• Journal of Life Science
• /
• v.16 no.5
• /
• pp.840-849
• /
• 2006

Severe brain injuries induced by toxin pose one of the most important problems on our health care because of their high morbidity and mortality, are implicated to leucocyte infiltration more premature or immature brain than mature brain. Chemokines are the induction meditators for infiltration of inflammatory cells to the inflammation sites. In order to study the mechanism of leucocyte infiltration, the expression of several chemokines, MCP-1, $MIP-1{\alpha}$ and MIP-2 was studied in lipopolysaccharide(LPS)-stimulated neonatal and adult brain. One week old Sprague-Dawley rats or adult male rats weighing 300-350 g were used for the experiment. After anesthetization, $1\;{\mu}l$ LPS (0.5 mg/ml) subsequently was injected in the right caudate nucleus of the brain with stereotaxic frame. Animals were sacrificed at 6 hours, 24 hours, and 72 hours after injection. The present study was carried out using RT-PCR for the mRNA and immunohistochemistry for the expression of the proteins. In the neonatal rat brain, prominent interstitial edema with significant accumulation of leukocytes was detected at 24 and 72 hours after LPS injection. A semiquantitative analysis of RT-PCR revealed that the MCP-1, $MIP-1{\alpha}$, and MIP-2 mRNA expression peaked at 24 hours in neonatal and adult rat brain. Neonatal rats showed about 2.6, 1.4, and 1.2 times more expression of the MCP-1, $MIP-1{\alpha}$, and MIP-2 than that of the adult rats in the brain tissue. Immunohistochemical analysis also showed that MCP-1 immunoreactivity was paralleled with the RT-PCR results. MCP-1 protein was significantly detected at 24 and 72 hours in the brain parenchyma. $MIP-1{\alpha}$protein was highly expressed at 24 hours. The results of leukocyte infiltration in H&E stain was parallelled with that of the immunohistochemistry. Chemokine proteins were markedly detected at 24 hours after injection of LPS and neutrophil influx into intraparenchymal was prominent at 24 hours. These results suggest that the leukocyte infiltration in the intracranial infection may be controlled by mechanisms influenced by chemokine producing cells in the central nervous system such as microglia, astrocyte and endothelial cell.

• Journal of Life Science
• /
• v.31 no.12
• /
• pp.1110-1119
• /
• 2021

The purpose of this study was to investigate whether the inflammatory response in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages could be promoted by particulate matter 2.5 (PM_2.5) stimulation. To this end, the levels of inflammatory parameters, reactive oxygen species (ROS) and inflammation-regulating genes were investigated in RAW 264.7 cells treated with PM_2.5 in the presence or absence of LPS. Our results showed that the production levels of pro-inflammatory mediators (nitric oxide and prostaglandin E₂) and cytokines (interleukin-6 and -1β) were significantly increased by PM_2.5 stimulation in LPS-treated RAW 264.7 cells, which was correlated with increased expression genes involved in their production. In addition, when LPS-treated RAW 264.7 cells were exposed to PM_2.5, nuclear factor-kappaB (NF-κB) expression was further increased in the nucleus, and the expression of inhibitor of NF-κB as well as NF-κB in the cytoplasm was decreased. These results suggest that the co-treatment of PM_2.5 and LPS further increases the activation of the NF-κB signaling pathway compared to each treatment alone, thereby contributing to the promotion of transcriptional activity of inflammatory genes. Furthermore, although the generation of ROS was greatly increased by PM_2.5 in LPS-treated RAW 264.7 cells, the NF-κB inhibitor did not reduce the generation of ROS. In addition, when the generation of ROS was artificially suppressed, the production of inflammatory mediators and the activation of NF-κB were both abolished. Therefore, our results suggest that the increase in the NF-κB-mediated inflammatory response induced by PM_2.5 in LPS-treated RAW 264.7 macrophages was a ROS generation-dependent phenomenon.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.5
• /
• pp.843-848
• /
• 2011

