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Shin, Seulmee,Kim, Bong-Yoon,Jeon, Hyung-Yook,Lee, Aeri,Lee, Sungwon,Sung, Seung-Hyun,Park, Chan-Su,Lee, Chong-Kil,Kong, Hyunseok,Song, Youngcheon,Kim, Kyungjae 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.5
Adiponectin is an adipocyte hormone involved in glucose and lipid metabolism. The aim of this study was to develop a human adiponectin expression system in transgenic silkworm using a human adiponectin expression vector. The silk gland of the silkworm is a highly specialized organ that has the wonderful ability to synthesize and secrete silk protein. To express human adiponectin in the silk gland of transgenic silkworm, targeting vectors pB-A3-adiponectin-IRES-RFP and pB-Ser1-adiponectin-IRES-RFP were constructed and then introduced into the silkworm pupa. The transgenic silkworms were verified by PCR and then generated. The level of adiponectin in the transgenic silkworm was 6-10 ng/50 mg of freeze-dried powder, and western blotting using an antibody against human adiponectin demonstrated a specific band with a molecular weight of 30 kDa in the silkworm. These results showed that human adiponectin introduced into the silkworm genome was expressed successfully on a large-scale.
Shin, Seulmee,Park, Yoonhee,Kim, Seulah,Oh, Hee-Eun,Ko, Young-Wook,Han, Shinha,Lee, Seungjeong,Lee, Chong-Kil,Cho, Kyunghae,Kim, Kyungjae The Korean Association of Immunobiologists 2010 Immune Network Vol.10 No.4
Background: Cordyceps militarys water extract (CME) has been reported to exert antitumor and immunomodulatory activities in vivo and in vitro. However, the therapeutic mechanism has not yet been elucidated. In this study, we examined the effects of CME on the antigen presenting function of antigen presenting cells (APCs). Methods: Dendritic cells (DCs) were cultured in the presence of CME, and then allowed to phagocytose microspheres containing ovalbumin (OVA). After washing and fixing the efficacy of OVA, peptide presentation by DCs were evaluated using CD8 and CD4 T cells. Also, we confirmed the protein levels of proinflammatory cytokines through western blot analysis. Results: CME enhanced both MHC class I and class II-restricted presentation of OVA in DCs. In addition, the expression of both MHC class I and II molecules was enhanced, but there was no changes in the phagocytic activity of exogenous OVA. Furthermore, CME induced the protein levels of iNOS, COX-2, proinflammatory cytokines, and nuclear p65 in a concentration-dependent manner, as determined by western blot. Conclusion: These results provide an understanding of the mechanism of the immuno-enhancing activity of CME on the induction of MHC-restricted antigen presentation in relation to their actions on APCs.
신슬미,김경제,조경혜 서울여자대학교 자연과학연구소 2013 자연과학연구논문집 Vol.25 No.-
Cordyceps militaris (CM) has been used in traditional medicine to treat numerous diseases in vivo and in vitro. The molecular mechanism of CM pharmacological and biochemical actions on immune responses has not been clearly elucidated. In the present study, the water extract of CM (CME) and cordycepin (3’-deoxyadenosine) were prepared to examine the action mechanism of CM on immune response. Cordycepin, nucleoside analogue, is a metabolite of CM. This study demonstrated how CME or cordycepin itself regulates immune response by the production of pro-inflammatory cytokines ex-vivo. CME induced the expression of NO and pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α in primary macrophages at mRNA and protein levels. On the other hand, cordycepin inhibited pro-inflammatory mediators in LPS-stimulated macrophages in a concentration of between 5 µg to 40 µg. The present results showed that CME induces immune response via NF-κB activation in macrophages meanwhile cordycepin inhibits inflammation via suppression of NF-κB activation in activated macrophages by LPS. These results demonstrate that Cordyceps militaris (CM) modulates immune responses, and it may potentially be useful immunomodulating agent for immune deficiency disorders.
Hyun, Bobae,Shin, Seulmee,Lee, Aeri,Lee, Sungwon,Song, Youngcheon,Ha, Nam-Joo,Cho, Kyung-Hea,Kim, Kyungjae The Korean Association of Immunobiologists 2013 Immune Network Vol.13 No.4
Obesity is consistently increasing in prevalence and can trigger insulin resistance and type 2 diabetes. Many lines of evidence have shown that macrophages play a major role in inflammation associated with obesity. This study was conducted to determine metformin, a widely prescribed drug for type 2 diabetes, would regulate inflammation through down-regulation of scavenger receptors in macrophages from obesity-induced type 2 diabetes. RAW 264.7 cells and peritoneal macrophages were stimulated with LPS to induce inflammation, and C57BL/6N mice were fed a high-fat diet to generate obesity-induced type 2 diabetes mice. Metformin reduced the production of NO, $PGE_2$ and pro-inflammatory cytokines ($IL-1{\beta}$, IL-6 and $TNF-{\alpha}$) through down-regulation of $NF-{\kappa}B$ translocation in macrophages in a dose-dependent manner. On the other hand, the protein expressions of anti-inflammatory cytokines, IL-4 and IL-10, were enhanced or maintained by metformin. Also, metformin suppressed secretion of $TNF-{\alpha}$ and reduced the protein and mRNA expression of $TNF-{\alpha}$ in obese mice as well as in macrophages. The expression of scavenger receptors, CD36 and SR-A, were attenuated by metformin in macrophages and obese mice. These results suggest that metformin may attenuate inflammatory responses by suppressing the production of $TNF-{\alpha}$ and the expressions of scavenger receptors.
