Inducible Nitric Oxide Synthase Inhibitor form Mela azedarach var. Japonica

Kwon, Hak-Cheol,Lee, Byeong-Gon,Kim, Seung-Hee,Jung, Chil-Mann,Hong, Sung-Youl,Han, Jeung-Whan,Lee, Hyang-Woo,Zee, Ok-Pyo,Lee, Kang-Ro The Pharmaceutical Society of Korea 1999 Archives of Pharmacal Research Vol.22 No.4

In bioassay-guided search for inducible nitric oxide synthase (iNOS) inhibitory compounds from higher plants of South Korea, two $\beta$-carboline (2) have been isolated form the cortex of Melia azedarach var. japonica. The structures of these compounds were elucidated on the basis of spectroscopic data. Compounds 1 to 2 showed marked inhibitory activity of iNOS on LPS-and interferon-${\gamma}$-stimulated RAW 264.7 cells.

Inducible Nitric Oxide Synthase Inhibitors from Melia azedarach var.Japonica

Kwon, Hak Cheol,Lee, Byeong Gon,Kim, Seung Hee,Jung, Chil Mann,Hong, Sung Youl,Han, Jeung Whan,Lee, Hyang Woo,Zee, Ok Pyo,Lee, Kang Ro 성균관대학교 약학연구소 1999 成均藥硏論文集 Vol.11 No.-

In bioassay-guided search for inducible nitric oxide synthase (iNOS) inhibitory compounds from higher plants of South Korea, two β-carboline alkaloids, 4-methoxy-1-vinyl-β-carboline (1) and 4,8-dimethoxy-1-vinyl-β-carboline (2) have been isolated from the cortex of Melia azedarach var. japonica. The structures of these compounds were elucidated on the basis of spectroscopic data. Compounds 1 and 2 showed marked inhibitory activity of iNOS on LPS- and interferon-γ-stimulated RAW 264.7 cells.

톨루엔 취급 근로자의 ALDH2 Genetic Polymorphism에 따른 뇨중 마뇨산 배설량

김창윤,정종학,권오춘,김성용,이중정,주리 大韓産業醫學會 1997 대한직업환경의학회지 Vol.9 No.2

In this study we evaluated the effects of the genetic polymorphism of aldehyde dehydrogenase2 (ALDH2) on the toluene metabolism and determined biological exposure indices(BEIs) for toluene by the genotypes of ALDH2. The study subject were 77 men workers who are handling toluene in a video tape manufacturing factory and a textile company. Through the face-to-face interview, the information about smoking and drinking behavior was obtained. For determination of ALDH2 polymorphism, 5ml of venous blood sample was obtained from each subject after informed consent. DNA was extracted from the buffy coat and ALDH2 genotyping were performed using a polymerase chain reaction(PCR) method. The genotype of ALDH2 was classified into the homozygous genotype of normal ALDH2(NN), and the heterozygous genotype of normal and inactive ALDH2(ND), and homozygous genotype of an inactive ALDH2(DD). The concentration of hippuric acid (HA), the main metabolite of toluene, was determined in urine specimens collected at the end of shift, corrected with creatinine(HA/C), and compared with BEI for toluene, which is 2.5 g/g creatinine. The personal exposure level of toluene were monitored, using personal air sampler and analyzed by gas chromatography. The frequencies of the three genotypes in this study subject were, NN : 45 (58.4%), ND : 26 (33.8%) and DD : 6 (7.8%), and frequencies of the genotypes in the middle or heavy toluene exposure workers were not significantly different from those in the mild toluene exposure workers. The frequencies of the DD type of ALDH2 was lower among alcohol drinkers than among non-drinkers. The urinary HA concentration of DD group was significantly lower than that of the NN or ND group, which suggests that the HA formation from toluene decreased in DD group. Regression lines were used to estimate the BEIs of the NN, ND, and DD groups. NN : y = 0.0085x + 0.23, r = 0.90 ND : y = 0.0074x + 0.21, r = 0.85 DD : y = 0.0041x + 0.82, r = 0.83 The three regression lines revealed that the estimated urinary HA concentration of NN, ND, and DD groups at 377 mg/㎥ toluene (TLV-TWA) exposure were 3.43, 3.00, and 2.37g/g than that of the ACGIH BEI (2.5 g/g creatinine) ; however, the HA level of DD group was lower than the BEI. These results suggest that the ACGIH BEI is not adequate to estimate the toluene exposure of the NN, ND and DD groups at the same time. Based upon these results, a new BEI for ALDH2 deficient workers may be necessary.

