기니피그에서 정원창막에 투여된 IL-1β에 의한 청력변화와 그 투과에 관한 연구

이정구,장용주,조정석,황의진 대한이비인후과학회 1999 대한이비인후과학회지 두경부외과학 Vol.42 No.6

Effects of TGF-β, TNF-α, IL-β and IL-6 alone or in combination, and tyrosine kinase inhibitor on cyclooxygenase expression, prostaglandin E₂production and bone resorption in mouse calvarial bone cells

Park, Young-Guk,Kang, Sung-Koo,Kim, Won-Jin,Lee, Youn-Choon,Kim, Cheorl-Ho KYUNG HEE UNIVERSITY MEDICAL CENTER 2005 고황의학상 수상논문집 Vol.21-22 No.-

Cyclooxygenase-2 (COX-2) and tyrosine kinase, which are involved in the biosynthesis of prostaglandin E₂ (PGE₂) in mouse calvarial osteoblasts, are stimulated by cytokine interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and/or interleukin-6 (IL-6). IL-1β and IL-6 and, to a lesser extent, TNF-α, enhances COX-2 mRNA levels in calvarial osteoblasts. Simultaneous treatment with IL-6 and IL-1β and TNF-α resulted in enhanced COX-2 mRNA levels accompanied by the cooperative stimulation of PGE₂ biosynthesis compared to cells treated with IL-1β or TNF-α or IL-6 alone. In contrast, the presence of TGF-β reduced COX-2 mRNA level, PGE₂ biosynthesis and bone resorption induced by IL-1β, TNF-α, IL-6 or a combination thereof. However, neither IL-1β, TNF-α, IL-6 nor a combination of IL-1β, TNF-α, IL-6 enhanced COX-1 mRNA levels in calvarial osteoblasts. A novel Src tyrosine kinase inhibitor, Herbimycin A (HERB), reduced COX-2 mRNA levels as well as PGE₂ production induced by IL-1β, TNF-α and IL-6 or a combination of IL-1β, TNF-α, IL-6, whereas COX-1 mRNA levels remained unaffected. Finally, HERB was found to inhibit in vitro bone resorption. These results indicate that the cooperative effects of IL-β, TNF-α, IL-6 on PGE₂ production are due to the enhanced expression of the COX-2 gene and that tyrosine kinase(s) are involved in COX-2 signal transduction in mouse calvarial osteoblasts. Thus, the Src family of kinase inhibitors may be useful in treating diseases associated with elevated bone loss.

간 절제후의 IL-1β , IL-6 및 TNF-α의 변화

김홍진,윤성수,전용성,심민철,권굉보 大韓消化器病學會 1997 大韓消化器病學會誌 Vol.29 No.2

Effect of IL-6 on Osteoclast Generation and Regulation of Its Production in Osteoblastic Cells

Shin, Dong-In,Baek, Jeong-Hwa,Ko, Seung-Hee,Kim, Gwan-Shik The Official Publication of Korean Academy of Oral 1996 International Journal of Oral Biology Vol.20 No.1

