• The Korea Journal of Herbology
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• v.37 no.5
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• pp.83-88
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• 2022

Objectives : Jakyakgamcho-tang (JGT) has been traditionally used to treat muscular convulsion and pain in South Korea. According to recent studies, JGT has been reported to have anti-depression, anti-inflammation, anti-oxidative, anti-diabetics, anti-spasm and analgesic effects, but studies on its anti-neuroinflammatory and neuroprotective effect have not been deeply conducted. Thus, we investigated the anti-neuroinflammatory activity of JGT on lipopolysaccharide (LPS)-stimulated mouse microglia cells. Methods : To investigate the anti-neuroinflammatory effects of JGT on BV2 microglial cells, we examined the production of nitric oxide (NO) using griess assay, and mRNA expressions of pro-inflammatory cytokines such as interleukin (IL)-1𝛽, IL-6, and tumor necrosis factor (TNF)-𝛼 using real time RT-PCR. Furthermore, to determine the regulating mechanisms of JGT, we investigated the heme oxygenase (HO)-1 by real time RT-PCR. Results : Pre-treatment of JGT effectively decreased NO production in LPS-stimulated BV2 cells at concentrations without cytotoxicity. Additionally, JGT significantly suppressed the production of IL-1𝛽, IL-6, and TNF-𝛼 in LPS-stimulated BV2 cells. Furthermore, JGT activated the HO-1 expression, which is one of the immunomodulatory signaling molecules. And the abolishment of HO-1 by tin protoporphyrin IX (SnPP, the HO-1 inhibitor) reversed the anti- inflammatory activity of JGT in LPS-stimulated BV2 cells. Conclusions : Our results suggest that the JGT has anti-neuroinflammatory effect through the activation of HO-1 in LPS-stimulated BV2 cells. Thereby, JGT could expected to be used for the prevention and treatment of neurodegenerative disease related to neuroinflammation.

• The Journal of Korean Medicine
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• v.43 no.4
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• pp.20-32
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• 2022

Objectives: Kyungok-go (KOG) is a traditional multi-herbal medicine commonly used for enforcing weakened immunity for long time. Recently, there are several reports that KOG has anti-inflammatory and immuno-stimulatory activities in many experimental models. However, the protective effects of KOG on neuronal inflammation are still undiscovered. Thus, we investigated the neuro-protective activity of KOG on lipopolysaccharide (LPS)-stimulated mouse microglia cells. To find out KOG's anti-neuroinflammatory effects on microglial cells, we examined the production of nitrite using griess assay, and mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α using real time RT-PCR. In addition, to examine the regulating mechanisms of KOG, we investigated the protein expression of mitogen-activated protein kinases (MAPKs) and Iκ-Bα by western blot. KOG inhibited the elevation of nitrite, iNOS and COX-2 on LPS-stimulated BV2 cells. Also, KOG significantly inhibited the pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α on LPS-stimulated BV2 microglial cells. Moreover, KOG inhibited the activation of c-Jun N-terminal kinase (JNK), P38 and degradation of Iκ-Bα but not the activation of extracellular signal regulated kinase (ERK) on LPS-stimulated BV2 microglial cells. These results showed KOG has the anti-inflammatory effects through the inhibition on nitrite, iNOS, COX-2, IL-1β, IL-6, and TNF-α via the deactivation of JNK, p38 and nuclear factor (NF)-κB on LPS-stimulated BV2 microglial cells. Thereby, KOG could offer the new and promising treatment for neurodegenerative disease related to neuroinflammation.

