The mast cell is one of the major effector cells in inflammatory reactions and can be found in most tissues throughout the body. Activated mast cells can produce histamine, as well as a wide variety of other inflammatory mediators such as eicosanoids, proteoglycans, proteases, and several pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-αα, and interleukins (IL-6), IL-8, IL-4, IL-13. In the present study, we isolated two bacterial strains (J80 and G147) from fermented soybean and Jeotgal, and investigated the inhibitory effects of their extracts which were prepared by several pretreatment methods (sonication for 20 min, heating at 100∘C100∘C for 30 min, autoclaving at 121∘C121∘C for 15 min) on the mast cell-mediated inflammatory response. The pretreated bacterial extracts had no cytotoxicity against Human Mast Cell (HMC-1). Among various pretreatments, the extracts treated at 100∘C100∘C showed highest inhibition of histamine release (J80, 28.46%; G147, 41.14%). The J80 and G147 extracts treated at 100∘C100∘C resulted in the inhibition of IL-6 secretion by 38.46% and 56.45%, respectively. The J80 extract treated at 100∘C100∘C resulted in the inhibition of TNF-αα secretion by 66.67%, but G147 extract showed the highest inhibition effect by 41.1% when treated with sonication. These results suggest that bacterial extracts treated at 100∘C100∘C have a higher level of anti-inflammatory effects than other treatments such as sonication or autoclaving.
Kim, Young-Kyoon;Kim, Seung-Joon;Park, Yong-Keun;Kim, Seok-Chan;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak;Kim, Sang-Ho
Tuberculosis and Respiratory Diseases
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v.49
no.6
/
pp.691-702
/
2000
Background : Acute lung injury (ALI) is a commonly encountered respiratory disease and its prognosis is poor when the treatment is not provided promptly and properly. However no specific pharmacologic treatment is currently available for ALI, although recently several supportive drugs have been under scrutiny. We studied anti-inflammatory effects of pentoxifylline (PF), a methylated xanthine, and ONO-5046, a synthetic neutrophil elastase inhibitor on lipopolysaccharide (LPS)-induced ALI in vitro. Methods : To establish an in vitro model of LPS-induced ALI, primary rat alveolar macrophages and peripheral neutrophils in various ratios (1:0, 5:1, 1:1, 1:5, 0:1) were co-cultured with transformed rat alveolar epithelial cells (L2 cell line) or vascular endothelial cells (IP2-E4 cell line) under LPS stimulation. Each experiment was divided into five groups-control, LPS, LPS+PF, LPS+ONO, and LPS+PF+ONO. We compared LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils in various ratios, and the resultant cytotoxicity on L2 cells or IP2-E4 cells between groups. In addition we also compared the productions of tumor necrosis factor (TNF)-αα interleukin (IL)-1β1β, monocyte chemotactic protein(MCP)-1, IL-6, and IL-10 as well as mRNA expressions of TNF-αα inducible nitric oxide synthetase(iNOS), and MCP-1 from LPS-stimulated primary rat alveolar macrophages between groups. Results : (1) PF and ONO-5046 in each or both showed a trend to suppress LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils regardless of their ratio, except for the LPS+PF+ONO group with the 1:5 ratio, although statistical significance was limited to a few selected experimental conditions. (2) PF and ONO-5046 in each or both showed a trend to prevent IP2-E4 cells from LPS-induced cytotoxicity by primary rat alveolar macrophages and peripheral neutrophils regardless their ratio, although statistical significance was limited to a few selected experimental conditions. the effects of PF and/or ONO-5046 on LPS-induced L2 cell cytotoxicity varied according to experimental conditions. (3) PF showed a trend to inhibit LPS-induced productions of INF-αα MCP-1, and IL-10 from primary rat alveolar macrophages. ONO-5046 alone didnot affect the LPS-induced productions of proinflammatory cytokines from primary rat alveolar macrophages but the combination of PF and ONO-5046 showed a trend to suppress LPS-induced productions of INF-αα and IL-10 PF and ONO-5046 in each or both showed a trend to increase LPS-induced IL-ββ and IL-6 productions from primary rat alveolar macrophages. (4) PF and ONO-5046 in each or both showed a trend to attenuate LPS-induced mRNA expressions of TNF-αα and MCP-1 from primary rat alveolar macrophages but at the same time showed a trend increase iNOS mRNA expression. Conclusion : These results suggest that PF and ONO-5046 may play a role in attenuating inflammation in LPS-induced ALI and that further study is needed to use these drugs as a new supportive therapeutic strategy for ALI.
