A Novel Small-Molecule Inhibitor Targeting the IL-6 Receptor β Subunit, Glycoprotein 130

Hong, Soon-Sun,Choi, Jung Ho,Lee, Sung Yoon,Park, Yeon-Hwa,Park, Kyung-Yeon,Lee, Joo Young,Kim, Juyoung,Gajulapati, Veeraswamy,Goo, Ja-Il,Singh, Sarbjit,Lee, Kyeong,Kim, Young-Kook,Im, So Hee,Ahn, Sun The American Association of Immunologists, Inc. 2015 JOURNAL OF IMMUNOLOGY Vol.195 No.1

<P>IL-6 is a major causative factor of inflammatory disease. Although IL-6 and its signaling pathways are promising targets, orally available small-molecule drugs specific for IL-6 have not been developed. To discover IL-6 antagonists, we screened our in-house chemical library and identified-LMT-28, a novel synthetic compound, as a candidate IL-6 blocker. The activity, mechanism of action, and direct molecular target of LMT-28 were investigated. A reporter gene assay showed that LMT-28 suppressed activation of STAT3 induced by IL-6, but not activation induced by leukemia inhibitory factor. In addition, LMT-28 downregulated IL-6-stimulated phosphorylation of STAT3, gp130, and JAK2 protein and substantially inhibited IL-6-dependent TF-1 cell proliferation. LMT-28 antagonized IL-6-induced TNF-alpha production in vivo. In pathologic models, oral administration of LMT-28 alleviated collagen-induced arthritis and acute pancreatitis in mice. Based on the observation of upstream IL-6 signal inhibition by LMT-28, we hypothesized IL-6, IL-6R alpha, or gp130 to be putative molecular targets. We subsequently demonstrated direct interaction of LMT-28 with gp130 and specific reduction of IL-6/IL-6R alpha complex binding to gp130 in the presence of LMT-28, which was measured by surface plasmon resonance analysis. Taken together, our data suggest that LMT-28 is a novel synthetic IL-6 inhibitor that functions through direct binding to gp130.</P>

Interferon-γ Inhibits in vitro Mobilization of Eosinophils by Interleukin-5

Park, Choon-Sik,Choi, Eun Nam,Kim, Jung Sun,Choi, Yun Sung,Rhim, Tai Youn,Chang, Hun Soo,Chung, Il Yup S. Karger AG 2005 International archives of allergy and immunology Vol.136 No.3

<P><I>Background:</I> Th2 cytokines play pivotal roles in allergic inflammation, including eosinophilia, and their actions are antagonized by Th1 cytokines, conferring them therapeutic potential. <I>Methods:</I> In this study, we examined the ability of a number of cytokines to suppress the activation of eosinophils that function as effector cells for allergic airway diseases. <I>Results:</I> Interleukin (IL)-5, IL-6, and tumor necrosis factor (TNF) induced an eosinophil shape change, whereas interferon (IFN)-γ significantly inhibited the shape change. Other cytokines, including IL-1β, IL-4, IL-10 and IL-13, had little or only slightly enhancing or reducing effects on the shape change. We further analyzed the IFN-γ effect, showing that pretreatment with IFN-γ strongly suppressed IL-5-induced eosinophil shape change, and cycloheximide (CHX) abrogated the suppression by IFN-γ, suggesting that new protein synthesis is required for the inhibitory effect by this cytokine. In agreement with these results, IFN-γ blocked the eosinophil migration and ERK phophorylation induced by IL-5, and the addition of CHX restored eosinophil chemotaxis. <I>Conclusions:</I> Collectively, IFN-γ may attenuate eosinophilic inflammation by directly negating eosinophil mobilization.</P><P>Copyright © 2005 S. Karger AG, Basel</P>

OMC-2010 추출물이 마우스의 비장세포 cytokine 생성에 미치는 영향

배기상 ( Gi Sang Bae ),박경철 ( Kyoung Chel Park ),최선복 ( Sun Bok Choi ),조일주 ( Il Joo Jo ),서상완 ( Sang Wan Seo ),김종진 ( Jong Jin Kim ),신용국 ( Yong Kook Shin ),김민선 ( Min Sun Kim ),박규환 ( Kyu Hwan Park ),김현식 ( Hyu 대한본초학회 2012 大韓本草學會誌 Vol.27 No.5

Objective : This study was performed to estimate the effects of OMC-2010 extract on cytokine production in mouse spleen cells. Methods : Mouse spleen cells were pre-treated with ethanol and water extract of OMC-2010 for 1 h, then stimulated with lipopolysaccharide (LPS, 1 μg/ml) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokines. Results : OMC-2010 ethanol extract significantly inhibited the LPS-induced interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and IL-5 mRNA expressions, but not shown such changes in IL-6, IL-4, IL-13. OMC-2010 water extract significantly inhibited the LPS-induced TNF-alpha, and IL-5 mRNA expressions, but not shown such changes in IL-1beta, IL-6, IL-4, IL-13. Conclusions : Theses results could suggest that both ethanol and water OMC-2010 extract could inhibit the TNF-alpha and IL-5 mRNA expression.