In Korea, China, and Japan, Paeonia lactiflora Pall (PL) has been used in the treatment of rheumatoid arthritis, hepatitis, and fever for more than 1200 years. It has been reported that PL has protective effects against $H_2O_2$-induced oxidative stress and LPS-induced liver inflammation. However cellular and molecular mechanism of PL protection against oxidative stress has not fully been elucidated. Here, we describe that the water-soluble extract of PL decreased $H_2O_2$-induced hepatotoxicity. This hepatoprotective effect of PL is reason to decrease the level of intracellular reactive oxygen species (ROS) and increase expression of heme oxygenase 1 (HO-1) and heat shock protein 72 (HSP72) which proteins are involved in protecting the cells from stress like as oxidative stress. We also elucidated that hepatoprotective effect of PL was abolished by knock down of HO-1 and HSP72 by siRNA. These results suggest that the increasing of HO-1 and HSP72 protein by PL treatment might be participated in hepatoprotective effect against oxidative stress such as $H_2O_2$.

• Biomolecules & Therapeutics
• /
• v.25 no.5
• /
• pp.535-544
• /
• 2017

Carnosol is a phenolic antioxidant present in rosemary (Rosmarinus officinalis). It is known for anti-inflammatory effects, analgesic activity and anti-cancer effects. However, no study has been dedicated yet to its effect on atopic dermatitis (AD). Here, we show that carnosol effectively inhibited LPS-induced nitric oxide (NO) generation and expression of inflammatory marker proteins (iNOS and COX-2) in RAW 264.7 cells. In addition, carnosol effectively inhibits the phosphorylation of STAT3 and DNA binding activity in RAW 264.7 cells. Pull down assay and docking model analysis showed that carnosol directly binds to the DNA binding domain (DBD) of STAT3. We next examined the anti-atopic activity of carnosol ($0.05{\mu}g/cm^2$) using 5% Phthalic anhydride (PA)-induced AD model in HR1 mice. Carnosol treatment significantly reduced 5% PA-induced AD like skin inflammation in skin tissues compared with control mice. Moreover, carnosol treatment inhibits the expression of iNOS and COX-2 in skin tissue. In addition, the levels of $TNF-{\alpha}$, $IL-1{\beta}$, and Immunoglobulin-E in blood serum was significantly decreased in carnosol treated mice compared with those of 5% PA treated group. Furthermore, the activation of STAT3 in skin tissue was decreased in carnosol treated mice compared with control mice. In conclusion, these findings suggest that carnosol exhibited a potential anti-AD activity by inhibiting pro-inflammatory mediators through suppression of STAT3 activation via direct binding to DBD of STAT3.

• Biomolecules & Therapeutics
• /
• v.26 no.5
• /
• pp.446-457
• /
• 2018

Adiponectin, a hormone predominantly originated from adipose tissue, has exhibited potent anti-inflammatory properties. Accumulating evidence suggests that autophagy induction plays a crucial role in anti-inflammatory responses by adiponectin. However, underlying molecular mechanisms are still largely unknown. Association of Bcl-2 with Beclin-1, an autophagy activating protein, prevents autophagy induction. We have previously shown that adiponectin-induced autophagy activation is mediated through inhibition of interaction between Bcl-2 and Beclin-1. In the present study, we examined the molecular mechanisms by which adiponectin modulates association of Bcl-2 and Beclin-1 in macrophages. Herein, we demonstrated that globular adiponectin (gAcrp) induced increase in the expression of AUF1 and ZFP36L1, which act as mRNA destabilizing proteins, both in RAW 264.7 macrophages and primary peritoneal macrophages. In addition, gene silencing of AUF1 and ZFP36L1 caused restoration of decrease in Bcl-2 expression and Bcl-2 mRNA half-life by gAcrp, indicating crucial roles of AUF1 and ZFP36L1 induction in Bcl-2 mRNA destabilization by gAcrp. Moreover, knock-down of AUF1 and ZFP36L1 enhanced interaction of Bcl-2 with Beclin-1, and subsequently prevented gAcrp-induced autophagy activation, suggesting that AUF1 and ZFP36L1 induction mediates gAcrp-induced autophagy activation via Bcl-2 mRNA destabilization. Furthermore, suppressive effects of gAcrp on LPS-stimulated inflammatory mediators expression were prevented by gene silencing of AUF1 and ZFP36L1 in macrophages. Taken together, these results suggest that AUF1 and ZFP36L1 induction critically contributes to autophagy induction by gAcrp and are promising targets for anti-inflammatory responses by gAcrp.