Kim, Min-Ji;Bae, Nan-Young;Choi, Hyeun-Deok;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Sung, Nak-Yun;Byun, Eui-Hong;Nam, Hee-Sup;Ahn, Dong-Hyun
Microbiology and Biotechnology Letters
/
v.45
no.2
/
pp.101-109
/
2017
This study investigated the effect of the dichloromethane fraction form Katsuwonus pelamis heart on anti-inflammatory responses in lipopolysaccharide-stimulated RAW 264.7 cells and mouse models. Ethanol extract was partitioned with dichloromethane, ethyl acetate, butanol, and water. Among the fractions, the dichloromethane fraction showed a significant decrease in nitric oxide (NO) and pro-inflammatory cytokines [interleukin (IL)-6, IL−1βIL−1β, and tumor necrosis factor−αfactor−α] production compared to ethanol extract. The dichloromethane fraction attenuated the expression of inducible nitric oxide synthase and nuclear factor−κBfactor−κB (NF−κBNF−κB) p65 proteins in a dose-dependent manner. In addition, the expression of phosphorylation of mitogen-activated protein kinases (MAPKs) was also inhibited by the dichloromethane fraction. Moreover, the administration of 10, 50, and 250 mg/kg body weight-dose dependently inhibited the formation of edema by croton-oil and the application of dichloromethane (2 mg/ear) significantly reduced epidermal and dermal thickness and the infiltrated mast cell numbers. Therefore, the dichloromethane fraction exhibited an anti-inflammation effect by inhibiting NF−κBNF−κB and MAPK signaling activation in macrophages.
Background : Accumulating evidence shows that interleukin(IL)-1 plays a critical role in inflammation and connective tissue destruction observed in both osteoarthritis and rheumatoid arthritis. IL-1 induces gene expression related to cytokines, chemokines and matrix metalloproteinases by activation of many different transcription factors. Materials and Methods : The chondrosarcoma cell line, SW1353, is known to be a valuable in vitro system for investigating catabolic gene regulation by IL-1β1β in chondrocytic cells. To explore and analyze the changes in gene expression by IL-1 responsible for arthritis, SW1353 was treated with IL-1 for 1, 6 and 24 h and then total RNAs were purified for each time. The changes in gene expression were analyzed with 17k human cDNA microarrays and validated by semi-quantitative RT-PCR. Results : Greater than a two-fold change was observed in 1,200 genes including metallothioneins, matrix metalloproteinases, extracellular matrix proteins, antioxidant proteins, cytoskeleton proteins, cell cycle regulatory proteins, proteins for cell growth and apoptosis, signaling proteins and transcription factors. These changes appeared to be correlate with the pathophysiological changes observed in early osteoarthritis. Conclusion : cDNA microarray analysis revealed a marked variability in gene expression, and provided insight into the overall molecular changes. The result of this study provide initial information for further studies to identify therapeutic targets in osteoarthritis pathogenesis.
Objectives: This study was designed to examine the effects of extracts of Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) on the lipid lowering, anti-oxidation and concentration of proinflammatory cytokines and was investigated on hyperlipidemic rats. Methods: Male rats weighing 182.39±4.71g182.39±4.71g were fed high fat diet for 8 weeks and 36 rats(above 400 g) were divided into 4 groups. Each of 9 rats was divided a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) extracts(100 mg/kg, 200mg/kg, 300 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflmmatory cytokines, anti-oxidative activity and TNF−αTNF−α, Apo-B, Apo-E and leptin gene expression. Results: 1. Concentration of plasma free fatty(FFA) showed no significant difference in all the treatment groups. Concentration of plasma triglyceride(TG) showed a significant decrement in the 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups than that of control group. 2. Concentration of plasma total cholesterol showed a significant decrement in the 200 and 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups than that of control group. Concentration of plasma low density lipoprotein(LDL)-cholesterol showed a Significant decrement in the 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups than that of control group. Concentration of plasma high density lipoprotein(HDL)-cholesterol showed a significant increment in the 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) group. 3. Concentration of liver total cholesterol showed a tendence to decrease in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups. Concentration of liver TG showed a significant decrement in all Ojeoksangamibang groups than that of control group. 