Metformin Down-regulates TNF-α Secretion via Suppression of Scavenger Receptors in Macrophages
Bobae Hyun,Seulmee Shin,Aeri Lee,이성원,송영천,하남주,Kyung-HeaCho,김경제 대한면역학회 2013 Immune Network Vol.13 No.4
Obesity is consistently increasing in prevalence and can trigger insulin resistance and type 2 diabetes. Many lines of evidence have shown that macrophages play a major role in inflammation associated with obesity. This study was conducted to determine metformin, a widely prescribed drug for type 2 diabetes, would regulate inflammation through downregulation of scavenger receptors in macrophages from obesity- induced type 2 diabetes. RAW 264.7 cells and peritoneal macrophages were stimulated with LPS to induce inflammation, and C57BL/6N mice were fed a high-fat diet to generate obesity-induced type 2 diabetes mice. Metformin reduced the production of NO, PGE2 and pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) through down-regulation of NF-κB translocation in macrophages in a dose-dependent manner. On the other hand, the protein expressions of anti- inflammatory cytokines, IL-4 and IL-10, were enhanced or maintained by metformin. Also, metformin suppressed secretion of TNF-αand reduced the protein and mRNA expression of TNF-α in obese mice as well as in macrophages. The expression of scavenger receptors, CD36 and SR-A, were attenuated by metformin in macrophages and obese mice. These results suggest that metformin may attenuate inflammatory responses by suppressing the production of TNF-α and the expressions of scavenger receptors.
Kim, Seulah,Shin, Seulmee,Hyun, Bobae,Kong, Hyunseok,Han, Shinha,Lee, Aeri,Lee, Seungjeong,Kim, Kyungjae 대한면역학회 2012 Immune Network Vol.12 No.5
Dioscoreae Rhizome (DR) has been used in traditional medicine to treat numerous diseases and is reported to have anti-diabetes and anti-tumor activities. To identify a bioactive traditional medicine with anti-inflammatory activity of a water extract of DR (EDR), we determined the mRNA and protein levels of proinflammatory cytokines in macrophages through RT-PCR and western blot analysis and performed a FACS analysis for measuring surface molecules. EDR dose-dependently decreased the production of NO and pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and PGE<sub>2</sub>, as well as mRNA levels of iNOS, COX-2, and pro-inflammatory cytokines, as determined by western blot and RT-PCR analysis, respectively. The expression of co-stimulatory molecules such as B7-1 and B7-2 was also reduced by EDR. Furthermore, activation of the nuclear transcription factor, NF-κB , but not that of IL-4 and IL-10, in macrophages was inhibited by EDR. These results show that EDR decreased pro-inflammatory cytokines via inhibition of NF-κB -dependent inflammatory protein level, suggesting that EDR could be a useful immunomodulatory agent for treating immunological diseases.
Kim, Seulah,Shin, Seulmee,Hyun, Bobae,Kong, Hyunseok,Han, Shinha,Lee, Aeri,Lee, Seungjeong,Kim, Kyungjae The Korean Association of Immunobiologists 2012 Immune Network Vol.12 No.5
Dioscoreae Rhizome (DR) has been used in traditional medicine to treat numerous diseases and is reported to have anti-diabetes and anti-tumor activities. To identify a bioactive traditional medicine with anti-inflammatory activity of a water extract of DR (EDR), we determined the mRNA and protein levels of proinflammatory cytokines in macrophages through RT-PCR and western blot analysis and performed a FACS analysis for measuring surface molecules. EDR dose-dependently decreased the production of NO and pro-inflammatory cytokines such as IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and $PGE_2$, as well as mRNA levels of iNOS, COX-2, and pro-inflammatory cytokines, as determined by western blot and RT-PCR analysis, respectively. The expression of co-stimulatory molecules such as B7-1 and B7-2 was also reduced by EDR. Furthermore, activation of the nuclear transcription factor, NF-${\kappa}B$, but not that of IL-4 and IL-10, in macrophages was inhibited by EDR. These results show that EDR decreased pro-inflammatory cytokines via inhibition of NF-${\kappa}B$-dependent inflammatory protein level, suggesting that EDR could be a useful immunomodulatory agent for treating immunological diseases.