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황금작약탕(黃芩芍藥湯)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구
김마룡 ( Ma Ryong Kim ),강옥화 ( Ok Hwa Kang ),김성배 ( Sung Bae Kim ),강희정 ( Hee Jung Kang ),김지은 ( Ji Eun Kim ),황형칠 ( Hyeong Chil Hwang ),김인원 ( In Won Kim ),권동렬 ( Dong Yeul Kwon ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.1
Objectives : Hwanggeumjakyak-tang (huangqin shaoyao tang, HJT) has been used to treat acute enteritis in traditional oriental medicine. However, there has been a lack of studies regarding the effects of HJT on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So we examined the effect of HJT water extract on pro-inflammatory mediators in lipopolysaccharide (LPS) - stimulated mouse macrophage, RAW 264.7 cells, Methods : Cells were treated with 2 ug/mL of LPS 1 h prior to the addition of HJT. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The expression of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) was investigated by Western blot, RT-PCR. The content of level of cytokines (prostaglandin (PG) E₂, interleukin (IL)-6, IL-12, Tumor necrosis factor-alpha (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit, Results : HJT inhibited the production of NO, PGE₂, IL-6 as well as the expressions of iNOS, COX-2 but did not inhibit the production of IL-12, TNF-α, MCP-1 in the murine macrophage, RAW 264.7 cells. HJT also had suppression effects of LPS- induced MAPKs activation, Conclusion : These results suggest that HJT has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.
현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구
김마룡 ( Ma-ryong Kim ),강옥화 ( Ok-hua Kang ),공룡 ( Ryong Kong ),서윤수 ( Yun-soo Seo ),주전 ( Tian Zhou ),김상아 ( Sang-a Kim ),김은수 ( Eun-su Kim ),신민아 ( Min-a Sin ),이영섭 ( Young-seob Lee ),권동렬 ( Dong-yeul Kwon ) 대한본초학회 2017 大韓本草學會誌 Vol.32 No.2
Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan (HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. PGE2 were measured using EIA kit. Proinflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. NF-κB/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, TNF-α, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of NF-κB in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of NF-κB delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced PGE2,NO, IL-6 and TNF-α
대식세포에서 수련환(茱連丸) 물추출물의 항염증작용에 관한 연구
윤여환 ( Yeo Hwan Yoon ),김성배 ( Sung Bae Kim ),강옥화 ( Ok Hwa Kang ),문수현 ( Su Hyun Mun ),서윤수 ( Yun Soo Seo ),양다운 ( Da Wun Yang ),강다혜 ( Da Hye Kang ),위경 ( Gyeong Wi ),임재수 ( Jae Soo Lim ),김마룡 ( Ma Ryong Kim ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6
Objectives : Suryeon-hwan (SRH) exhibits potent anti-inflammatory activity with an unknown mechanism. However, there has been a lack of studies regarding the effects of SRH on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So, the investigation focused on whether SRH inhibited nitric oxide (NO) and prostaglandin E2 (PGE2) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with 200 ng/mL of LPS 30 min prior to the addition of SRH. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The content of level of cytokines (PGE, IL-6) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. Results : We found that SRH inhibited LPS-induced NO, PGE2 and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, SRH suppressed the LPS-induced phosphorylation of MAPK and extracellular signal-regulated kinase 1/2 (ERK 1/2) activation. Conclusions : These results suggest that SRH has inhibitory effects on LPS-induced PGE2, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the phosphorylation of MAPKs following activation.
인간 비만세포에서 PMA와 A23187에 의해 유도된 전염증 매개체에 대한 신효월도산 추출물의 항염증 효과
위경 ( Gyeong Wi ),양다운 ( Da Wun Yang ),강옥화 ( Ok Hwa Kang ),김성배 ( Sung Bae Kim ),문수현 ( Su Hyun Mun ),서윤수 ( Yun Soo Seo ),강다혜 ( Da Hye Kang ),임재수 ( Jae Soo Lim ),김마룡 ( Ma Ryong Kim ),곽남원 ( Nam Won Kwak ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6
Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-кB). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and NF-кB in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and 250 μg/ml). SHWDS (250 μg/ml) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor NF-кB in nuclear, degradation of IkB α in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and NF-кB activation, thereby decreasing the expression of pro-inflammatory genes.