• Microbiology and Biotechnology Letters
• /
• v.43 no.2
• /
• pp.112-119
• /
• 2015

The anti-inflammatory effect of Sargassum coreanum ethanolic extract (SCEE) was investigated using lipopolysaccharide (LPS)-induced inflammatory responses in this study. It was shown that there was no cytotoxicity in the viability of macrophages treated with SCEE when compared to the control. The production of NO was considerably suppressed by SCEE, approximately up to 50% at 100 μg/ml. This significantly decreased levels of IL-6, TNF-α, and IL-1β. In addition, the expression of iNOS, COX-2, NF-κB was suppressed by SCEE treatment. In in vivo testing, the croton oil-induced mouse ear edema was attenuated by SCEE and there were no mortalities in mice administered with 5000 mg/kg body weight of SCEE over a 2 week observation period. From these results, SCEE inhibits the release of LPS-induced pro-inflammatory cytokines and mediators, suggesting that SCEE could be a potential agent for anti-inflammatory therapies.

• Journal of Life Science
• /
• v.24 no.4
• /
• pp.370-376
• /
• 2014

Although the grasshopper Oxya chinensis sinuosa has long been used as food in Korea, there is little data on its functional effects. In this study, we investigated the anti-inflammatory effect of O. c. sinuosa ethanol extract (OCE) in RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for induction of inflammation. First, we determined that there is no cytotoxicity at $2,000{\mu}g/ml$ or less of OCE in RAW 264.7 cells. To evaluate the anti-inflammatory effects of OCE, we investigated expression levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, and pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether OCE could inhibit translocation of NF-${\kappa}B$ p65 into the nucleus in LPS induced RAW 264.7 cells. As a result, we found that the mRNA and protein levels of TNF-${\alpha}$ and IL-6 decreased in LPS-induced RAW 264.7 cells after treatment with OCE in a dose-dependent manner. In addition, we confirmed a $2,000{\mu}g/ml$ concentration of OCE inhibited translocation of NF-${\kappa}B$ p65 by immunnostaining and Western blot analysis, and a decrease in the protein expression levels of iNOS and COX-2. Accordingly, we suppose that OCE has an anti-inflammatory effect through down-regulation of TNF-${\alpha}$, IL-6, iNOS, and COX-2 related to ${\kappa}B$ p65 inflammatory signaling pathways.