백합 절화시 지상부 잔존량이 구근의 비대에 미치는 영향

최종진(Choi Jong Jin),권경학(Kwon Kyeong Hak),서정근(Suh Jeung Keun) 한국원예학회 1997 한국원예학회 학술발표요지 Vol.15 No.1

Mizoribine Inhibits Production of Pro-inflammatory Cytokines and $PGE_2$ in Macrophages

Han, Shin-Ha,Kim, Kwang-Hee,Kim, Hyun-Yul,Kwon, Jeung-Hak,Han, Nam-Joo,Lee, Chong-Kil,Kim, Kyung-Jae The Korean Association of Immunobiologists 2007 Immune Network Vol.7 No.1

Background: Mizoribine (MZR) is an imidazole nucleoside isolated from Eupenicillium brefeldianum. MZR is currendy in clinical use for patients who have undergone renal transplantation. Therapeutic efficacy of MZR has also been demonstrated in rheumatoid arthritis and lupus nephritis. MZR has been shown to inhibit the proliferation or lymphocytes by interfering with inosine monophosphate dehydrogenase. Since the exact mechanism by which MZR benefits rheumatoid arthritis (RA) is not clear, we investigated the ability of MZR to direct its immunosuppressive influences on other antigen presenting cells, such as macrophages. Methods: Mouse macrophage RAW264.7 cells were stimulated with lipopolysaccharide in the presence of MZR. To elucidate the mechanism of the therapeutic efficacy in chronic inflammatory diseases, we examined the effects of MZR on the production of pro-inflammatory cytokines, nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ in macrophages. Results: MZR dose-dependendy decreased the production of nitric oxide and pro- inflammatory cytokines such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukins $1{\beta}$ (IL-${\beta}$ and IL-6 $PGE_2$. Examination of gene expression levels showed that the anti-inflammatory effect correlated with the down-regulation of inducible nitiric oxide synthase expression, cycloxygenase-2 expression and TNF-${\alpha}$ gene expression. Conclusion: In this work, we resulted whether MZR $(1.25{\sim}10{\mu}g/ml)$ inhibited macrophage activation by inhibiting secretion of pro-inflammatory cytokines, NO and $PGE_2$. These findings provide an explanation for the therapeutic efficacy of MZR in chronic inflammation-associated diseases.

Mizoribine Inhibits Production of Pro-inflammatory Cytokines and PGE₂ in Macrophages

Han, Shin-Ha,Kim, Kwang-Hee,Kim, Hyun-Yul,Kwon, Jeung-Hak,Han, Nam-Joo,Lee, Chong-Kil,Kim, Kyung-Jae 대한면역학회 2007 Immune Network Vol.7 No.1