Chronic alcohol consumption induces migration of IL-1R2+ monocytes from the bone marrow into the liver by neuro-immunologic pathway

Young-Ri Shim,Hee-Hoon Kim,Keungmo Yang,Tom Ryu,Kyurae Kim,Sung Eun Choi,Minjeong Kim,Chae-Rin Woo,Young-Sun Lee,Won-Il Jeong 한국실험동물학회 2021 한국실험동물학회 학술발표대회 논문집 Vol.2021 No.7

Liver is challenged by diverse detrimental substances through multiple metabolic processes, but it is less prone to inflammation. In chronic alcohol consumption, although the migration of monocytes from bone marrow (BM) into liver is increased, alcoholic hepatitis rarely occurs. Thus, we investigated the sub-population of liver macrophages showing anti-inflammatory roles through single-cell RNA sequencing (scRNA-Seq) after chronic EtOH-feeding. Interestingly, in scRNA-seq and flow cytometry analyses of hepatic macrophages, the phenotype of Ly6Clow (anti-inflammatory) cells was dramatically altered by ethanol intake. In particular, they were highly expressed interleukin-1 type II receptor (IL-1R2), a decoy receptor of IL-1β. Intriguingly, IL-1R2+ Ly6Clow macrophages showed decreased CX3CR1 expression, which was confirmed not only in the liver, but also BM and blood, suggesting monocytes from BM affected by ethanol might migrate into the liver. We found that the Leptin Receptor+ mesenchymal stromal cells (LepR+ MSCs), which were located around blood vessels expressing CX3CL1 to hold CX3CR1+ macrophages, could express alcohol dehydrogenase to metabolize ethanol in BM. Ethanol metabolism in LepR+ MSCs was induced both production of chemokines (CXCL9 and 10) and the excretion of glutamate via cystine-glutamate anti-porter xCT to recruit and activate the CXCR3+ BM NK cells to produce interferon-γ in a metabotropic glutamate receptor 5 (mGluR5)-dependent manner. Indeed, IFN-γ production was significantly decreased in EtOH-fed mice when we depleted mGluR5 in NK cells. In turn, NK cell-derived IFN-γ down-regulated CX3CR1 expression in BM Ly6Clow monocytes, consequently induced egress of Ly6Clow monocytes into the blood and migration into the liver to suppress alcoholic inflammation. In conclusion, glutamate of LepR+ MSCs imposed egress license on anti-inflammatory IL-1R2+ Ly6Clow monocytes through NK cell-derived IFN-γ-mediated suppression of CX3CR1, suggesting a potential therapeutic inter-organ crosstalk between BM and liver in alcoholic liver disease.

Anti-photoaging effect of fermented agricultural by-products on ultraviolet B-irradiated hairless mouse skin

CHOI, SUN-IL,JUNG, TAE-DONG,CHO, BONG-YEON,CHOI, SEUNG-HYUN,SIM, WAN-SUP,HAN, XIONGGAO,LEE, SANG JONG,KIM, YOUNG-CHEUL,LEE, OK-HWAN UNKNOWN 2019 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.44 No.2

<P> Processed products from agricultural produce generate a large number of agricultural by-products that contain a number of functional substances. These are often discarded owing to the lack of suitable processing methods. The present study investigated the anti-photoaging properties of fermented rice bran (FRB), soybean cake (FSB) and sesame seed cake (FSC) on ultraviolet B (UVB)-irradiated hairless mouse skin. Results indicated that the oral administration of FRB, FSB and FSC effectively inhibited the UVB irradiation-induced expression of matrix metalloproteinase (MMP)-2, MMP-9, MMP-3 and MMP-13. Reverse transcription-quantitative polymerase chain reaction results also demonstrated that FRB, FSB and FSC significantly inhibited the UVB-induced expression of the genes encoding tumor necrosis factor-α, inducible nitric oxide synthase, interleukin (IL)-6 and IL-1β when compared with the UVB-vehicle group (P<0.05). Additionally, collagen degradation and mast cell infiltration were reduced in hairless mouse skin. Furthermore, UVB-induced wrinkle formation was also significantly reduced in mouse skin compared with the UVB-vehicle group (P<0.05). These results reveal that fermented agricultural by-products may serve as potential functional materials with anti-photoaging activities. </P>

