Triptolide is a new compound that recently has been shown to inhibit NF-${\kappa}B$ activation. The purpose of this study is to investigate how triptolide inhibits NF-${\kappa}B$-dependent IL-8 gene transcription in lung epithelial cells and to pilot the potential for the clinical application of triptolide in inflammatory lung diseases. Methods : A549 cells were used and triptolide was provided from Pharmagenesis Company (Palo Alto, CA). In order to examine NF-${\kappa}B$-dependent IL-8 transcriptional activity, we established stable A549 IL-8-NF-${\kappa}B$-luc. cells and performed luciferase assays. IL-8 gene expression was measured by RT-PCR and ELISA. A Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation and an electromobility shift assay was done to analyze NF-${\kappa}B$ DNA binding. p65 specific transactivation was analyzed by a cotransfection study using a Gal4-p65 fusion protein expression system. To investigate the involvement of transcriptional coactivators, we perfomed a transfection study with CBP and SRC-1 expression vectors. Results : We observed that triptolide significantly suppresses NF-${\kappa}B$-dependent IL-8 transcriptional activity induced by IL-$1{\beta}$ and PMA. RT-PCR showed that triptolide represses both IL-$1{\beta}$ and PMA-induced IL-8 mRNA expression and ELISA confirmed this triptolide-mediated IL-8 suppression at the protein level. However, triptolide did not affect $I{\kappa}B{\alpha}$ degradation and NF-$_{\kappa}B$ DNA binding. In a p65-specific transactivation study, triptolide significantly suppressed Gal4-p65T Al and Gal4-p65T A2 activity suggesting that triptolide inhibits NF-${\kappa}B$ activation by inhibiting p65 transactivation. However, this triptolide-mediated inhibition of p65 transactivation was not rescued by the overexpression of CBP or SRC-1, thereby excluding the role of transcriptional coactivators. Conclusions : Triptolide is a new compound that inhibits NF-${\kappa}B$-dependent IL-8 transcriptional activation by inhibiting p65 transactivation, but not by an $I{\kappa}B{\alpha}$-dependent mechanism. This suggests that triptolide may have a therapeutic potential for inflammatory lung diseases.

• Journal of Acupuncture Research
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• v.21 no.2
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• pp.21-29
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• 2004

목적 : 본 연구는 Interleukin 4 Receptor (IL-4R) 유전자 다형성이 중풍의 발병과 관련이 있는지 알아보기 위해 수행되었다. 대상: 대구한의대학교부속 한방병원에 입원한 중풍환자 56명과 종합건진센터에 내원한 중풍 기왕력이 없는 건강인 83명을 대상으로 하였다. 방법 : 각 그룹에서 개개인마다 DNA를 분리 정제한 후 Taq polymerase로 증폭하여 한천 겔에서 전기영동을 하여 PCR 산물을 확인하였다. PCR 산물은 Pyrosequencing 과정을 통하여 IL4R의 유전형이 자동으로 판정되었다. 결과 : A/A, A/G, G/G의 세가지 유전자형이 검출되었으며 중풍군과 대조군 사이에 유의성 있는 차이가 발견되었다(p=0.005). 그러나 개별 allele 빈도에 있어서는 중풍군과 건강인 사이에 통계적인 유의성이 나타나지 않았다(p=0.995). 결론 : 이상의 결과를 통하여 IL4R 유전자 다형성은 중풍의 발병과는 관련성이 있는 것으로 사려되지만 더 많은 환자를 대상으로 다른 환경요인 또는 유전자와의 연관성에 대한 심도있는 연구가 필요하다고 하겠다.

