Anti-inflammatory activities of Scolopendra subspinipes mutilans in RAW 264.7 cells / 한국영양학회지

PURPOSE: The dried body of Scolopendra subspinipes mutilans has long been used as a traditional Korean medicinal food, but little is known about its mechanisms of action. In this study, we investigated the anti-inflammatory activities of Scolopendra subspinipes mutilans and possible mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. METHODS: Cytotoxicity of Scolopendra subspinipes mutilans extract (SSME) was measured by MTT assay, anti-inflammatory activities were analyzed by nitric oxide (NO) production, the expression of inducible NO synthase (iNOS) and the mRNA level of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-6 (IL-6). Nuclear translocation of nuclear factor-kappa B (NF-κB) p65 subunit and degradation of inhibitory kappa B (IκB) were examined by western blot. RESULTS: SSME inhibited LPS-induced NO production and iNOS expression without cytotoxicity. Up-regulation of LPS-induced pro-inflammatory cytokines, IL-1β and IL-6 was dose dependently attenuated by SSME. Exposure of pyrrolidine dithiocarbamate, an NF-κB specific inhibitor, accelerated the inhibitory effects of SSME on NO production and iNOS expression in LPS-stimulated cells. Moreover, translocation of NF-κB from the cytosol to the nucleus and degradation of IκB were decreased by treatment with SSME in LPS-induced cells. CONCLUSION: These results suggest that the SSME might have the inhibitory effects on inflammation, partly through inhibition of the NF-κB signaling pathway.

Inhibitory effect of Petalonia binghamiae on neuroinflammation in LPS-stimulated microglial cells / 한국영양학회지

PURPOSE: Neuroinflammation is mediated by activation of microglia implicated in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease. Inhibition of neuroinflammation may be an effective solution to treat these brain disorders. Petalonia binghamiae is known as a traditional food, based on multiple biological activities such as anti-oxidant and anti-obesity. In present study, the anti-neuroinflammatory potential of Petalonia binghamiae was investigated in LPS-stimulated BV2 microglial cells. METHODS: Cell viability was measured by MTT assay. Production of nitric oxide (NO) was examined using Griess reagent. Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was detected by Western blot analysis. Activation of nuclear factor κB (NF-κB) signaling was examined by nuclear translocation of NF-κB p65 subunit and phosphorylation of IκB. RESULTS: Extract of Petalonia binghamiae significantly inhibited LPS-stimulated NO production and iNOS/COX-2 protein expression in a dose-dependent manner without cytotoxicity. Pretreatment with Petalonia binghamiae suppressed LPS-induced NF-κB p65 nuclear translocation and phosphorylation of IκB. Co-treatment with Petalonia binghamiae and pyrrolidine duthiocarbamate (PDTC), an NF-κB inhibitor, reduced LPS-stimulated NO release compared to that in PB-treated or PDTC-treated cells. CONCLUSION: The present results indicate that extract of Petalonia binghamiae exerts anti-neuroinflammation activities, partly through inhibition of NF-κB signaling. These findings suggest that Petalonia binghamiae might have therapeutic potential in relation to neuroinflammation and neurodegenerative diseases.

Anti-apoptotic effect of fermented Citrus sunki peel extract on chemical hypoxia-induced neuronal injury / 한국영양학회지

PURPOSE: Neuronal apoptotic events induced by aging and hypoxic/ischemic conditions is an important risk factor in neurodegenerative diseases such as ischemia stroke and Alzheimer's disease. The peel of Citrus sunki Hort. ex Tanaka has long been used as a traditional medicine, based on multiple biological activities including anti-oxidant, anti-inflammation, and anti-obesity. In the current study, we examined the actions of fermented C. sunki peel extract against cobalt chloride (CoCl2)-mediated hypoxic death in human neuroblastoma SH-SY5Y cells. METHODS: Cell viability was measured by trypan blue exclusion. Expression of apoptosis related proteins and release of cytochrome c were detected by western blot. Production of intracellular reactive oxygen species (ROS) and apoptotic morphology were examined using 2',7'-dichlorofluorescin diacetate (DCF-DA) and 4',6-diamidino-2-phenylindole (DAPI) staining. RESULTS: Exposure to CoCl2, a well-known mimetic agent of hypoxic/ischemic condition, resulted in neuronal cell death via caspase-3 dependent pathway. Extract of fermented C. sunki peel significantly rescued the CoCl2-induced neuronal toxicity with the cell viability and appearance of apoptotic morphology. Cytoprotection with fermented C. sunki peel extract was associated with a decrease in activities of caspase-3 and cleavage of poly (ADP ribose) polymerase (PARP). In addition, increase in the intracellular ROS and release of cytochrome c from mitochondria to the cytosol were inhibited by treatment with extract of fermented C. sunki peel. CONCLUSION: Based on these data, fermented C. sunki peel extract might have a protective effect against CoCl2-induced neuronal injury partly through generation of ROS and effectors involved in mitochondrial mediated apoptosis.

Src Kinase Regulates Nitric Oxide-induced Dedifferentiation and Cyclooxygenase-2 Expression in Articular Chondrocytes via p38 Kinase-dependent Pathway

BACKGROUND: Nitric oxide (NO) in articular chondrocytes regulates dedifferentiation and inflammatory responses by modulating MAP kinases. In this study, we investigated whether the Src kinase in chondrocytes regulates NO-induced dedifferentiation and cyclooxygenase-2 (COX-2) expression. METHODS: Primary chondrocytes were treated with various concentrations of SNP for 24 h. The COX-2 and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin E(2) (PGE(2)) was determined by using a PGE(2) assay kit. Expression and distribution of p-Caveolin and COX-2 in rabbit articular chondrocytes and cartilage explants were determined by immunohistochemical staining and immunocytochemical staining, respectively. RESULTS: SNP treatment stimulated Src kinase activation in a dose-dependent manner in articular chondrocytes. The Src kinase inhibitors PP2 [4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo(3,4-d)pyrimidine], a significantly blocked SNP-induced p38 kinase and caveolin-1 activation in a dose-dependent manner. Therefore, to determine whether Src kinase activation is associated with dedifferentiation and/or COX-2 expression and PGE(2) production. As expected, PP2 potentiated SNP-stimulated dedifferentiation, but completely blocked both COX-2 expression and PGE2 production. And also, levels of p-Caveolin and COX-2 protein expression were increased in SNP-treated primary chondrocytes and osteoarthritic and rheumatoid arthritic cartilage, suggesting that p-Caveolin may play a role in the inflammatory responses of arthritic cartilage. CONCLUSION: Our previously studies indicated that NO caused dedifferentiation and COX-2 expression is regulated by p38 kinase through caveolin-1 (1). Therefore, our results collectively suggest that Src kinase regulates NO-induced dedifferentiation and COX-2 expression in chondrocytes via p38 kinase in association with caveolin-1.