• The Korea Journal of Herbology
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• v.38 no.6
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• pp.45-52
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• 2023

Objectives : Effects of baicalein (BA) on oxidative stress in RAW 264.7 mouse macrophages stimulated with peptidoglycan (PG) and lipopolysaccharide (LPS) were investigated. Methods : RAW 264.7 co-stimulated with LPS and PG were incubated with BA at concentrations of 25 and 50 µM. Incubation time was 18 h, 20 h, 22 h, 24 h, and 26 h. After incubation, the production of hydrogen peroxide in RAW 264.7 was measured with dihydrorhodamine 123 assay. Additionally, RAW 264.7 stimulated with PG were incubated with BA at concentrations of 25 and 50 µM for 24 h. After incubation, NO production was evaluated by griess reagent assay. Results : BA significantly inhibited hydrogen peroxide productions (p <0.05). In details, production of hydrogen peroxide in 'LPS and PG'-stimulated RAW 264.7 treated for 18 h with BA at concentrations of 25 and 50 µM was 91.27% and 89.22% of the control group treated with LPS and PG only, respectively; the production of hydrogen peroxide for 20 h was 92.19% and 90.58%, respectively; production of hydrogen peroxide for 22 h was 91.69% and 89.89%, respectively; production of hydrogen peroxide for 24 h was 92.4% and 90.19%, respectively; production of hydrogen peroxide for 26 h was 91.7% and 89.04%, respectively. Additionally, BA at the concentration of 50 and 100 µM significantly inhibited NO production in PG-induced RAW 264.7 (p <0.05). Conclusions : BA might have anti-oxidative activity related to its inhibition of hydrogen peroxide production in 'LPS and PG'-stimulated RAW 264.7 macrophages.

• Toxicological Research
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• v.13 no.1_2
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• pp.111-116
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• 1997

In the present study, we investigated the neutralization activity of various antimicrobials against lipopolysaccharide (LPS) as well as interaction between two antimicrobials (enrofioxacin and coilstin) using checkerboard method. The neutralization activity of antimicrobials used in the test was assayed by means of LAL chromogenic test after reaction of LPS with colistin, enorfioxacin, ampicillin, polymyxin B, oxytetracycline, streptomycin, and erythromycin. As the results, the neutralization activity of coltstin and polymixin B had a more stronger than that of tested other antimicrobials. In bacterial culture broth, the best neutralization activity of the antibiotics was also shown to coltstin and polymixin B. Meanwhile, It was shown to have synergism between enorfloxacin and coltstin on the basts of FIC (fractional inhibition concentration). The FIC of enorfioxacin-colistin combinations was 0.50-1.03 to Staphylococcus aureus R-209, 1.03-1.06 to Salmonella typhimurium, 0.75-1.25 to Bordetella Bronchtseptica and 1.02-1.25 to E. coli K88ab. On the basts of the above results, the present study may be of clinical usefulness in the choice of an antibiotic therapy for severe sepsts in animals.

• International Journal of Internet, Broadcasting and Communication
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• v.12 no.3
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• pp.139-148
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• 2020

Red rose petals are usually disposed but they are an abundant source of phenolics and traditionally used as food supplement and as herbal medicine. Of the Various phenolics, they are known to have anticancer, antioxidant, and anti-inflammatory properties. In this study, we investigated the anti-inflammatory effects of red rose ethanolic extracts (GRP) on lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results demonstrated that pretreatment of GRP (500㎍/mL) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects by red rose petals were observed in the following. Red rose petals inhibited the translocation of NF-κB from the cytosol to the nucleus via the suppression of IκB-α phosphorylation and also inhibited LPS-stimulated NF-κB transcriptional activity. These findings suggest that red rose petals exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of red rose petals. Therefore, red rose petals could be regarded as a potential source of natural anti-inflammatory agents.