Mizoribine (MZR) is an imidazole nucleoside isolated from Eupenicillium brefeldianum. MZR is currendy in clinical use for patients who have undergone renal transplantation. Therapeutic efficacy of MZR has also been demonstrated in rheumatoid arthritis and lupus nephritis. MZR has been shown to inhibit the proliferation or lymphocytes by interfering with inosine monophosphate dehydrogenase. Since the exact mechanism by which MZR benefits rheumatoid arthritis (RA) is not clear, we investigated the ability of MZR to direct its immunosuppressive influences on other antigen presenting cells, such as macrophages. Methods: Mouse macrophage RAW264.7 cells were stimulated with lipopolysaccharide in the presence of MZR. To elucidate the mechanism of the therapeutic efficacy in chronic inflammatory diseases, we examined the effects of MZR on the production of pro-inflammatory cytokines, nitric oxide (NO) and prostaglandin E₂(PGE₂) in macrophages. Results: MZR dose-dependendy decreased the production of nitric oxide and pro- inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukins 1β (IL β and IL-6 PGE₂. Examination of gene expression levels showed that the anti-inflammatory effect correlated with the down-regulation of inducible nitiric oxide synthase expression, cycloxygenase-2 expression and TNF-α gene expression. Conclusion: In this work, we resulted whether MZR (1.25∼10μg/ml) inhibited macrophage activation by inhibiting secretion of pro-inflammatory cytokines, NO and PGE₂. These findings provide an explanation for the therapeutic efficacy of MZR in chronic inflammation-associated diseases.

Auranofin, an Immunosuppressive Drug, Inhibits MHC Class I and MHC Class II Pathways of Antigen Presentation in Dendritic Cells

Han, Shin-Ha,Kim, Kwang-Hee,Song, Young-Cheon,Kim, Hyun-Yul,Kwon, Jeung-Hak,Lee, Young-Hee,Lee, Chong-Kil,Lee, Sang-Jin,Ha, Nam-Joo,Kim, Kyung-Jae 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.3

Auranofin (AF), an orally administered, gold-based, anti-arthritic agent, has emerged as a clinically useful therapeutic drug for the treatment of rheumatoid arthritis. In the present study, we examined the effects of AF on major histocompatibility complex (MHC)-restricted antigen presentation in dendritic cells (DCs), which are the most important accessory cells for the induction of T cell responses. A mouse dendritic cell line, DC2.4 cells, and DCs that were generated from mouse bone marrow cells by culturing with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4 were each pretreated with AF for 2 hr, and then incubated with ovalbumin (OVA). After the 2-hr incubation, the DCs were fixed, and the amounts of OVA $peptide-H-2K^b$ complexes were assessed using OVA-specific $CD8_+$ T cells. AF inhibited MHC class I-restricted presentation of exogenous OVA. This inhibitory activity of AF appeared to be due not only to the inhibition of the phagocytic activity of DCs, but also to the suppression of MHC molecule expression on DCs. AF also inhibited MHC class II-restricted presentation of exogenous OVA. These results show that AF exerts immunosuppressive activity at least in part by inhibiting MHC-restricted antigen presentation in professional antigen-presenting cells.

Auranofin Inhibits Overproduction of Pro-Inflammatory Cytokines, Cyclooxygenase Expression and $PGE_2$ Production in Macrophages

Han, Shin-Ha,Kim, Kwang-Hee,Kim, Hyun-Yul,Kwon, Jeung-Hak,Lee, Young-Hee,Lee, Chong-Kil,Song, Young-Cheon,Lee, Sang-Jin,Ha, Nam-Joo,Kim, Kyung-Jae 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.1

Auranofin (AF), a gold compound, is an orally active therapeutic agent used to treat rheumatoid arthritis (RA), a self-perpetuating inflammatory disease. RA is characterized by autoimmune-mediated proliferation of synovial cells that leads to inflammation, pain, and swelling in most major joints. However, the mechanism as to how AF relieves RA symptoms has not been fully elucidated. The object of this study was to examine the ability of AF to immunomodulate macrophages as antigen presenting cells (APCs). Macrophages are recognized as playing an important role in the pathogenesis of RA, in that there is a relative abundance of macrophagederived cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), $interleukin-1{\beta}$ ($IL-1{\beta}$) and interleukin-6 (IL-6) in rheumatoid synovium. In this work, we tested whether AF (2.5-20 mM) could inhibit inflammatory activity in the macrophage cell line RAW 264.7. AF decreased production of nitric oxide (NO) and the pro-inflammatory cytokines, $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 in macrophages. Furthermore, AF inhibited cyclooxygenase-2 (COX-2)-dependent prostaglandin E2 (PGE2) production in a concentration-dependent manner. In conclusion, these findings may provide an explanation for the clinical effects of AF in patients with RA.