• Korean Journal of Oriental Medicine
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• v.11 no.2
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• pp.113-140
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• 2005

1. 백렴 추출물은 부분적으로 종양세포인 HT1080, Hep G2, MCF-7 cell에 대해 세포독성을 보였으나, 섬유세포인 7250 cells과 종양세포인 CT-26 cells에서는 세포독성은 나타나지 않았다. 2. 백렴 추출물은 macrophage에서 생산되는 NO production을 부분적으로 증가시켰다. 3. 백렴 추출물은 macrophage 활성화와 관련 있는 IL-1과 iNOS의 유전자 발현을 증가시켰다. 4. 백렴 추출물은 NK cell의 cytotoxicity를 활성화시켰다. 5. 백렴 추출물은 NK cell 활성화와 관련있는 IL-1, IL-4, IL-10, IL-12, iNOS, IFN-${\gamma}$, TNF-${\alpha}$ 의 유전자 발현을 증가시켰다. 6. 백렴 추출물은 마우스의 비장세포에서 IL-10, IFN-${\gamma}$, TNF-${\alpha}$ 의 단백질 발현을 증가시켰다. 7. 백렴 추출물은 CT-26 cell에 의한 pulmonary colony를 대조군에 비해 유의성 있게 억제시켰다. 이상의 결과로 백렴 추출물은 macrophage와 NK cell의 활성화를 통해 면역 조절과 항암제로서의 유용성이 기대된다.

• Journal of Acupuncture Research
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• v.23 no.1
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• pp.39-51
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• 2006

목적 : 본 연구는 뇌경색에서 일반적으로 많이 사용하는 한방치료가 뇌경색 환자의 단일유전자 염기 다형성에 미치는 영향에 대하여 분석하였다. 2003년 3월부터 2003년 12월까지 경희대학교 한의과대학 부속한방병원 침구과에 입원한 뇌경색 환자 146명과 경희의료원 종합검진센터에 건강검진을 위해 내원한 건강인 192명을 대상으로 하였다. 방법 : 한국인 뇌경색 환자와 건강인에서 혈액을 채취하여 개인마다 DNA를 분리 정제하고 Taq polymerase로 증폭한 후 Pyrosequencing을 통하여 IL4R(interleukin 4 receptor)의 유전형을 관찰하였다. 결과 : 본 연구 결과 IL4R 유전자의 경우 한국인 뇌경색 환자군과 대조군 사이에 유의성 있는 차이가 나타나지 않았다. 결론 : 이상의 결과를 통하여 IL4R 유전자 다형성은 한국인에서 뇌경색의 발병에 관련이 적은 것으로 사려되며 더 많은 환자를 대상으로 다른 환경요인 또는 유전자와의 연관성에 대한 심도 깊은 연구가 필요할 것으로 사려된다.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.1
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• pp.144-160
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• 2007

목 적 : 이 연구는 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 사용되는 청폐화담탕(淸肺化痰湯)의 항염작용(抗炎作用)의 효과에 대해 알아보기 위해 시행되었다. 방 법 : 청폐화담탕(淸肺化痰湯)의 항염작용(抗炎作用)의 효과를 평가하기 위해 세포독성에 미치는 영향, NO, TNF-${\alpha}$, IL-l${\beta}$, IL-6 생성량에 미치는 영항, TNF-${\alpha}$, IL-l${\beta}$, IL-6 유전자 발현에 미치는 영향, iNOS, 염증cytokine 유전자 및 단백질 발현에 미치는 영향, PGE$_2$ 합성에 미치는 영향 및 NF-${\kappa}$B 활성에 미치는 영향에 대한 실험평가를 하였다. 결 과 : 청폐화담탕(淸肺化痰湯)은 MTT 분석을 통한 RAW 264.7 세포주의 생존력 평가에서 세포독성이 없었고, LPS로 유도된 RAW 264.7 세포주에서 NO 생성량을 농도 의존적으로 억제하였다. 청폐화담탕(淸肺化痰湯)은 400 g/ml 농도에서 LPS로 유도된 RAW 264.7 세포주에 대해 TNF-${\alpha}$, IL-1${\beta}$, IL-6 생성량을 각각 41.86${\pm}$2.26 %, 61.11${\pm}$2.54 %, 55.33${\pm}$3.65 % 억제하였으며 TNF-${\alpha}$, IL-1${\beta}$ 및 IL-6 유전자 발현을 농도 의존적으로 억제하였고, LPS로 유도된 RAW 264.7 세포주에서 iNOS와 COX-2 유전자 및 단백질 발현은 농도 의존적으로 억제하였다. 또한 그 농도에 따라 PGE$_2$ 생성량이 현저하게 억제하였고, LPS로 유도된 NF-${\kappa}$B 전사활성을 농도 의존적으로 억제함으로써 iNOS와 염증Cytokine 유전자 발현을 하향조절 하였다. 결 론 : 이상의 실험을 통해 청폐화담탕(淸肺化痰湯)은 LPS로 유도된 macrophage에서 NO와 염증Cytokine 생성량을 억제하였고 murine macrophage에서 NF-${\kappa}$B 활성을 억제함으로써 iNOS와 염증Cytokine 유전자 발현을 하향조절 하였다. 이러한 청폐화담탕(淸肺化痰湯)의 항염작용으로 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 응용할 수 있으리라 사료된다.

