• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.4
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• pp.345-354
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• 2000

TGF-${\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to wound healing. The objective of this study is to investigate production of TGF-${\beta}_1$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-${\beta}_1$ which may be responsible for wound healing. The fibroblasts were originated from facial dermis and hypertrophic scar in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$) respectively, cells($5{\times}10^3ml$) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells($2.5{\times}10^5ml$) were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and LPS($0.1{\mu}g$) and SEB($0.1{\mu}g$) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-${\beta}_1$ was assayed in duplicate. The results were as follows. 1. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-${\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. 2. In hypertrophic scar fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation did not occur at 1 day after incubation, compared with the control. In SEB and LPS exposure in combination, the production of TGF-${\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. However, the production of TGF-${\beta}_1$ did not occur in SEB and LPS exposure respectively. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-${\beta}_1$ very significantly and both fibroblasts have different phenotype each other in this regard. This data suggest that the significant production of TGF-${\beta}_1$ may develope abnormal wound healing associated with tissue fibroproliferative disorder, such as hypertrophic scar and keloid formation.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.2
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• pp.137-146
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• 1999

Interactions among dexamethasone, dehydroepiandrosterone (DHEA), lipopolysaccharide (LPS), and antimycin A on the glutamate uptake and the polyamine uptake were investigated in primary cultures of rat cerebral cortical astrocytes to examine the effects of dexamethasone and DHEA on the regulatory role of astrocytes in conditions of increased extracellular concentrations of glutamate or polyamines. 1. $[^3H]Glutamate$ uptake: LPS and antimycin A decreased $V_{max},$ but both drugs had little effect on $K_m.$ Dexamethasone also decreased basal $V_{max}$ without any significant effect on $K_m.$ And dexamethasone further decreased the antimycin A-induced decrease of $V_{max}.$ DHEA did not affect the kinetics of basal glutamate uptake and the change by LPS or antimycin A. 2. $[^{14}C]Putrescine$ uptake: LPS increased $V_{max},$ and antimycin A decreased $V_{max}.$ They showed little effect on $K_m.$ Dexamethasone decreased $V_{max}$ of basal uptake and further decreased the antimycin A-induced decrease of $V_{max},$ and also decreased $V_{max}$ to less than control in LPS-treated astrocytes. DHEA did not affect $K_m$ and the change of $V_{max}$ by LPS or antimycin A. 3. $[^{14}C]Spermine$ uptake: Antimycin A decreased $V_{max},$ and LPS might increase $V_{max}.\;K_m$ was little affected by the drugs. Dexamethasone decreased basal $V_{max}$ and might further decrease the antimycin A-induced decrease of $V_{max}.$ And dexamethasone also decreased $V_{max}$ to less than control in LPS-treated astrocytes. DHEA might increase basal $V_{max}$ and $V_{max}$ of LPS-treated astrocytes. 4. $V_{max}$ of glutamate uptake by astrocytes was increased by putrescine (1000 ${\mu}M$ & 2000 ${\mu}M$) and spermidine (200 ${\mu}M,$ 500 ${\mu}M$ & 2000 ${\mu}M$). Spermine, 200 ${\mu}M$ (and 100 ${\mu}M$), also increased $V_{max},$ but a higher dose of 2000 ${\mu}M$ decreased $V_{max}.\;K_m$ of glutamate uptake was not significantly changed by these polyamines, except that higher doses of spermine showed tendency to decrease $K_m$ of glutamate uptake. In astrocytes, dexamethasone inhibited the glutamate uptake and the polyamine uptake in normal or hypoxic conditions, and the polyamine uptake might be stimulated by LPS and DHEA. Polyamines could aid astrocytes to uptake glutamate.

• Fisheries and Aquatic Sciences
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• v.20 no.12
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• pp.32.1-32.7
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• 2017

Recent in vitro studies have demonstrated that extract of soft coral Dendronephthya gigantea (SCDE) had strong anti-inflammatory activities. However, the direct effects of SCDE on anti-inflammatory activities in vivo model remained to be determined. Therefore, the present study was designed to assess in vivo anti-inflammatory effect of SCDE using lipopolysaccharide (LPS)-stimulated zebrafish model. We also investigated whether SCDE has toxic effects in zebrafish model. The survival, heart beat rate, and developmental abnormalities were no significant change in the zebrafish embryos exposed to at a concentration below $100{\mu}g/ml$ of SCDE. However, lethal toxicity was caused after exposure to 200 and $400{\mu}g/ml$ of SCDE. Treating zebrafish model with LPS treatment significantly increased the reactive oxygen species (ROS) and nitric oxide (NO) generation. However, SCDE inhibited this LPS-stimulated ROS and NO generation in a dose-dependent manner. These results show that SCDE alleviated inflammation by inhibiting the ROS and NO generation induced by LPS treatment. In addition, SCDE has a protective effect against the cell damage induced by LPS exposure in zebrafish embryos. This outcome could explain the profound anti-inflammatory effect of SCDE both in vitro as well as in vivo, suggesting that the SCDE might be a strong anti-inflammatory agent.

