• Journal of Applied Biological Chemistry
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• 제52권1호
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• pp.21-27
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• 2009

인삼은 가공처리방법에 따라 홍삼과 발효홍삼으로 구분된다. 또한, 이들의 가공처리방법에 따라 인삼의 효능을 나타내는 사포닌의 함량이 차이가 있다. 따라서, 본 연구에서 인삼, 홍삼 및 발효홍삼이 대식세포에서 LPS에 의한 염증에 대한 항염효과 및 그 기전을 규명하고자 하였다. 마우스의 대식세포인 RAW264.7 세포에서 LPS에 의해 유도되는 염증관련인자인 NO 및 COX-2의 발현 및 TNF-$\alpha$, INF-$\gamma$ 그리고 NF-${\kappa}B$의 활성을 인삼, 홍삼 그리고 발효홍삼에 의한 항염효과 차이를 비교 하였다. 그 결과, 인삼 및 홍삼 그리고 발효홍삼 모두에서 LPS에 의한 NO의 생성을 억제시키는 것을 확인하였으며, TNF-$\alpha$ 및 INF-$\gamma$의 생성 또한 억제시키는 것을 알 수 있었다. 또한, 인삼 및 홍삼 그리고 발효홍삼 모두 COX-2의 발현 및 LPS에 의한 $I{\kappa}B$의 인산화를 억제시킴으로써 NF-${\kappa}B$의 활성을 억제시키는 것임을 알 수 있었다. 홍삼이 인삼과 발효홍삼에 비하여 NO의 생성을 더 효과적으로 억제시키는 것은 각각의 제조과정에서 나타나는 인삼사포닌의 조성의 차이에 따른 것으로 추정된다. 본 연구는 단순한 동물세포 수준에서의 비교 차이이며 좀 더 정확한 기전의 규명을 위해서는 향후 동물실험을 통한 비교 실험이 수행되어야 할 것이다.

• Journal of Microbiology and Biotechnology
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• 제26권3호
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• pp.469-476
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• 2016

This study was undertaken to evaluate the immunomodulatory effect of dead nano-sized Lactobacillus plantarum (nLp) in RAW 264.7 cells and murine primary splenocytes. nLp is a dead, shrunken, processed form of L. plantarum nF1 isolated from kimchi (a traditional Korean fermented cabbage) and is less than 1 μm in size. It was found that nLp treatment stimulated nitric oxide (NO) production more in RAW 264.7 macrophages than pure live L. plantarum (pLp), and that the stimulatory properties were probably largely derived from its cell wall. In addition, nLp induced murine splenocyte proliferation more so than pLp; in particular, a high dose of nLp (1.0 × 10¹¹ CFU/ml) stimulated proliferation as much as lipopolysaccharide at 2 μg/ml. Moreover, according to our cytokine profile results in splenocytes, nLp treatment promoted Th1 (TNF-α, IL-12 p70) responses rather than Th2 (IL-4, IL-5) responses and also increased Th17 (IL-6, IL-17A) responses. Thus, nLp stimulated NO release in RAW 264.7 cells and induced splenocyte proliferation more so than pLp and stimulated Th1 and Th17 cytokine production. These findings suggested that dead nLp has potential use as a functional food ingredient to improve the immune response, and especially as a means of inducing Th1/Th17 immune responses.

• 생약학회지
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• 제47권2호
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• pp.150-157
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• 2016

Earth worm (Eisenia andrei) and clam worm (Perinereis linea) have been used as anti-bacterial and anti-inflammatory agent. However, it is unclear how they exerted their physiological effects in macrophages. In this experiment, the anti-inflammatory and antioxidant effects of clam worm extract (CWE) and earth worm extract (EWE) in RAW264.7 cells were examined by measuring MDA, catalase, SOD, GSH-Px and inflammatory cytokines (nitric oxide, iNOS, interleukin-$1{\beta}$ and tumor necrosis factor-${\alpha}$). Treatment with CWE significantly increased the activities of catalase, SOD and GSH-Px in RAW264.7 cells and decreased the level of MDA. Interestingly, treatment with CWE induced more activities of SOD than EWE. In addition, CWE decreased NO production, iNOS, COX-2, TNF-${\alpha}$ and IL-$1{\beta}$ in RAW264.7. The EWE also decreased NO production and iNOS, but increased COX-2 and IL-$1{\beta}$ suggesting that CWE could be better resources for anti-inflammatory and antioxidant agent than EWE. Taken together, these results indicate that CWE has the potential as a natural antioxidant and a therapeutic for inflammation-related diseases.