• Journal of Life Science
• /
• v.26 no.11
• /
• pp.1282-1288
• /
• 2016

Artemisia, a plant widely used as traditional herbal medicine in many countries, has drawn attention of the researchers. And its extracts or compounds are known to have an efficacy of antioxidant, anti-diabete, anti-cancer, anti-inflammation and neuroprotection. Sumaeyaksuk is a variant of the Artemisia argyi and major constituents are eupatilin and jaceosidin. This study was performed to investigate the effects of the sumaeyaksuk aqueous extract on inflammatory response induced by lipopolysaccharide (LPS) in human hepatoma HepG2 cells. To examine the potential hepatoprotective properties of sumaeyaksuk extract, cell viability, as well as nitric oxide (NO), reactive oxygen species (ROS), macrophage colony-stimulating factor (M-CSF), interleukin-8 (IL-8) levels, alanine transaminase (ALT), and aspartate transaminase (AST) activities, were measured. Cytotoxic activity of extracts on HepG2 cells was measured by MTT assay. Sumaeyaksuk extract did not induce cytotoxicity at concentrations of $0{\sim}400{\mu}g/mL$. NO and ROS levels significantly decreased with increasing concentration of the extract. The secretion levels of M-CSF and IL-8 were suppressed by sumaeyaksuk extract in a dose-dependent manner. Moreover, ALT (75.4%) and AST (61.6%) levels significantly decreased in sumaeyaksuk extract-treated cells at $400{\mu}g/mL$. These results suggested that the sumaeyaksuk extract attenuates the LPS-induced hepatotoxicity resulting from regulation of inflammatory factors and could potentially be used as a hepatitis therapeutic agent.

• Journal of Life Science
• /
• v.26 no.1
• /
• pp.117-122
• /
• 2016

In the current study, the ethanol extracts and their subsequent organic solvent fractions from lees of sweet potato soju were prepared and the prepared samples were designated as from KSD-E8-1 to KSD-E8-5. Their effects on cell viability and nitric oxide (NO) production in mouse macrophage RAW 264.7 cells were investigated. The results showed that the ethyl acetate fraction (KSD-E8-3) of lees extracts from sweet potato soju significantly decreased nitric oxide (NO) production in LPS-activated RAW 264.7 cells, whereas they did not affect cell viabilities. The fraction KSD-E8-3 reduced the expression of pro-inflammatory genes such as COX-2, iNOS and TNF-alpha and also decreased protein expression of iNOS in a dose dependent manner, which were detected with RT-PCR and Western blot analysis, respectively. In addition, we detected the expression of mitogen-activated protein kinases (MAPKs) such as p38, JNK, and ERK1/2 and their phosphorylated forms. The results indicated that the treatment of the fraction KSD-E8-3 did inhibit phosphorylation of p38, JNK, and ERK1/2 MAPKs, indicating that the fraction KSD-E8-3 regulates LPS-induced inflammatory response via suppressing MAPK signaling pathway. Overall, these results may contribute to understand the molecular mechanism of anti-inflammatory effects by the ethyl acetate fraction of lees extracts from sweet potato soju.

• Journal of Korean Medicine Rehabilitation
• /
• v.31 no.1
• /
• pp.33-46
• /
• 2021

Objectives The aim of this study is to evaluate anti-inflammatory and anti-arthritic effects of Sogyunghwalhyel-tang-gamibang (SGHHTGB) in cell and animal models and also to suggest one of putative mechanisms underlying its anti-arthritic effects. Methods Enzyme-linked immunosorbent assay was applied to measure the concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in culture medium and blood serum and nitric oxide (NO) was assayed by Griess reagent. The expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. Results In a cell model using RAW264.7 macrophages stimulated with the endotoxin lipopolysaccharide (LPS), the drug, at its non-cytotoxic concentrations, inhibited the production of the pro-inflammatory cytokine TNF-α, IL-1β and IL-6. In addition, it suppressed the expression of the inflammatory enzyme iNOS and COX-2, and reduced the synthesis of the enzyme product NO (as stable nitrite) and PGE2 in activated macrophages. Meanwhile, in an animal model using rheumatic arthritis (RA) mice induced with injection of type II collagen antibody (CAb) and LPS, the drug improved clinical symptom of arthritis and reduced paw thickness and inflammatory cell infiltration. In blood of RA mice, the drug reduced serum levels of TNF-α, IL-1β, IL-6, nitrite, and PGE2, all inflammatory mediators produced by activated macrophages. Conclusions SGHHTGB may ameliorate CAb and LPS-induced RA in mice, presumably by inactivating macrophages that are capable of initiating joint inflammation by producing pro-inflammatory cytokines and expressing inflammatory enzymes.