Effects of Mizoribine on MHC-Restricted Exogenous Antigen Presentation in Dendritic Cells

Song, Young-Cheon,Han, Shin-Ha,Kim, Hyun-Yul,Kim, Kwang-Hee,Kwon, Jeung-Hak,Lee, Sang-Jin,Ha, Nam-Joo,Lee, Young-Hee,Lee, Chong-Kil,Kim, Kyung-Jae The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.12

Mizoribine (MZR) has been shown to possess immunosuppressive activity that selectively inhibits the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. The efficacy of MZR is not only in patients who have had renal transplantation, but also in patients with rheumatoid arthritis (RA), lupus nephritis, and primary nephritic syndrome. Because the exact mechanism of its immunosuppressive action is not clear, the object of this study was to examine the ability of MZR to regulate the antigen presenting cells (APCs), dendritic cells (DCs). In this work, we tested whether MZR ($1{\sim}10\;{\mu}g/mL$) could inhibit the cross-presentation of DCs. DC2.4 cells ($H-2K^{b}$) or bone marrow-derived DCs (BM-DCs) generated from BM cells of C57BL/6 mouse ($H-2K^{b}$) were cultured in the presence of MZR with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by a T cell hybridoma, B3Z, that recognizes OVA (257-264 : SIINFEKL)-$H-2K^{b}$ complex and expresses-galactosidase. MZR profoundly inhibited the expression of SIINFEKL-$H-2K^{b}$ complexes. This inhibitory activity of MZR appeared to affect the phagocytic activity of DCs. MZR also decreased IL-2 production when we examined the effects of MZR on $CD4^{+}$ T cells. These results provide an understanding of the mechanism of immunosuppressive activity of MZR on the inhibition of MHC-restricted antigen presentation and phagocytic activity in relation to their actions on APCs.

Activation of Macrophages by the Components Produced from Cordyceps militaris

Kim, Hyun-Yul,Kim, Kwang-Hee,Han, Shin-Ha,Lee, Seong-Jung,Kwon, Jeung-Hak,Lee, Sung-Won,Kim, Kyung-Jae The Korean Association of Immunobiologists 2007 Immune Network Vol.7 No.2

Background: Cordyceps militaris have been reported to modify the immune and inflammatory responses both in vivo and in vitro. Macrophages play important roles in the innate immunity through the phagocytosis of antigens. This study examined the effects of Cordyceps militaris on the activation of murine macrophage RAW 264.7 cells and primary macrophages. Methods: The components contained in culture broth of Cordyceps militaris were purified by propyl alcohol extraction and HP 20 column chromatography to CMDB, CMDBW, CMDB5P, and CMDB25P. The amounts of nitric oxide (NO) were determined by using ELISA, Griess reagent respectively. The amounts of some cytokines were determined by using ELISA, western blot, and RT-PCR The expression levels of cell surface molecules (ICAM-1, B7-1 and B7-2) were measured by flow cytometric analysis. Results: All the components of Cordyceps militaris produced significant amounts of NO. In particular, CMDB produced much more NO in RAW 264.7 cells and primary macrophages than other fractions of Cordyceps militaris. CMDB increased significantly the production of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-1${\beta}$, and IL-6 dose-dependently in RAW 264.7 cells. Examination of the gene expression level also showed that the enhanced production of cytokines was correlated with the up-regulation of i-NOS expression, cycloxygenase (COX)-2 expression, IL-1${\beta}$ and IL-6 expression, and TNF-${\alpha}$ expression on the expression of mRNAs by semi-quantitative RT-PCR Western blot analysis also confirmed that CMDB enhances the expression level of these cytokines. Conclusion: These results show that CMDB stimulates the production of NO and pro-inflammatory cytokines and can also up-regulate the gene expression levels in macrophages.