• The Korea Journal of Herbology
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• v.21 no.4
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• pp.69-76
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• 2006

Objectives : The fresh young leaves and dried fruits of Zanthoxylum piperitum (Korean name: Chopi) are used as diuretics, stomachies, anthelmintic and for the treatments of disorders of the digestive organ in Asia. We investigated inhibitory effects of Zanthoxylum piperitum extract on lipopolysaccharide(LPS)-induced production of nitric oxide(NO) and pro-inflammatory cytokines including $TNF-{\alpha}$ and $IL-1{\beta}$ from RAW264.7 mouse macrophage cells. Methods : After methanol extract of Zanthoxylum Fructus (Zanthoxylum extract) was pretreated in RAW264.7 cells, the cells were stimulated with LPS. Cell toxicity of Zanthoxylum extract was assayed bv MTT assay. The production of NO from the cells was measured in culture medium by Griess reaction. The production of $TNF-{\alpha}$ and $IL-1 \;{\beta}$ from the cells was measured in culture medium by ELISA. Results : Zanthoxylum Fructus extract greatly inhibited the production of inflammatory mediators such as NO, $TNF-{\alpha}$ and $IL-1{\beta}$ from LPS-stimulated RAW264.7 cells. Conclusion : This result suggests that Zanthoxylum extract may have an anti-inflammatory effect through the inhibition of inflammatory mediators.

• Herbal Formula Science
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• v.16 no.2
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• pp.219-228
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• 2008

Objective : The present study was designed to investigate whether the water extract of the root of Scutellaria baicalensis could regulate lipopolysaccharide (LPS)-induced type 1 interferon. Methods : To evaluate of type 1 interferon inhibitory effect of the root of Scutellaria baicalensis, we examined type 1 interferon in LPS-stimulated RAW264.7 cells. Furthermore, Interleukin (IL)-10 and interferon regulatory factor (IRF) - 1, 7 expression level were examined to study the inhibition mechanisms. Results 1. Extract from the root of Scutellaria baicalensis didn't have any cytotoxity itelf. 2. Extract from the root of Scutellaria baicalensis inhibited interferon-a,b in dose dependant- and type 1 interferon production in time dependant manner. 3. Extract from the root of Scutellaria baicalensis reduced IL-10 and IRF-1, 7 production in LPS-stimulated RAW 264.7 cells. Conclusion : The extract from the root of Scutellaria baicalensis down-regulated LPS-induced type 1 interferon through suppression of IL-10 and IRF-1, 7 expression. This results suggested that the extract from the root of Scutellaria baicalensis may be a beneficial drug against inflammatory diseases.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.114-114
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• 2022

Excessive endogenous endotoxin, especially lipopolysaccharide (LPS) reflux from gastrointestinal (GI) tract to the liver tissue is one of the most serious reasons of severe and acute liver injury which is mainly mediated by Kupffer cell activations. However, there is no clear molecular clues to explain the exact pathophysiological mechanism and effective drugs available till nowadays. We aimed to comprehend the pathophysiological features of LPS-induced liver injury and evaluate the efficacies of potential therapeutic drug, Ga-mi-Yuk-Mi-Jihwang-Tang (GYM), which is composed of herbal plants. GYM remarkably caused to normalize hepatic inflammation and oxidations against LPS-induced liver injury by evidence of serum liver enzymes, histopathological analysis, both hepatic protein and gene expression levels of pro-inflammatory cytokines, nitric oxide levels, and hepatic tissue levels of reactive oxygen species (ROS) levels, malondialdehyde (MDA), and 4-hydroxyneoneal, respectively. To assess molecular events in the hepatic tissue, we further found hepatic Sirtuin6 (Sirt6) levels were considerably depleted by LPS injection with aberrant alterations of Nrf2/HO-1 signaling pathways, whereas administration with GYM notably exerted to normalize these abnormalities. Our results exhibited that GYM would be one of target drug to diminish hepatic inflammation as well as oxidative stress by regulation of hepatic Sirt6 levels.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.5
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• pp.397-402
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• 2014

Microglia are activated by inflammatory and pathophysiological stimuli in neurodegenerative diseases, and activated microglia induce neuronal damage by releasing cytotoxic factors like nitric oxide (NO). Activated microglia synthesize a significant amount of vitamin $D_3$ in the rat brain, and vitamin $D_3$ has an inhibitory effect on activated microglia. To investigate the possible role of vitamin $D_3$ as a negative regulator of activated microglia, we examined the effect of 25-hydroxyvitamin $D_3$ on NO production of lipopolysaccharide (LPS)-stimulated microglia. Treatment with LPS increased the production of NO in primary cultured and BV2 microglial cells. Treatment with 25-hydroxyvitamin $D_3$ inhibited the generation of NO in LPS-activated primary microglia and BV2 cells. In addition to NO production, expression of 1-${\alpha}$-hydroxylase and the vitamin D receptor (VDR) was also upregulated in LPS-stimulated primary and BV2 microglia. When BV2 cells were transfected with 1-${\alpha}$-hydroxylase siRNA or VDR siRNA, the inhibitory effect of 25-hydroxyvitamin $D_3$ on activated BV2 cells was suppressed. 25-Hydroxyvitamin $D_3$ also inhibited the increased phosphorylation of p38 seen in LPS-activated BV2 cells, and this inhibition was blocked by VDR siRNA. The present study shows that 25-hydroxyvitamin $D_3$ inhibits NO production in LPS-activated microglia through the mediation of LPS-induced 1-${\alpha}$-hydroxylase. This study also shows that the inhibitory effect of 25-hydroxyvitamin $D_3$ on NO production might be exerted by inhibiting LPS-induced phosphorylation of p38 through the mediation of VDR signaling. These results suggest that vitamin $D_3$ might have an important role in the negative regulation of microglial activation.