Interleukin-6 (IL-6) is a multifunctional cytokine that known to be synthesized by osteoblasts and suggested to be an important regulator of bone resorption in some physiologic and/or pathologic conditions. However, the mechanism of cation of IL-6 responsible for bone resorption in vitro and role as a possible mediator of bone resorptive action of systemic or local osteotropic agents are not clear yet. Therefore, to further understand the mechanism of IL-6 action on bone resorption, we examined the effects of IL-6 on the generation of osteoclast-like cells using mouse bone marrow culture system. We also observed the regulation of IL-6 secretion in human osteoblastic cells by several cytokines and systemic osteotropic hormones, known to stimulate or inhibit bone resorption. To observe the effects of IL-6 on the generation of osteoclast-like cells, mouse bone marrow cells were isolated from femurs and tibiae of 4 to 6 week-old mice, plated at 3.0×10^6 cells/well in 24-well plates and cultured for 8 days. In order to examine the role of IL-6 in osteoclast-like multinucleated cell (MNC) generation, bone marrow cells were treated with IL-6 alone or in combination with prostaglandin E_2(PGE_2). After culture, cells were stained for tartrate-resistant acid phosphatase (TRAP), a marker enzyme of osteoclasts, according to the modified method of Burstone. The TRAP-positive MNCs, which have 3 or more nuclei, were counted. Also we observed the regulation of IL-6 secretion in human osteoblastic cells (MG-63) by several osteotropic agents using the quantitative enzyme-linked immunosorbent assay. The present study showed that IL-6 alone did not generate TRAP-positive MNCs, but significantly enhanced the generation of TRAP-positive MNCs induced by PGE_2(10^-6 M) in a dose-dependent manner. Though the augmenting effect of IL-6 was significant through all the culture period, it was greater at later period of culture. And augmenting effect of IL-6 on generation of TRAP-positive MNCs significantly appeared from the 6th-day of culture. These results suggest that IL-6 enhances the effect of PGE_2 on TRAP-positive MNC generation by stimulating both differentiation and fusion of osteoclast precursors. On the otherhand, bone resorbing cytokines such as interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α) stimulated the secretion of IL-6 from MG-63 cells, but interferon-γ(IFN-γ), known to be an inhibitor of bone resorption, significantly inhibited the TNF-α-stimulated secretion of IL-6. However, parathyroid hormone, 1,25(OH)_2-vitamin D_3, and PGE_2, well known bone resorbing agents, did not induce the secretion of IL-6 from MG-63 cells. Taken together, these results suggest that IL-6 may play an important role in bone resorption by regulating the generation of osteoclasts and at least partly, mediate the bone modulating effects of several cytokines.

신생 백서의 저산소성 허혈성 뇌손상에서 Pentoxifylline이 IL-1β 와 TNF-α mRNA 유전자 발현에 미치는 영향

김경범(Kyung Burm Kim),전지현(Gi Hyun Jeon),김영래(Young Rae Kim),이정화(Jung Hwa Lee),이기형(Kee Hyoung Lee),은백린(Baik Lin Eun),김순겸(Soon Kyum Kim) 대한소아신경학회 1999 대한소아신경학회지 Vol.7 No.2