• 한국자원식물학회지
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• 제31권6호
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• pp.612-621
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• 2018

Honey used as conventional medicine has various pharmacological properties. In the honey and anti-inflammatory effect, Gelam honey and Manuka honey has been reported to exert anti-inflammatory activity. However, the anti-inflammatory effect and potential mechanisms of acacia honey (AH) are not well understood. In this study, we investigated anti-inflammatory activity and mechanism of action of AH in LPS-stimulated RAW264.7 cells. AH attenuated NO production through inhibition of iNOS expression in LPS-stimulated RAW264.7 cells. AH also decreased the expressions of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ as pro-inflammatory cytokines, and MCP-1 expression as a pro-inflammatory chemokine. In the elucidation of the molecular mechanisms, AH decreased LPS-mediated $I{\kappa}B$-${\alpha}$ degradation and subsequent nuclear accumulation of p65, which resulted in the inhibition of $NF-{\kappa}B$ activation in RAW264.7 cells. AH dose-dependently suppressed LPS-mediated phosphorylation of ERK1/2 and p38 in RAW264.7 cells. In addition, AH significantly inhibited ATF2 phosphorylation and nuclear accumulation of ATF2 in LPS-stimulated RAW264.7 cells. These results suggest that AH has an anti-inflammatory effect, inhibiting the production of pro-inflammatory mediators such as NO, iNOS, $TNF-{\alpha}$, IL-6, $IL-1{\beta}$ and MCP-1 via interruption of the $NF-{\kappa}B$ and MAPK/ATF2 signaling pathways.

• 동의생리병리학회지
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• 제19권5호
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• pp.1225-1232
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• 2005

Based on traditional medicine theories, GC, an extract from 5 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present studies was done to investigate whether GC has inhibitory effects on activation of fibroblast-like sinoviocytes isolated from a RA patient. In tumor necrosis factor-${\alpha}(TNF-{\alpha}$)/ interleukin-IL-$1{\beta}$(IL-$1{\beta}$) treated human sinoviocytes, the mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GC($10{\mu}g/ml$) significantly suppressed the expression of proinflammatory cytokines and chemokines such as $TNF-{\alpha}$, IL-6 and IL-8 compared with the control, but not $IL-1{\beta}$, The mRNA level of intracellular adhesion molecule-1 (ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GC. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). Furthermore, GC dramatically raised the mRNA levels of tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), while those of matrix metalloproteinase-3 were significantly lowered. Taken together, these data suggested that GC might suppress the activation of sinoviocytes in hRA.

• 대한한의학방제학회지
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• 제25권4호
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• pp.537-551
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• 2017

Objectives : Jinnoe-san (JNS) is a novel herbal formula consisting of five oriental medicinal herbs including Polygalae Radix, Prunellae Spica, Perillae Herba, Betulae Cortex, and Lonicerae Flos. In this study, we investigated the effects and molecular mechanism of JNS on Parkinson's disease in vitro model. Methods : The effects of JNS on 1-methyl-4-phenylpyridinium ($MPP^+$)-induced cell death in SH-SY5Y cells were evaluated with a cell viability assay, flow cytometry, and western blots analysis. The effects of JNS on lipopolysaccharide (LPS)-stimulated BV2 microglia were determined with a nitric oxide (NO) assay, enzyme linked immunosorbent assays, and western blots analysis. Result : $MPP^+$-induced cell death in SH-SY5Y cells was significantly reduced by JNS pre-treatment in a dose-dependent manner. JNS inhibited the production of reactive oxygen species, mitochondria dysfunction, and apoptosis induced by $MPP^+$ in SH-SY5Y cells. Furthermore, JNS significantly activated Akt and ERK in SH-SY5Y cells and the ability of JNS to prevent mitochondria dysfunction by $MPP^+$ was antagonized by pre-treatment of LY294002 and PD98059, an Akt and ERK inhibitor, respectively. In addition, JNS inhibited LPS-induced NO and $PGE_2$ production as well as iNOS expression and secretion of TNF-${\alpha}$, pro-inflammatory cytokines without affecting the cell viability. JNS also suppressed LPS-induced ERK activation. Conclusions : These results demonstrate that JNS has a protective effect on the dopaminergic neurons against $MPP^+$-induced neurotoxicity and anti-inflammatory effect on the LPS-stimulated microglia. These findings provide evidences for JNS to be considered as a new prescription for treating Parkinson's disease.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.97-97
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• 2018

Honey used as conventional medicine has various pharmacological properties. In the honey and anti-inflammatory effect, Gelam honey and Manuka honey has been reported to exert anti-inflammatory activity. However, the anti-inflammatory effect and potential mechanisms of acacia honey (AH) are not well understood. In this study, we investigated anti-inflammatory activity and mechanism of action of AH in LPS-stimulated RAW264.7 cells. AH attenuated NO production through inhibition of iNOS expression in LPS-stimulated RAW264.7 cells. AH also decreased the expressions of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ as pro-inflammatory cytokines, and MCP-1 expression as a pro-inflammatory chemokine. In the elucidation of the molecular mechanisms, AH decreased LPS-mediated $I{\kappa}B-{\alpha}$ degradation and subsequent nuclear accumulation of p65, which resulted in the inhibition of $NF-{\kappa}B$ activation in RAW264.7 cells. AH dose-dependently suppressed LPS-mediated phosphorylation of ERK1/2 and p38 in RAW264.7 cells. In addition, AH significantly inhibited ATF2 phosphorylation and nuclear accumulation of ATF2 in LPS-stimulated RAW264.7 cells. These results suggest that AH has an anti-inflammatory effect, inhibiting the production of pro-inflammatory mediators such as NO, iNOS, $TNF-{\alpha}$, IL-6, $IL-1{\beta}$ and MCP-1 via interruption of the $NF-{\kappa}B$ and MAPK/ATF2 signaling pathways.