• Biomolecules & Therapeutics
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• v.21 no.1
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• pp.21-28
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• 2013

Microglia play a role in maintaining and resolving brain tissue homeostasis. In pathological conditions, microglia release pro-inflammatory cytokines and cytotoxic factors, which aggravate the progression of neurodegenerative diseases. Autophagy pathway might be involved in the production of pro-inflammatory cytokines and cytotoxic factors in microglia, though details of the mechanism remain largely unknown. In the present study, we examined the role of the autophagy pathway in activated BV2 microglia cells. In BV2 cells, rapamycin treatment activated the formation of anti-LC3-labeled autophagosomes, whereas the ATG5 depletion using siRNA-ATG5 prevented the formation of LC3-labeled autophagosomes, indicating that BV2 cells exhibit an active classical autophagy system. When treated with LPS, BV2 cells expressed an increase of anti-LC3-labeled dots. The levels of LC3-labeled dots were not suppressed, instead tended to be enhanced, by the inhibition of the autophagy pathway with siRNA-ATG5 or wortmannin, suggesting that LPS-induced LC3-labeled dots in nature were distinct from the typical autophagosomes. The levels of LPS-induced expression of iNOS and IL6 were suppressed by treatment with rapamycin, and conversely, their expressions were enhanced by siRNA-ATG5 treatment. Moreover, the activation of the autophagy pathway using rapamycin inhibited cell death of LPS-stimulated microglia. These results suggest that although microglia possess a typical autophagy pathway, the glial cells express a non-typical autophagy pathway in response to LPS, and the activation of the autophagy pathway suppresses the expression of iNOS and IL6, and the cell death of LPS-stimulated microglia.

• Journal of Acupuncture Research
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• v.29 no.3
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• pp.41-53
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• 2012

Objectives : This study was designed to evaluate the effects of Akebiae Lignum herbal acupuncture(AL-HA) at $KI_{10}$ in acute nephritis induced by lipopolysaccharide(LPS) in rat. Methods : Rats were divided into 5 groups and 4 groups were injected LPS to induce acute nephritis. Normal group was normal SD rat, LPS group was injected LPS, AL-HA group was treated with AL-HA at $KI_{10}$ three times for a week, needle prick(NP) group with 26 gauge needle and saline group with normal saline. To evaluate the effects of AL-HA at $KI_{10}$ on acute nephritis in rats, WBC, neutrophil in blood, BUN, TNF-${\alpha}$, CINC-1 in serum and urinary volume, total protein in urine, renal MPO were measured and renal tissue was analyzed. Results : AL-HA group significantly reduced WBC, neutrophil in blood, BUN in serum, total protein in urine and renal MPO. And AL-HA group reduced concentration of neutrophil on glomerulus than LPS group in histological analysis. Conclusions : AL-HA at $KI_{10}$ has a therapeutic effect on acute nephritis in LPS stimulated rat. Therefore, it is suggested that AL-HA at $KI_{10}$ may be an useful therapeutics for acute nephritis in clinical field after further researches.

• Biomolecules & Therapeutics
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• v.26 no.2
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• pp.157-166
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• 2018

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a common clinical syndrome of diffuse lung inflammation with high mortality rates and limited therapeutic methods. Diosmetin, an active component from Chinese herbs, has long been noticed because of its antioxidant and anti-inflammatory activities. The aim of this study was to evaluate the effects of diosmetin on LPS-induced ALI model and unveil the possible mechanisms. Our results revealed that pretreatment with diosmetin effectively alleviated lung histopathological changes, which were further evaluated by lung injury scores. Diosmetin also decreased lung wet/dry ratios, as well as total protein levels, inflammatory cell infiltration and proinflammatory cytokine (eg. $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6) overproduction in bronchoalveolar lavage fluid (BALF). Additionally, increased MPO, MDA and ROS levels induced by LPS were also markly suppressed by diosmetin. Furthermore, diosmetin significantly increased the expression of Nrf2 along with its target gene HO-1 and blocked the activation of NLRP3 inflammasome in the lung tissues, which might be central to the protective effects of diosmetin. Further supporting these results, in vitro experiments also showed that diosmetin activated Nrf2 and HO-1, as well as inhibited the NLRP3 inflammasome in both RAW264.7 and A549 cells. The present study highlights the protective effects of diosmetin on LPS-induced ALI via activation of Nrf2 and inhibition of NLRP3 inflammasome, bringing up the hope of its application as a therapeutic drug towards LPS-induced ALI.