목 적 : Interleukin-1β(IL-1β)와 Tumor necrosis factor-α(TNF-α)는 다양한 기능을 가진 사이토카인이며, 급성 혹은 만성 뇌손상의 매개체로의 역할이 점차 확인되고 있으나 이들의 정확한 역할에 대해서는 아직 논란이 많다. 최근 연구에서는 Pentoxifylline(PTX)을 이용하여 미성숙 실험 동물 모델에서 저산소상 허혈성 뇌손상을 완화시킬수 있었다. PTX 과 관련된 실험적 연구는 많으나 미성숙 뇌에서 뇌보호 작용기전에 관련된 보고는 매우 드물다. 이에 저자들은 신생 백서의 저산소성 허혈증에서 PTX에 의한 대뇌보호 작용 기전 중 IL-1β와 TNF-α 의 유전자 발현에 미치는 영향을 알아보기 위하여 본 연구를 시행하였다. 방 법 : 7일괸 Sprague-Dawley 백서를 사용하여 Isoflurane 마취하에 우측 총경동맥을 관찰하고 약 1시간 동안 회복시킨 후 36℃ 보육기 안에 있는 유리용기에서 3시간 동안 8% 산소를 노출시켰다. 실험군은 저산소 노출 직전과 직후 PTX를 각각 40mg/kg를 복강내 투여하였고, 대조군에게는 약물대신 Phosphate buffer solution(PBS)을 사용하였다. 저산소 및 허혈 후 4시간째 동물을 희생시키고 대뇌를 적출한 뒤 좌측과 우측 대뇌를 각각 분리하여 total RNA를 추출하였다. 랜덤 헥사머를 이용한 역전사 과정을 통해 얻어진 cDNA를 사용하여 PCR을 통해 IL-1β와 TNF-α를 증폭하고 동시에 GAPDH를 증폭하여 표준화하였다. 결 과 : 대조군에서 IL-1β와 TNF-α mRNA는 좌우측 모두 발현되었으나 우측 반구에서 현저하게 나타났고, PTX 투여군의 좌측 반구에서 발현되지 않았고 우측 반구에서도 현저히 감소하였다. 결 론 : PTX 투여로 신생 백서의 저산소성 허혈증에 의해 유발된 IL-1β와 TNF-α의 유전자 발현이 감소되었다. 이러한 결과는 PTX의 여러 약리 작용중 IL-1β와 TNF-α 유전자 발현을 특이하게 억제하고 미성숙 실험 동물 모델에서 저산소성 허혈증 뇌손상의 방어 효과와 관련이 있으리라 생각되며 또한 IL-1β와 TNF-α가 신생아의 저산소성 허혈증 뇌병증에서 중요한 매개체라는 사실을 시사한다. Purpose : Interleukin-1β(IL-1β) and Tumor necrosis factor-α(TNF-α) are multifunctional cytokines that may play important roles both in the normal development of central nervous system and in the response of brain to diverse forms of injury. IL-1β and TNF-α have potent proinflammatory action and the potential to modulate cell growth. Cerebral hypoxia-ischemia selectively stimulates IL-1β and TNF-α gene expression in brain regions susceptible to irreversible injury in perinatal rats. Pentoxifylline, a cAMP phosphodiesterase inhibitor, attenuates hypoxic-ischemic brain injury in immature rats and inhibits TNF-α expression at the transcription level. We hypothesize that pentoxifylline would attenuate the expression of IL-1β and TNF-α mRNA gene expression on hypoxic-ischemic brain injury in immature rats. Methods : To elicit focal hypoxic-ischemic brain injury, 7-d-old(P7) rats underwent right carotid artery ligation, followed by 3 hr of hypoxia(fractional concentration of inspired O₂=0.08). In 3 rats, pentoxifylline(40mg/kg) was injected into the intraperitoneal cavity immediately before and after hypoxia. The other 4 rats were given PBS solutions. IL-1β and TNF-α mRNA content were measured by reverse transcription followed by polymerase chain reaction amplification(RT-PCR) in the samples prepared from the lesioned and contralateral hemispheres killed 4 hr post-hypoxia. cDNA were amplified with primers specific for IL-1beta and TNF-alpha. and also amplified with GAPDH primers which served as an internal control. Results : In control group, hypoxia-ischemia induced IL-1β and TNF-α mRNA expression from the lesioned hemisphere in immature rat brain. In pentoxifylline treated group, IL-1β and TNF-α mRNA expression were attenuated at 4 hr post hypoxia- ischemia. Conclusion : Preteatment with pentoxifylline decreased incidence and severity of hypoxic-ischemic injury in immature rat brain. Pentoxifylline attenuated the expression of IL-1β and TNF-α gene on hypoxic-ischemic injury in immature rat brain. IL-1β and TNF-α may play important roles in the response of the developing brain to acute hypoxic-ischemic injury.

IL-β의 인슐린 분비 자극효과와 그 과정에 관여하는 인자들

정인경,오승훈,강동묵,정재훈,민용기,이명식,이문규,김광원 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.24 No.4

