• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.63-71
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• 2000

An immunogenic, high molecular weight lipopolysaccharide (LPS)-protein complex isolated from a potassium thioncyanate extract of a Pasteurella multocida (P multocida ; strain P-2383, capsular type A and somatic type 3) was characterized. Chemical analysis of the complex by gas chromatography on a capillary column demonstrated that this complex contained most of the chemical constituents characteristic of LPS extracted by the phenol-water methed from the whole bacterium. However, there was proportionately more carbohydrate than fatty acid in the complex in contrast to LPS in which fatty acid seemed to be in excess. When toxicity of the complex was evaluated in 10-day-old chicken embryos, the complex was less toxic ($LD_{50}=12.72{\mu}g$) than the purified LPS ($LD_{50}=0.44{\mu}g$). The $LD_{50}$, of the LPS moiety extracted from the complex was $5.24{\mu}g$. Composition of the complex was analyzed by SDS-PAGE with silver staining and Western immunoblotting. The complex did not migrate through the polyacrylamide gel unless dissociated with SDS. The complex dissociated with SDS contained at least 32 different protein and polysaccharide components: 18 components reacted with an antiserum against the complex. There was no significant compositional variation between the complexes from different strains, but quantitative differences in individual components were noted. When cross-protectivity of the complex was evaluated in mice, this complex provided substantial protection not only against the homologous bacteriun but also against different P multocida strains of the same serotype. LPS-protein complexes isolated by the same method from other strains also induced protection against an challenge with P-2383.

• Journal of Microbiology
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• v.39 no.4
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• pp.314-320
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• 2001

Macrophages stimulated by lipopolysaccharide(LPS) from gram negative bacteria undergo activation of a group of immediate early genes including Rantes. The mouse Rantes gene promoter region contains an LPS rsponsive element(LPE) We detected 3 specific bands termed B1, B2 and 3 formed by the interaction of the LPE and proteins found in LPS-stimulated RAW 367.7 cells. An additional band B4 was determined to be an Ap-1 binding protein. The B1 band appears within 1 hour of LPS nuclear extracts from LPS-stimulation, and this protein kinase enhances B1 and formation. The B1 band can be converted to band B2/B3 by adding specific heparin column fraction purified Ser/Thr specific protein phosphatases PP-1 and PP-2A can stimulate the same conversion to about the same extent. Thus, the formation of the LRE sequence binding complex appears to be regulated by Ser/Thr protein kinase and one or more Ser/Thr specific phosphatases. At least four proteins are involved in the trgulation of the LRE-dependent Rants experssion: two binding factors that bind directly to the target sequences. and two factors that control their binding. The future purification and characterization of these binding pro-teins will reveal in detail the mechanism of Rantes gene activation after LPS stimulation.

• The Korea Journal of Herbology
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• v.38 no.1
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• pp.11-19
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• 2023

Objectives : Neuroinflammation is a common pathological mechanism of neurodegenerative diseases, and the development of therapeutic agents is urgently needed. Red ginseng has been known to be good for the immune stimulation in Eastern Asia. Although the immuno-stimulatory activity of red ginseng are already known, the neuro-protective effects of cultivated red ginseng with fermented complex mushroom-cereal mycelium (RGFM) have not been conducted. Thus, in this study, we tried to investigate the anti-neuroinflammatory effect of RGFM water extract on lipopolysaccharide (LPS) stimulated BV2 cells. Methods : BV2 cells were pretreated with RGFM 1 h prior to LPS exposure. To determine the neuro-protective effects of RGFM water extract, we measured the expression of inflammatory mediators including inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and nitric oxide (NO) and pro-inflammatory cytokines such as interleukin (IL)-1𝛽, IL-6 and tumor necrosis factor (TNF)-𝛼 in LPS-stimulated BV2 cells. In addition, to find out the regulatory mechanism of RGFM water extract, we assessed the protein levels of mitogen-activated protein kinases (MAPKs) and inhibitory 𝜅B𝛼 (I𝜅B𝛼) by western blotting. Results : In our study, treatment of RGFM reduced the mRNA expression of iNOS and COX-2 and suppressed NO production in LPS-stimulated BV2 cells. Additionally, the secretion of IL-1𝛽 and TNF-𝛼 but not IL-6 was significantly inhibited by RGFM. Furthermore, RGFM water extract inhibited the phosphorylation of c-Jun N-terminal kinase (JNK). Conclusions : Taken together, these findings suggest that RGFM water extract has a protective effect on neuroinflammation through inhibition of JNK.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.878-883
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• 2008

Conchiolin is a complex protein which is secreted by the mollusc's outer epithelium to be the organic basis of mollusc shell. This study is to investigate a potential anti-inflammatory activity of conchiolin of oyster shell (COS). We tested the effects of COS on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and prostaglandin E2 (PGE 2) in a murine macrophage cell line, RAW 264.7. COS inhibited production of NO and PGE2 in a dose dependent manner and also decreased the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6). These results suggest that COS can inhibit production of pro-inflammatory mediators and might be a useful source to treat inflammation.