• Korean Journal of Biological Psychiatry
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• v.7 no.2
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• pp.147-151
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• 2000

Objective : Major depression is known to have immunologic dysfunctions, the recent studies revealed that cytokines including IL-6 and IL-$1{\beta}$ were increased in patients with major depression. Since molecular genetic methods have been progressed, this study was to investigate the relationship between major depression and immunologic aspects by analyzing polymorphism of IL-10 gene. Method : 92 patients with major depression were included and data of 146 normal controls obtained from the Catholic Hemopoietic Stem Cell Information Bank of Korea were used in this study. DNA was extracted from whole blood, thereafter amplified by polymerase chain reaction, and digested by Mae III After that procedure, we obtained and assessed RFLP of two alleles, IL-10T and IL-10C. All data were analyzed by ${\chi}^2$ test. Results : 1) There were no significant difference in genotype frequencies of $IL-10^*T/T$, $IL-10^*T/C$, and $IL-10^*C/C$ between major depression patients group and control group. 2) There were no significant difference in allelic frequencies of $IL-10^*T$ and $IL-10^*C$ between major depression patients group and control group. Conclusion : We did not verified the differences in frequencies of $IL-10^*T/^*IL-10^*C$ gene between the major depression patients group and control group, respectively. But the results of this study do not declare that the IL-10 gene has no association with major depression. We do suggest that further systematic studies including various clinical variables should be conducted.

• Journal of Life Science
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• v.30 no.2
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• pp.113-121
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• 2020

Macrophages are initiators for regulating a host's defenses to eliminate pathogens and trigger tissue repair. Macrophages are classified into two types: classically (M1) activated macrophages and alternatively (M2) activated macrophages. M1-phenotype macrophages directly or indirectly kill infectious organisms and tumor cells via pro-inflammatory responses, whereas M2-phenotype macrophages remodel wounded tissue through anti-inflammatory responses. In this paper, we investigated how Phellinus linteus hot water extract passed from Diaion HP-20 resin (PLEP) regulates polarization of M1-like or M2-like macrophages in human THP-1 cells. PLEP did not have cytotoxicity at a high concentration of 300 ㎍/ml. We observed morphological alteration of the THP-1 cells, which are stimulated by PLEP, LPS/INF-γ (M1 stimulators) or IL-4/IL13 (M2 stimulators). PLEP exposure induced morphology contiguous with LPS/INF-γ. qPCR was also performed to determine whether PLEP influences M1 or M2 polarization-related genes. M1-phenotype macrophage-specific genes, such as TNF-α, IL-1β, IL-6, IL-8, CXCL10 and CCR7, were enhanced by PLEP in a dose-dependent manner similar to LPS/INF-γ. Conversely, M2-phenotype-specific genes, such as MRC-1, DC-SIGN, CCL17 and CCL22, were suppressed by PLEP. PLEP also significantly up-regulated secretory inflammation cytokines related to M1 polarization of macrophages, including TNFα, IL-1β and IL-6, which was similar to the gene expression. Further, MAPK and NF-κB signaling were increased by treatment with PLEP, resulting in enhancement of cytokine secretion. PLEP might therefore be used as a promising booster of pro-inflammatory responses through M1 polarization of human THP-1 cells.