Expression of cytokine genes and increased nuclear factor-kappa B activity in the brains of scrapie-infected mice

Choi, Eun-Kyoung,Kim, Jae-Il,Ju, Won-Kyu,Carp, Richard I.,Wisniewski, Henryk M.,Kim, Jin,Choi, Jin-Ho,Kim, Yong-Sun 한림대학교 부설 환경ㆍ생명과학연구소 1999 일송 의학ㆍ생명과학 심포지엄 Vol.- No.1

A number of aspects of the pathogenesis of scrapie remain to be elucidated. The cellular and molecular aspects of the neuropathology in scrapte suggest the possibility that the proinflammatory cytokines could act as apthogenic mediators in this neurodegencrative disease. To understand this possibility, we examined the expression of prounflammatory cytokine genes in brains of IM mice-infected with 87V scrapie agent. Additionally, we also analyzed the activity of nuclear factor-kappa B (NF-k B), which is the major transcriptional activator for inflammatory cylokinesk and formation of reactive oxygen species (ROS) as a common upsiteam messenger for its activation. The induction of mRNAs of the inflammatory cytokines, IL-1α, IL-1β and TNF-α, was detected only int he brains of scrapie-infected mice. The activity of NF-kB was significantly increased in the nuclear extracts from brains of the scrapie-infected group and the immunoreactivity of NF-kB was increased in the hippocampus and thalamus in the brains of scrapie-infected mice. The NF-kB immunoreactivity was observed mainly in GFAP-positive astrocytes and also detected int he PrP-amyloid plaques in the brains of 87V scrapie-infected mice Gene expression of IL-6 and NOS, the representative target genes for NF-kB activation, wee activated only in the infected group. The production of LOS was significantly increased in the brain mitochondrial litions of scrapie-infected mice. These results suggest that prion accumulation in astrocytes might activate NF-kB through the increase of ROS generation, and thus alterations in NF-kB-directed gene expression may contribute to both the neurodegeneration and proinflammatory responses which occur in scrapie C 1999 Elsevier Science B. V. All rights reserved.

Structure and Immune Modulating Activity of a Polysaccharide Purified from Mulberry Fruits Pericarp

Ji Sun Lee,Andriy Synytsya,Hyun Bok Kim,Woo Jung Kim,Seong Jae Jang,Mi Ja Chung,Ju Hee Ko,Doo Jin Choi,Ha Na Choi,Yong Il Park 한국당과학회 2011 한국당과학회 학술대회 Vol.2011 No.1

In the present study, we investigated structure and immune modulating activity of a polysaccharide (tentatively named JS-MP-1), previously purified from Mulberry fruits (Morus bombycis Koidz) pericarp. Based on HPAEC-PAD analysis, major monosaccharides of JS-MP-1 were shown to be galactose (37.6% in mole percentage), arabinose (36.3%) and rhamnose (18.4%), respectively. EA analysis did not show the presence of sulfur (0.07%). Biochemical analysis showed neutral sugars (35.25%), uronic acid (11.25%), sulfate (0.16%) and proteins (0.3%). FT-IR spectra of JS-MP1 confirmed the presence of carboxyl groups (uronic acid – α-GalAp in RG1) and COC, CO and CC stretching vibrations at 948 – 1150 cm-1 corresponding to polysaccharides. Peak at 894 cm-1 indicates the presence of β-Galp units. 1H-NMR spectrum (in D2O) confirmed the presence of α-1,2-Rhap, α-Araf, β -Galp and α-1,4-GalAp units. The most intense HMQS signals were observed for terminal α-Araf. The presence of methylester groups in the residues of α -1,4-GalAp was confirmed by the HMQS peak at C 54.3/H 4.01 ppm (OCH3). Taken collectively, the JS-MP-1 is probably a rhamnogalacturonan type 1 (RG1), which contains: (a) partially methyl esterified α-1,4-D-GalAp and α -1,2-L-Rhap units in backbone, and (b) α-L-Araf and β-Galp are bound to Rha as terminal units or more complex side chains (arabinan, galactan and/or arabinogalactan). The ELISA assay showed that JS-MP-1 remarkably stimulates the release of the proinflammatory cytokines, TNF-α and IL-6, in a dose-dependent manner, from the RAW 264.7 cells. RT-PCR analysis also showed a significant induction of the iNOS and COX-2 gene expression. These results suggest that JS-MP-1 isolated from the Mulberry fruits has immune modulating activities.