• Biomedical Science Letters
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• v.18 no.4
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• pp.369-376
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• 2012

Platycodon grandiflorum is a medicinal herb that is used to treat pulmonary and respiratory allergic disorders. The objective of this study was to investigate the protective effects of ethyl acetate extract of Platycodon grandiflorum (PGEA) against inflammation and to discern the molecular mechanism of PGEA in lipopolysaccharide (LPS)-induced signal pathways in RAW264.7 macrophage cells. PGEA suppressed the generation of nitric oxide (NO) and the expression of inducible NO synthase induced by LPS in RAW264.7 cells, and inhibited the release of pro-inflammatory cytokines induced by LPS in RAW264.7 cells. Western blot analysis showed that PGEA suppressed LPS-induced phosphorylation of p38 and c-Jun N-terminal kinase (JNK) but not extracellular signal-regulated kinase and $I{\kappa}-B{\alpha}$ degradation. Inactivation of JNK and p38 was effectively alleviated by PGEA, which subsequently affected the activation of c-Jun and c-Fos, which are the essential components of the activator protein-1 (AP-1) transcription complex. Taken together, the results indicate PGEA suppress the activation of p38, JNK, and AP-1, thereby inhibiting the generation of NO and pro-inflammatory cytokines, which affect the regulation of inflammation. PGEA may be useful for the treatment of various inflammatory diseases.

• The Journal of the Convergence on Culture Technology
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• v.9 no.1
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• pp.717-724
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• 2023

we are investigated the anti-inflammatory effects of Safflower and Mother wort Ethanol Complex Extracts(SEC) on lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results demonstrated that pretreatment of SEC(500㎍/mL) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects by Safflower and Mother wort Ethanol Complex Extracts were observed in the following. Safflower and Mother wort Ethanol Complex Extracts 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 Safflower and Mother wort Ethanol Complex Extracts exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of Safflower and Mother wort Ethanol Complex Extracts. Therefore, Safflower and Mother wort Ethanol Complex Extracts could be regarded as a potential source of natural anti-inflammatory agents.

• Journal of Applied Biological Chemistry
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• v.59 no.1
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• pp.37-43
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• 2016

Bioactive components of ultra-fine ground Saururus, the extraction yield increases when the leaves are ultra-fine ground. Comparison of normal-ground and ultra-fine ground Saururus chinensis leaves showed that the solid content and antiinflammatory activity of ultra-fine ground extracts was higher than that of normal-ground extracts. Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells were treated with different concentrations of Saururus chinensis extract and the amount of nitric oxide (NO) was determined; LPS-treated cells produced 2 times more NO than cells that were not treated with LPS. Moreover, the NO production in cells treated with Saururus chinensis extract was inhibited in a concentration-dependent manner. Because the stimulant-induced NO production is regulated by the inducible nitric oxide synthase (iNOS), we measured the iNOS protein level to elucidate the mechanism by which the NO production was inhibited. We found that the amount of iNOS decreased dose-dependently. It was reduced by 53% at a Saururus chinensis extract concentration of $100{\mu}g/mL$. The protein expression of cyclooxygenase-2 (COX-2) in LPS-treated Raw 264.7 cells was inhibited by 31% at $100{\mu}g/mL$ of Saururus chinensis extract. Gel shift of the nuclear factor kappa B-DNA complex occurred in LPS-treated cells and the intensity of the band decreased gradually in a concentration-dependent manner. Ultra-fine ground Saururus chinensis extract had a concentration-dependent inhibitory effect on the production of prostaglandin $E_2$, tumor necrosis factor ${\alpha}$, interleukin $1{\beta}$ (IL-$1{\beta}$), IL-6, and IL-8 in LPS-treated Raw 264.7 cells, i.e., at $50{\mu}g/mL$ of Saururus chinensis extract, their levels were decreased by 53, 67, 52, 37, and 21% respectively.