연구배경: IL­1β는 용량과 노출시간에 따라 인슐린분비에 대해 상반되는 효과를 가지고 있다. IL­1β의 인슐린 분비 억제효과는 제1혁명 당뇨병의 자가면역 기전과 관련되어 잘 알려져 있으나 인슐린 자극효과에 대해서는 아직 명백히 밝혀지지 않았다. 이에 저자 등은 IL­1β의 다양한 농도에 따라 백서의 췌도세포에서 인슐린 분비에 미치는 영향을 살펴보고, 그 기전으로 인슐린 생합성, iNOS의 발현, 칼슘통로의 활성도 변화여부를 알아보고자 하였다. 방법:200∼300g인 수컷 Sprague­Dawley 백서의 췌도를 변형된 Lacy&Kostianovsky's 방법으로 분리한 후 IL­1β의 다양한 농도(0, 0.5, 5, 50, 500pmol/L)에 2, 6, 24시간 노출시켜 췌도세포의 형태, 생존능을 관찰하고 인슐린 분비능 및 췌도세포내 인슐린 함량을 측정하였으며, 전전구 인슐린 mRNA발현, iNOS mRNA발현을 RT­PCR을 통해 확인하였고, 세포의 칼슘 통로 활성도 변화 여부를 측정하였다. 결과:1) IL­1β에 노출되 췌도의 생존능:2시간 노출시는 대조군과 차이가 없었으나, 6시간 고농도군과 24시간 모든 군에서 생존능이 감소되었다. 2)인슐린 분비능은 IL­1β를 2시간 5poml/L이상의 고농도와 6시간 0.5pmol/L 저농도 처리시 대조군에 비해 의미있게 증가하였으나, 6시간과 24시간 5poml/L이상의 고농도에서는 의미있게 인슐린의 분비가 억제 되었다. 3)췌도내 인슐린양의 변화는 IL­1β의 시간과 농도에 따라 배지내의 인슐린 변화와 비슷한 경향을 보였으나 통계학적으로 의미있는 차이는 없었다. 4)전전구인슐린의 mRNA발현은 2시간 50pmol/L이상 고농도의 IL­1β에서 의미있게 증가하였고, 6시간 처리군에서는 의미있는 차이는 없었으나, 24시간에서 IL­1β의 용량의존적으로 억제되었다. 5)iNOS mRNA는 IL­1β 처리 후 2시간부터 발현되기 시작하여, 6시간에 최고에 달한 후 24시간에는 점차 감소하였다. IL­1β의 처리시간과 무관하게 5poml/L이상의 고농도에서 용량에 따라 발현이 증가하였다. 6)칼슘통로 활성도는 IL­1β의 농도나 시간에 따라 유의한 차이가 없었다. 결론: IL­1β는 단시간 고용량이나 장기간 저용량에서 인슐린 분비와 생합성을 증가시키며, 이런 효과는 iNOS나 칼슘 통로 활성도 변화와는 무관한 것으로 생각된다. Background : The inhibitory effort of IL-1β on the insulin secretion has been validated in pathogenesis of type 1 diabetes, but complex results about the stimulatory effect of IL-1β have been reported. The aims of this study are to clarify the effects of IL-1βon insulin secretion of pancreatic islets and to investigate the mechanisms in terms of preproinsulin synthesis, inducible NOS expression, and calcium channel activity. Method : Islets were isolated from male Sprague-Dawley (SD) rat by modified Lacy-Kostianovsky's method. After islets were treated with different concentrations (0, 0.5, 5, 50, 500 pmol/L) and exposure time (2, 6, 24 hours) of IL-1β, morphology, viability, static stimulation of insulin to glucose, insulin content, preproinsulin mRNA expression, iNOS mRNA expression and calcium channel activity were measured. Results : 1) Viability o islets was reduced in high concentrations of long term exposure of IL-1β. 2) Insulin secretion was stimulated in islets treated with 5, 50, and 500 pmol/L of IL-1β for 2 hours. 3) Insulin content was not significantly different regardless of concentration and exposure time of IL-1β. 4) Preproinsulin mRNA expression increased in islets treated with 50, 500 pmol/L of IL-1β for 2 hours. After 24 hours, it decreased in dose dependent manner. 5) iNOS mRNA expression was detectable after 2 hours in the presence of IL-1β, peaks at 6 hour and decreased after 24 hours. It was increased above 5 pmol/L of IL-1β in dose dependent manner. 6) Activities of the voltage-dependent Ca^2+ channels were not different among groups. Conclusion : IL-1β plays a positive role in terms of insulin secretion and insulin synthesis in high concentration of short term or low concentration of long term. These effects of IL-1β might be neither dependent of iNOS pathway nor Ca^2+ channel activity (J Kor Diabetes Asso 431~443, 2000).