• 대한본초학회지
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• 제31권2호
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• pp.63-69
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• 2016

Objectives : The aerial parts of Agrimonia pilosa Ledeb (Agrimoniae Herba; AH) has been traditionally used as a Korean medicine to treatment of abdominal pain, sore throat, headaches, bloody discharge, parasitic infections and eczema. In this study, we investigated the effect of AH ethanol extract on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophage cells.Methods : AH was extracted by 30% ethanol (AH-E). Raw264.7 cells were treated with AH-E extract at different concentrations for 30 min and then stimulated with LPS (1㎍/㎖) or without for indicated times. Cell viability was measured by MTT assay, and nitric oxide (NO) production was measured by Griess assay. The expression of inflammatory mediators, iNOS and COX-2 and inflammatory cytokines, TNF-α, IL-1β, and IL-6 was detected by RT-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was analyzed by Western blot. Also, the expression of NF-κB in nuclear and cytosol was detected by Western blot.Results : AH-E extract significantly decreased LPS-induced NO production in RAW264.7 cells. AH-E extract inhibited the mRNA expression of iNOS, COX-2, TNF-α, IL-1β, and IL-6 in LPS-stimulated cells with a dose-dependent manner. In addition, the phosphorylation of ERK, p38 and JNK MAPKs was also inhibited by AH-E extract. AP-E extract inhibited the nuclear translocation of NF-κB in LPS-stimulated cells.Conclusions : Our results suggest that AH-E extract has an anti-inflammatory activity in macrophages-mediated inflammation.

• Journal of Microbiology and Biotechnology
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• 제30권3호
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• pp.333-340
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• 2020

Macrophages are the cells of the first-line defense system, which protect the body from foreign invaders such as bacteria. However, Gram-negative bacteria have always been the major challenge for macrophages due to the presence of lipopolysaccharides on their outer cell membrane. In the present study, we evaluated the effect of phloretin, a flavonoid commonly found in apple, on the protection of macrophages from Escherichia coli infection. RAW 264.7 cells infected with standard E. coli, or virulent E. coli K1 strain were treated with phloretin in a dose-dependent manner to examine its efficacy in protection of macrophages. Our results revealed that phloretin treatment reduced the production of nitric oxide (NO) and generation of reactive oxygen species along with reducing the secretion of proinflammatory cytokines induced by the E. coli and E. coli K1 strains in a concentration-dependent manner. Additionally, treatment of phloretin downregulated the expression of E. coli-induced major inflammatory markers i.e. cyclooxygenase-2 (COX-2) and hemeoxygenase-1 (HO-1), in a concentration dependent manner. Moreover, the TLR4-mediated NF-κB pathway was activated in E. coli-infected macrophages but was potentially downregulated by phloretin at the transcriptional and translational levels. Collectively, our data suggest that phloretin treatment protects macrophages from infection of virulent E. coli K1 strain by downregulating the TLR4-mediated signaling pathway and inhibiting NO and cytokine production, eventually protecting macrophages from E. coli-induced inflammation.

• 대한본초학회지
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• 제29권4호
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• pp.13-20
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• 2014

Objectives : Atotang was composed of 10 kinds of traditional medicinal herb. This research was performed to examine biological effects of Atotang for the development of prescription on atopic dermatitis. Methods : Atotang was extracted with 80% EtOH. Free radical scavenging assay has tested for anti-oxidative activity as well as the contents of total polyphenol. We observed the production of ROS, nitric oxide(NO) and the inflammatory cytokines such as interleukin-1beta(IL-${\beta}$), IL-6, tumor necrosis factor-alpha(TNF-${\alpha}$), Prostaglandin E2($PGE_2$) in Raw 264.7 cells stimulated by LPS. We used Disc diffusion method to investigate antibacterial activity on Candida albicans, Staphylococcus aureus and Staphylococcus epidermis. Result : Content of total phenolic compound of Atotang was 36.3 mg/g ext. DPPH and ABTS scavenging activities were 77% and 46% at 200 ug/ml respectively, showing dose-dependent increase. The amounts of ROS and NO in RAW 264.7 cells were decreased by 30% and 19% at 200 ug/ml, respectively, showing dose-dependent decrease. The prodcution of IL-1beta, IL-6 and TNF-alpha in RAW 264.7 cells were decreased dose-dependently by 81%, 67%, and 20% at 200 ug/ml, respectively. Atotang was reduced LPS-stimulated production of $PGE_2$ by 33%. Atotang on C. albicans, S. aureus and S. epidermis was selected by a disc diffusion method and inhibition effect of the Atotang on the growth of S. epidermis was the greatest. Conclusion : The results indicated that Atotang showed biological activities showing anti-oxidant, anti-inflammatory and antibacterial effects. Based on these results, it is concluded that Atotang can be applied to the prescription on atopic dermatitis.