(무기환)戊己丸의 RAW 264.7 세포에 대한 항염증작용 연구

김일현 ( Il Hyun Kim ),최종환 ( Chong Hwan Choi ),이세원 ( Se Won Lee ),송용선 ( Yung Sun Song ) 한방재활의학과학회 2013 한방재활의학과학회지 Vol.23 No.3

ObjectivesThe aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. MethodsCells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines (PGE2, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. ResultsMGH inhibited the production of NO, PGE2, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. ConclusionsThese results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes. (J Korean Med Rehab 2013;23(3):27-35) ObjectivesThe aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. MethodsCells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines (PGE2, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. ResultsMGH inhibited the production of NO, PGE2, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. ConclusionsThese results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes. (J Korean Med Rehab 2013;23(3):27-35)

LPS로 유도한 RAW 264.7 세포의 염증반응에서 자초(紫草)의 항염증 효과

최선복 ( Sun Bok Choi ),배기상 ( Gi Sang Bae ),조일주 ( Il Joo Jo ),박경철 ( Kyoung Chel Park ),서승희 ( Seung Hee Seo ),김동구 ( Dong Goo Kim ),신준연 ( Joon Yeon Shin ),곽태신 ( Tae Sin Gwak ),이정현 ( Jung Hyun Lee ),이금산 ( G 대한본초학회 2013 大韓本草學會誌 Vol.28 No.2

Objective: Lithospermum Erythrorhizon (LE) has been used as an anti-bacterial and anti-inflammatory agent. However, it is unclear that LE aqueous extract could show the anti-inflammatory effects in RAW 264.7cells. The purpose of this study was to investigate the anti-inflammatory effect of aqueous extract from LE on lipopolysaccharide (LPS) - induced inflammatory response. Methods: To measure out the cytotoxicity of LE, we performed the MTT assay. To evaluate the anti-inflammatory effects of LE, we examined the inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-a, interleukin, (IL)-1β and (IL)-6) on RAW 264.7 cells. We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-B (NF-κB) activation by western blot. Results : Aqueous Extract from LE itself did not have any cytotoxic effect in RAW 264.7 cells. Aqueous extract from LE inhibited LPS-induced productions of inflammatory mediators such as NO, PGE2, and pro-inflammatory cytokines including TNF-a, IL-1β and IL-6 in RAW 264.7cells. In addition, LE inhibited the phosphorylation of p38 kinases (p38), c-Jun NH2-terminal kinase (JNK), and NF-κB activation in RAW 264.7 cells. Conclusion : LE down-regulated LPS-induced production of inflammatory mediators through the inhibition of p38, JNK and NF-κB activation. Taken together, these results could provide the evidence for the anti-inflammatory effects of LE. Therefore, LE may be a novel target in the management of inflammation and help to support a potential strategy for prevention and therapy of inflammatory diseases.

OMC-2010 구성약재 배합추출물 투여가 Ovalbumin으로 유도한 마우스 알레르기성 기관지 천식에 미치는 영향

조일주 ( Il Joo Jo ),배기상 ( Gi Sang Bae ),최선복 ( Sun Bok Choi ),송호준 ( Ho Joon Song ),박성주 ( Sung Joo Park ),서상완 ( Sang Wan Seo ),옥주안 ( Joo An Ok ),김민선 ( Min Sun Kim ),백선종 ( Sun Jong Baek ),배익현 ( Ik Hyun Bae 대한본초학회 2013 大韓本草學會誌 Vol.28 No.5

Objectives: We recently have reported that constituents of OMC-2010 have an immuno-modulatory effects via inhibiting tumor necrosis factor (TNF)-alpha and interleukin (IL)-5. In this study, based on previous data, we investigated the effects of combinations with each OMC constituents on splenocyte cytotoxicity, cytokine productions, and ovalbumin (OVA) induced experimental allergic asthma. Methods: Mouse splenocytes were pre-treated with ethanol extract of constituents of Rehmannia glutinosa (RG), Pinellia ternata (PT), Schisandra chinensis (SC). We made 4 combinations using RG, PT, and SC (A;1:1:1, B;2:1:1, C;1:2:1, D;1:1:2). The cells were pretreated with A, B, C, or D for 1 h, then stimulated with lipopolysaccharide (LPS, 1 ㎍/ml) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokine productions. Then using effective combination from RG, PR and SC, we administrated the combination orally, then challenged with OVA to induce asthma. Then we analyzed the airway hyper-reactivity (AHR), lung histology and lung TNF-α and IL-5 mRNA. Results: A. B. C. and D did not showed significant cytotoxicity on splenocytes. Pre-treatment of A inhibited the expression of TNF-α and IL-5 significantly, but not B, C, and D. In experimental asthma, administration of A significantly inhibited the increase of AHR, lung damage, TNF-α and IL-5 expression. Conclusions: Theses results could suggest that inhibitory effects of the ideal combination with RG, PT and SC (1:1:1) could be applied to treatment of asthma and study of asthma mechanisms.