4. Concentration of plasma and liver thiobarbituric acid reactive substance(TBARS) showed a tendence to decrease in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups. 5. The values of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and catalase(CAT) activity showed a significant increment in all Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups than that of control group. 6. The values of plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity showed no significant different in all treatment group. 7. Concentration of plasma interleukin(IL)−1betainterleukin(IL)−1beta showed no significant difference in all the treatment groups. Concentration of plasma IL-6 showed a significant decrement in the 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) group than that of control group. Concentration of plasma tumor necrosis factor−α(TNF−α)factor−α(TNF−α) a siginifant decrement in the 200 and 300 mg/kg in Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) group than that of control group. However the concentration of plasma IL-10 in the 300 mg/kg Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups showed a significant increment than that of control group. 9. In the analysis of reverse transcription-polymerase chain reaction(RT-PCR), gene expression of TNF−αTNF−α, Apo-B and Apo-E in the Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups showed a lower expression than that of control group. However the gene expression of leptin showed no difference in the treatment groups. 10. The ratio of TNF−αTNF−α, Apo-B, and Apo-E per β−actinβ−actin expression in the Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) groups showed a significant decrement than that of control group. However The ratio of leptin expression per β−actinβ−actin expression showed no significant difference among all the treatment groups. Conclusions: According to above results, in lowering lipid effect, anti-oxidation and control of pro-inflammatory cytokines production, Ojeoksangamibang(WˇujˉisˇanjiˉawˊeifˉangWˇuj¯isˇanji¯aw`eif¯ang) gives effect.
The purpose of this study was to investigate the immuno-stimulatory activity of the high-molecular-weight fraction (HMWF) of Cynanchum wilfordii (CW) extracts obtained by ultrafiltration in murine macrophage RAW 264.7 cells and to assess its immuno-stimulatory effect in mice. Ultrafiltration was performed with polyethersulfone membranes (30 kDa cutoff) in a cross-flow filtration system to obtain the HMWF of CW. The results showed that the HMWF increased the production of various cytokines such as tumor necrosis factor-αα, interleukin-6, and nitric oxide in dose-ependent manners. In addition, HMWF treatment increased the relative spleen weight as well as splenocyte proliferation induced by concanavalin A or bacterial lipopolysaccharide in mice. Natural killer (NK) cell activity in the HMWF-treated group was significantly increased compared to that in the control group. These results suggest that the HMWF of CW can support the immune system through secretion of macrophage cytokines, thereby enhancing NK cell activity and murine splenocyte proliferation.
Kim, Hyoung-Jin;Kwon, O Jun;Lee, Ah Reum;Roh, Seong-Soo;Seo, Young-Bae
Journal of Applied Biological Chemistry
/
v.59
no.3
/
pp.179-188
/
2016
This study is aimed to evaluate the protective effect of Gastrodiae rhizoma and steamed, dried & fermented Gastrodiae rhizoma on Lipopolysaccharide (LPS)-induced hepatic injury in the mice model. Sample was selected to GR0F0 (not processed gastrodia rhizome) and GR6F4 (fermented with Saccharomyces cerevisiae before steamed and dried 6 times) based on 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid, and High-performance liquid chromatography analysis. Mice were randomly divided into 4 groups - Normal group, vehicle group (LPS treated), GR0F0 group (fed GR0F0 before LPS treated) and GR6F4 group (fed GR6F4 before LPS treated) with 6 mice in each group. GR0F0 group and GR6F4 group were fed each extract 200 mg/kg/day during 8 days. LPS 20 mg/kg injected to the experimental groups as abdominal injection. We measured aspartate aminotransferase, alanine amino-transferase in serum. GR0F0 and GR6F4 showed a significant decrease compared to the vehicle group. As a result of measuring the ROS, GR6F4 group showed a significant reduction in both the serum and liver tissues compared to the vehicle group. GR0F0 group showed a significant reduction only in the liver tissues. Activator protein-1, cyclooxygenase-2, and Inducible nitric oxide synthase were significantly decreased GR0F0 group and GR6F4 group. But tumor necrosis factor alpha only showed a significant reduction in GR6F4 group. GR0F0 and GR6F4 groups against liver damage in mice with LPS. That showed significant effects on anti-oxidant and anti-inflammatory action. The effects of GR6F4 group showed superior results compared to GR0F0 group. Therefore, Steamed, dried & fermented Gastrodia rhizoma was might have a protective effect on liver injury.