• Journal of Acupuncture Research
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• v.23 no.2
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• pp.33-46
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• 2006

Alzheimer's disease (AD) is the most prevalent form of neurodegenerative disease associated with aging in the human population. This disease is characterized by the extracellular deposition of beta-amyloid peptide $(A{\beta})$ in cerebral plaques. $A{\beta}$ is derived from the ${\beta}-amyloid$ precursor protein (APP) by the enzymes, ${\beta}-$ and ${\eta}o-secretase$. Compounds that ${\beta}-$ or ${\eta}o-secretase$ inhibit activity, can reduce the production of $A{\beta}$ peptides, and thus have therapeutic potential in the treatment of AD. Increasing body of evidence has been demonstrated that Bee Venom(BV) Acupuncture could compete with complex protein involving in multiple step of $NF-{\kappa}B$ activation and exert the anti-inflammatory potential of combined inhibition of the prostanoid and nitric oxide synthesis systems by inhibition of IKK and $NF-{\kappa}B$. In this study, I investigated possible effects of BV on memory dysfunction caused by lipopolysaccharide (LPS) and $A{\beta}$ through inhibition of secretases activities and $A{\beta}$ aggregation. I examined the improving effect of BV on the LPS (2.5 mg/Kg, i.p.)-induced memory dysfunction using passive avoidance response and water maze tests in the mice. BV (0.84, $1.67\;{\mu}g/ml$) reversed the LPS-induced memorial dysfunction in dose dependent manner. BV also dose-dependently attenuated LPS-induced ${\beta}$ and ${\eta}o-secretase$ activities in cerebral cortex and hippocampus of the mice brain. This study therefore suggests that BV acupuncture method may be useful for prevention of development or progression of AD.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.87-93
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• 2013

Objectives : Degenerative arthritis arises from several physiological factors. The purpose of this study is to investigate the beneficial effects of Phyto-extract Complex (CME) on degenerative arthritis. Methods : CME is composed of extracts of mulberry (Morus alba L.) fruit, mulberry leaves and black beans (Glycine max (L.) Merr.). To measure the toxicity of CME, we performed the single-dose toxicity study. For the evaluation of its effects on degenerative arthritis, we examined the inhibition of cyclooxygenase-2 (COX-2) activity, using in vitro enzyme activity assay, the reduction of protein expression of COX-2, 5-lipoxygenase (5-LO), and inducible nitric oxide synthase (iNOS) in RAW264.7 cells which were stimulated by lipopolysaccharide (LPS). We also examined the serum level of prostaglandins (PGs) and injury of the knee joint cartilage, using animal model of degenerative arthritis induced by mono-sodium iodoacetate (MIA). Results : CME did not have any toxicity in single-dose toxicity study. The CME inhibited the activity of COX-2 and could reduce the protein expression of COX-2, 5-LO and iNOS in RAW264.7 cells. The CME also reduced the serum level of PGs and prevented from the cartilage injury of knee joint in animal model of degenerative arthritis induced by MIA. Conclusions : Taken altogether, the CME could be useful for the improvement of degenerative arthritis through its various anti-inflammatory activities and prevention from the cartilage injury of knee joint.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.195-204
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• 2021

Cereblon (CRBN), a substrate receptor of cullin 4-RING E3 ligase (CRL4) regulates the ubiquitination and degradation of c-Jun, mediating the lipopolysaccharide-induced cellular response. However, the upstream signaling pathway that regulates this process is unknown. In this study, we describe how endoplasmic reticulum (ER) stress reversely regulates sequestosome-1 (p62)and c-Jun protein levels. Furthermore, our study reveals that expression of p62 attenuates c-Jun protein levels through the ubiquitinproteasome system. Conversely, siRNA knockdown of p62 elevates c-Jun protein levels. Immunoprecipitation and immunoblotting experiments demonstrate that p62 interacts with c-Jun and CRBN to form a ternary protein complex. Moreover, we find that CRBN knockdown completely abolishes the inhibitory effect of p62 on c-Jun. Using brefeldin A as an inducer of ER stress, we demonstrate that the p62/c-Jun axis participates in the regulation of ER stress-induced apoptosis, and that CRBN is required for this regulation. In summary, we have identified an upstream signaling pathway, which regulates p62-mediated c-Jun degradation. Our findings elucidate the underlying molecular mechanism by which p62/c-Jun axis regulates the ER stress-induced apoptosis, and provide a new molecular connection between ER stress and apoptosis.