DPIE [2-(1,2-diphenyl-1H-indol-3-yl)ethanamine] Augments Pro-Inflammatory Cytokine Production in IL-1β-Stimulated Primary Human Oral Cells

Ahn, Sun-Hee,Lee, Jin-Kyung,Kim, Nam Doo,Kim, Seok-Ho,Lee, Sunwoo,Jung, Seunggon,Chay, Kee-Oh,Lee, Tae-Hoon MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.7

<P>Interleukin-1β (IL-1β) is a prominent pro-inflammatory cytokine that is implicated in a variety of autoimmune diseases and plays an important role in host defense against infections. IL-1β activity increases with its increasing binding capacity to IL-1 receptors (IL-1Rs). Thus, numerous studies have targeted the discovery of molecules modulating the interactions between IL-1β and IL-1R1. We have conducted an IL-1R1 structure-based virtual screening to identify small molecules that could alter IL-1β activity, using in silico computational analysis. Sixty compounds from commercial libraries were predicted to bind to IL-1R1, and their influence on cytokine production in IL-1β-stimulated gingival fibroblasts (GFs) was determined. Of these, only (2-(1,2-diphenyl-1H-indol-3-yl)ethanamine (DPIE) showed a synergistic increase in inflammatory molecules and cytokine production (IL-6, IL-8, and COX-2) at both mRNA and protein levels in IL-1β-stimulated GFs. The enhancing activity of DPIE in IL-1β-induced cytokine production increased in a dose-dependent manner without cytotoxicity. This pattern was also observed in IL-1β-stimulated primary human periodontal ligament cells (PDLs). Furthermore, we measured the impact of DPIE on the IL-1β–IL-1R1 system using surface plasmon resonance and demonstrated that DPIE increased the binding affinity of IL-1β to IL-1R1. These data indicate that DPIE boosts IL-1β signaling by enhancing the binding of IL-1β to IL-1R1 in oral primary cells.</P>

Interleukin-1B(1L-1B) polymorphisms and gastric mucosal levels of IL-Iβ cytokine in Korean patients with gastric cancer

Chang, Young-Woon,Jang, Jae-Young,Kim, Nam-Hoon,Lee, Jae Won,Lee, Hyo Jung,Jung, Woon Won,Dong, Seok-Ho,Kim, Hyo-Jong,Kim, Byung-Ho,Lee, Joung-Il,Rin Chang KYUNG HEE UNIVERSITY MEDICAL CENTER 2006 고황의학상 수상논문집 Vol.21-22 No.-

Interleukin-1B and IL-1 receptor antagonist gene polymorphisms are associated with an increased risk of gastric cancer (GC) in Caucasian populations. However, recent studies could not find any association between IL-1B-511T polymorphism and the risk of GC in Asians. We tested for an association between IL-1 loci polymorphisms with increased gastric mucosal levels of IL-1β and an increased risk of developing GC in a Korean population. Polymorphisms of IL-1A-889, IL-1B-31, IL-1B-511 and IL-1RN were genotyped in 434 controls and 234 patients with GC. Mucosal IL-1β cytokine was measured using an ELISA. The frequencies of IL-1A, IL-1B-511, IL-1B-31 and IL-1RN were not statistically different between controls and all patients with GC. After subclassification of GC, only patients with intestinal-type GC showed a higher frequency of IL-1B-31T homozygotes (OR = 2.2; 95% CI = 1.1-4.3) compared with controls. Risk was also significantly increased in these patients for IL-1B-31T homozygotes compared with patients with diffuse-type GC (OR = 3.4; 95% CI = 1.5-7.7). As in Caucasian populations, linkage disequilibrium between IL-1B-31 and IL-1B-511 was nearly complete, but the pattern of haplotype related to the risk of GC (IL-1B-31T/IL-1B-511C) was opposite (IL-1B-511T/IL-1B-31C). Mucosal IL-1β levels in H. pylori-infected GC patients were higher in patients homozygous for IL-1B-31T compared with IL-1B-31C/T and IL-1B-31C/C. Thus, the combined effects of H. pylori infection and IL-1B-31T/IL-1B-511C polymorphisms with enhanced mucosal IL-1β production contributed to the development of intestinal-type GC in this Korean population.