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.3
/
pp.335-341
/
2013
Kyungokgos purchased in local markets in Korea vary in their combination and mixing ratios during processing. This study was investigated qualities of Kyungokgos manufactured traditionally to evaluating its qualities. The general components of Kyungokgos were moisture (18.62~49.78%), ash (0.198~1.211%), protein (0.89~3.58%), lipid (0.16~1.14%) and carbohydrates (47.95~77.08%). The color values of L, a, and b were 26.49~73.87, 16.51~38.64, and 45.41~88.94, respectively. The viscosity was classified into three non-Newtonian type groups: high, medium, and non-dilatant, according to the increase of loop execution times. Three extracts (KOG-1, -7, and -8, in a 30-fold dilution) showed no cytotoxicity toward RAW 264.7 cells, while the extracts of KOG-2, -4, and -5 showed a low cytotoxic effect. KOG-1 and -2 extracts with low cytotoxicity markedly inhibited the production of the inflammatory mediators-nitric oxide (NO) and tumor necrosis factor-alpha (TNF-αα) in LPS-stimulated RAW 264.7 cells. These results indicate that KOG-1 and -2 extracts have anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophages.
As a research of inflammation inhibitory activity using natural resource, the inflammation inhibitory activity by purified active compound from Rhododendron mucronulatum flower was experimented. Rhododendron mucronulatum flower components were purified and separated with Sephadex LH-20 and MCI gel CHP-20 column chromatography, Purified compound was confirmed as myricetin by 1H−NMR1H−NMR, 13C−NMR13C−NMR and Fast atom bombardment (FAB)-Mass spectrum to have inhibition activity on inflammatory factors secreted by Raw 264.7 cells in response to lipopolysaccharide stimulation. Myricetin inhibited nitric oxide (NO) expression in a concentration dependent manner, approximately 40% inhibition was observed at a concentration of 50μM50μM. The inhibition effect of myricetin on inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression was 20% and 80%, respectively, at a concentration of 25μM25μM. Myricetin also inhibited expression of the inflammatory cytokines, tumor necrosis factor (TNF)−α(TNF)−α, interleukin (IL)−1β(IL)−1β, IL-6 and prostaglandin E2(PGE2)E2(PGE2) in a concentration dependent manner; a concentration of 50μM50μM, 70%, 80%, 80% and 95% inhibition was observed, respectively. Therefore myricetin isolated from Rhododendron mucronulatum flowers is expected to have an anti-inflammatory effect in Raw 264.7 cell induced by lipopolysaccharides. The results can be expected myricetin from Rhododendron mucronulatum flower to use as functional resource for anti-inflammatory activity.
Purpose: The aim of this study is to report the efficacy of infliximab, a monoclonal antibody directed against tumor necrosis factor alpha which is used for both treatment of refractory pediatric Crohn disease (CD) and induction of remission. Methods: Among pediatric patients who were diagnosed with CD at Samsung Medical Center between March 2001 and August 2007, a total of 16 patients were given infliximab to treat conventional therapyresistant refractory CD and severe active CD for induction of remission. Patients needing maintenance therapy were treated with an infliximab infusion every 8 weeks, and fistulizing CD patients occasionally received the infusion upon the condition that a fistula developed. The efficacy of treatment was assessed by comparing the Pediatric Crohn Disease Activity Index (PCDAI), Hct, ESR, CRP, and serum albumin levels using paired t-test. Results: The male/female ratio was 13:3, and the median age was 13 years (range, 21 months~15 years). The patients included 7 cases of therapy-resistant refractory CD, 7 cases of severe active CD, and 2 cases of fistulizing CD. Mean PCDAI before infliximab therapy was 34.19±±14.96, and mean follow-up PCDAI within 2 to 4 weeks after the last infusion was significantly lower, at 6.88±±10.31 (p=0.000). Hematological markers such as ESR (p=0.000), serum albumin (p=0.016), and CRP (p=0.009) also improved significantly after infusion. Remission was achieved in 2 of 4 patients refractory to conventional therapy. Among 3 steroid-dependent patients, 2 were able to discontinue steroid therapy, and dose reduction was possible in 1 patient. Remission after top-down therapy without prior use of other immunomodulators was achieved in 6 weeks in all 7 of the patients who had severe CD. Nine of ten refractory fistulizing CD patients also showed improvement after infliximab therapy. Conclusion: Infliximab was effective in pediatric refractory CD for induction of remission and maintenance therapy, as well as in severe CD for top-down induction therapy. Furthermore, infliximab has contributed to steroid cessation and dose reduction. Long-term follow-up evaluation is needed to determine safety and efficacy of infliximab in the future.
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