• BMB Reports
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• v.36 no.5
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• pp.520-523
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• 2003

The proliferation and differentiation signals of myelogeneous U937 cells are provided by extracellular stimuli, such as lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA). In a DNA-native-polyacrylamide gel assay system, we demonstrated that a particular nuclease activity is expressed in PMA-stimulated U937 cells and secreted into the culture medium. The nuclease activity was induced in U937 cells by LPS treatment, while the secretion of the enzyme was undetected in the culture medium. Therefore, it is likely that the expression and secretion of the particular nuclease in U937 cells are controlled by extracellular stimulations, such as PMA and LPS treatment.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1423-1426
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• 2008

Nuruk, which is a natural inoculator and source of amylolytic enzymes, is used in Korean traditional rice wine. A methanol extract of nuruk (NE) attenuated lipopolysaccharide (LPS)-induced nitrite and interleukin (IL)-6 in RAW 264.7 cells. Both the n-hexane and water fractions from NE (MEH and MW, respectively) inhibited the production of nitrite and IL-6 in RAW 264.7 cells. MEH and MW also inhibited the LPS-induced inducible nitric oxide synthase expression. Further, and MEH protected against the LPS-induced activation of p38 mitogen-activated protein kinase. Together, these results indicate that nuruk may contribute to the anti-inflammatory and cancer-preventive effects of Korean traditional rice wine.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.10a
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• pp.165-165
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• 2001

The effects of betaine or taurine on hepatotoxicity induced by lipopolysaccharide (LPS) were examined in adult male SD rats. Rats were provided with drinking water containing either 1 % betaine or taurine for 2 weeks prior to challenge with LPS (5 mg/kg, iv). Supplement of betaine or taurine protected the animals from induction of LPS hepatotoxicity as measured by changes in aspartate aminotrassferase (AST), alanine aminotransferase (ALT) activities and total bilirubin levels in serum, and hepatic glutathione contents.(omitted)

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.111-121
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• 2005

Human placental extract (HPE), which is prepared from the placenta of healthy pregnant females, has been widely used in clinical field. HPE is known to possess anti-inflammatory, anti-viral, anti-oxidative, anti-mutagenic, and analgesic properties. In this study, the effect of HPE against lipopolysaccharide (LPS)-induced inflammation was investigated. From the present results, HPE was shown to suppress prostaglandin E2 synthesis (PGE2) and nitric oxide (NO) production by inhibition on the LPS-stimulated enhancement of the cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expressions in mouse BV2 microglial cells. These results suggest that HPE may offer a valuable mean of therapy for the treatment of brain inflammatory diseases by attenuating LPS-induced PGE2 and NO production.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.1
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• pp.28-35
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• 2008

Purpose: The nitric oxide (NO) release by inducible nitric oxide synthase (iNOS) is the key events in macrophage response to lipopolysaccharide (LPS) which is suggested to be a crucial mediator for inflammatory and innate immune responses. NO is an important mediator involved in many host defense action and may also lead to a harmful host response to bacterial infection. However, given the importance of iNOS in a variety of pathophysiological conditions, control of its expression and signaling events in response to LPS has been the subject of considerable investigation. Materials and Methods: The Raw264.7 macrophage cell line was used to observe LPS-stimulated iNOS expression. The expression of iNOS is observed by Western blot analysis and real-time RT-PCR. Protein kinase C $(PKC)-{\alpha}$ overexpressing Raw264.7 cells are established to determine the involvement of $PKC-{\alpha}$ in LPS-mediated iNOS expression. $NF-{\kappa}B$ activity is measured by $I{\kappa}B{\alpha}$ degradation and $NF-{\kappa}B$ luciferase activity assay. Results: We found that various PKC isozymes regulate LPS-induced iNOS expression at the transcriptional and translational levels. The involvement of $PKC-{\alpha}$ in LPS-mediated iNOS induction was further confirmed by increased iNOS expression in $PKC-{\alpha}$ overexpressing cells. $NF-{\kappa}B$ dependent transactivation by LPS was observed and $PKC-{\alpha}$ specific inhibitory peptide abolished this activation, indicating that $NF-{\kappa}B$ activation is dependent on $PKC-{\alpha}$. Conclusion: Our data suggests that $PKC-{\alpha}$ is involved in LPS-mediated iNOS expression and that its downstream target is $NF-{\kappa}B$. Although $PKC-{\alpha}$ is a crucial mediator in the iNOS regulation, other PKC isozymes may contribute LPS-stimulated iNOS expression. This finding is needed to be elucidated in further study.