마우스 골수세포 배양시 수종 국소조절인자가 파골-유사세포 형성에 미치는 영향

채한정,채수완,김형룡 한국노화학회 1998 한국노화학회지 Vol.8 No.3

SW1353 연골세포에서 기능 성분이 증대된 보스웰리아 검레진 추출물의 항골관절염 효과 연구

정재인(Jae In Jung),이현숙(Hyun Sook Lee),김룡(Ryong Kim),김은지(Eun Ji Kim) 한국식품영양과학회 2023 한국식품영양과학회지 Vol.52 No.5

본 연구는 인체유래 연골세포인 SW1353 세포에 IL-1β를 처리하여 유도한 골관절염 세포 모델에서 인도산 보스웰리아 검레진을 주정으로 추출 후 지방 제거 공정을 추가하여 제조한 보스웰리아 검레진 추출물인 FJH-UBS의 항골관절염 효능을 평가하기 위해 실시하였다. SW1353 세포에 IL-1β 처리 시 염증매개물질인 NO/iNOS, PGE2/COX-2, IL-6 및 TNF-α 발현과 연골 기질 분해에 기여하는 MMPs(MMP-1, MMP-3, MMP-13) 발현이 증가하였다. 또한 IL-1β 처리에 의해 연골의 구성성분인 aggrecan과 type Ⅱ collagen 발현이 감소하였다. FJH-UBS는 IL-1β에 의해 증가한 NO/iNOS, PGE2/COX-2, IL-6, TNF-α, MMP-1, MMP-3 및 MMP-13 발현을 감소시켰고, IL-1β에 의해 감소한 aggrecan과 type Ⅱ collagen 발현을 증가시켰다. 골관절염의 염증반응과 연골 퇴화를 조절하는 주요한 신호전달체계인 MAPK(ERK, p38, JNK)와 NF-κB(p65, Iκ-Bα)의 인산화는 IL-1β에 의해 증가하였고, 이는 FJH-UBS에 의해 감소하였다. 이 결과는 FJH-UBS가 활성화된 MAPK와 NF-κB 신호전달체계를 억제하여 염증매개물질(NO, PGE2, IL-6, TNF-α) 및 MMPs(MMP-1, 3, 13)를 억제하고 연골 기질(aggrecan, collagen) 분해를 억제하여 항골관절염 효능을 발휘함을 나타내며, 이는 FJH-UBS를 관절 및 연골 건강 개선제로의 활용 가능성을 제시한다. Boswellia serrata (BS) is widely employed for the treatment of several diseases such as arthritis, rhinitis, asthma, and several cancers. The present study investigates the anti-osteoarthritis activity and the underlying mechanism of the ethanol extract of BS gum resin (FJH-UBS) enriched with keto-β-boswellic acid and 3-O-acetyl-11-keto-β-boswellic acid compared to the conventional BS extract by the additional process of oil removal with hexane. An in vitro osteoarthritis-like model was established using interleukin (IL)-1β-stimulated human SW1353 chondrocytes. The SW1353 cells were stimulated with IL-1β (10 ng/mL) and treated with FJH-UBS (0∼20 ㎍/mL) for 24 h. FJH-UBS reversed the IL-1β-induced increase in the protein and mRNA expressions of nitric oxide/inducible nitric oxide synthase, prostaglandin E2/cyclooxygenase, IL-6, tumor necrosis factor-α, matrix metalloproteinase (MMP)-1, MMP-3, and MMP-13, and reversed the IL-1β-induced downregulation of aggrecan and type II collagen. In addition, FJH-UBS reversed the IL-1β-induced increases in p65 nuclear factor-κB (NF-κB), inhibitor-κ-Bα, and the mitogen-activated protein kinase (MAPK) family (extracellular signaling-regulated kinase, p38, c-jun-N-terminal-kinase) phosphorylation, suggesting an anti-inflammatory activity mediated by blocking these key signaling transduction pathways. These results indicate that FJH-UBS is a potential therapeutic agent for osteoarthritis, exerting its effect via inhibition of the IL-1β-induced inflammation and inhibiting extracellular matrix degradation by suppressing the NF-κB and MAPK pathways.