• Food Science and Preservation
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• v.24 no.6
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• pp.842-848
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• 2017

Hylotelephium erythrostictum is commonly used as a medicinal herb. In this study, H. erythrostictum leaf (HEL), branch (HEB), root (HER), and above ground (HEAG) extracts were evaluated for their antioxidant properties. The antioxidant activities were assayed by three methods based on scavenging of DPPH, ABTS and superoxide anion radical. HEAG extract showed the highest DPPH, ABTS, superoxide anion radical scavenging activities. HEAG extract also exhibited the highest phenolic content (230 mg/g gallic acid equivalent). In our research for anti-inflammatory ingredients, the extract of HEAG inhibited the generation of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. To test the inhibitory effects of HEAG on pro-inflammatory cytokines, we conducted ELISA assay for the measuring the generation of tumor necrosis factor $(TNF)-{\alpha}$, IL (interleukin)-$1{\beta}$, and IL(interleukin)-6 in LPS-stimulated RAW264.7 macrophage cells. In these assays, HEAG ethanol extract showed a dose-dependent decrease in the production of $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6. Based on these results, extract of HEAG could be the efficient candidate for anti-inflammatory agents.

• Korean Journal of Plant Resources
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• v.25 no.5
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• pp.578-585
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• 2012

Cytokines released from innate immune cells play key roles in the regulation of the immune response. Red Ginseng (RG, steamed and dried root of Panax ginseng C.A. Meyer) is known to show different pharmacological effects by changed composition of saponins compared with Panax Ginseng. In this study, we examined the immunomodulatory effects of RG on the regulation of cytokine release in mice. RG was injected i.p at doses of 0.5, 5 and 50 mg/kg for 6 weeks. We assessed that the weight index of immune organs such as thymus, and spleen, and the mitogen blastogenesis of splenocytes. We also determined the levels of circulating cytokines in serum from RG-treated mice using ELISA assay. The weight index of thymus and spleen, and proliferation of mitogen response of splenocytes have increased in the RG-injected groups. In addition, the levels of IFN-${\gamma}$, TNF-${\alpha}$, IL-6, IL-12 and IL-2 concentrations have significantly increased in the serum of RG-treated mice, but that of IL-10 has not. These results suggest that RG has immune stimulating effects and could be useful as a immunoregulator of circulating cytokine release in vivo.

• Molecules and Cells
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• v.40 no.3
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• pp.222-229
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• 2017

Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

• Journal of Korean Medicine Rehabilitation
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• v.18 no.2
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• pp.17-31
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• 2008

Objectives : This study was carried out to investigate the effects of Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) on the monosodium iodoacetate-induced osteoarthritis in rats. Methods : Osteoarthritis was induced by injection of monosodium iodoacetate intraarticularly in both knee joints. Arthritic rats were divided into control and treated group. Control group were taken distilled water for 20days. Treated group were taken extracts of Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) by orally for the same duration. Normal group were injected normal saline and taken distilled water. Body weights were measured at 0, 5th, 10th, 15th, 20th day after injection. At the end of the experiment, gross and histopathological examination on the articular cartilages of the knee joints were performed. Proteoglycan contents of articular cartilages were analyzed by safranine O staining method. The contents of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 in synovial fluids were analyzed by ELISA method. Results : 1. Body weights of the treated group were significantly increased compared with control at 20days after injection. 2. Grossly, the severity of osteoarthritis in the treated group were alleviated compared with control. 3. Histopathologically, degenerative and necrotic lesion of articular cartilages in the treated group were alleviated compared with those of the control and histopathological scores of treated group were significantly decreased compared with control. 4. PG contents in articular cartilages of the treated group were significantly increased compared with control. 5. $TNF-{\alpha}$ contents in synovial fluids of the treated group were significantly decreased compared with control. Conclusions : According to above results, Samgi-eum($S{\bar{a}}nq{\grave{i}}-y{\check{i}}n$) has anti-arthritic effects on the monosodium iodoacetate-induced osteoarthritis in rats. And it is related with reduced secretion of $TNF-{\alpha}$ from osteoarthritic chondrocytes and synovial membranes.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.394-400
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• 2014

Avascular necrosis of the femoral head (ANFH) is commonly observed in patients treated with excessive glucocorticoid (GC). Single administration of lipopolysaccharide (LPS) has shown to induce immune stimulatory factors. However, the effect of repeated administration of LPS on GC-induced ANFH has not been studied. Thus, the purpose of this study was (i) to examine the cytokine profile induced by repeated LPS administrations and (ii) to test the effect of repeated LPS treatments on GC-induced ANFH. A mouse necrosis model of ANFH was designed by chronic GC administration with co-treatment of LPS. Mice body weights in the LPS/prednisolone (PDN) co-treated group were lower than that of the untreated control group, but spleen weights were greater than the control group. The levels of IL-6, $TNF{\alpha}$, and IL-33 in the liver and spleen of the LPS/PDN group were lower than the untreated control group, whereas $TNF{\alpha}$ level in the femoral head of the LPS/PDN group increased. Collectively, the effect of repeated LPS on the pathogenesis of GC-induced ANFH was associated with the $TNF{\alpha}$ level in the femoral head, but the pathogenesis did not correspond to cytokine levels in immune tissues.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.26 no.2
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• pp.45-57
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• 2013

Objectives : Daucus carota sativa has been frequently used as food supplements in many of the Asian countries, and a nutritional medical drug in traditional medicine. This research investigated the effects of Daucus carota sativa extract (DCE) on acute phases of inflammation in Raw 264.7 cells treated with lipopolysaccharide (LPS) in terms of the inhibition of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines production. Methods : NO, $PGE_2$, tumor necrosis factor (TNF)-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6 contents were assayed by ELISA, and expressions of inflammation-related proteins such as inducible NO synthase (iNOS) were determined by immunoblot analyses. Results : DCE treatment attenuated the LPS ability to increase the productions of NO and $PGE_2$ as well as the protein level of iNOS in a concentration-dependent manner. Consistently, treatment of the cells with DCE suppressed the production of TNF-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6. DCE also caused decreases of inhibitor of ${\kappa}B{\alpha}$ phosphorylation induced by LPS in the cells, which means DCE inhibition of NF-${\kappa}B$ activity. Furthermore, DCE blocked LPS-induced phosphorylation of p38 and SAPK/JNK. Conclusion : This study showing here may be of help to understand the action mechanism of DCE, and provide the information for the medical use of Daucus carota sativa for the inflammatory disease.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.77-81
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• 2007

Diagnostic performance of polymerase chain reaction (PCR) for detecting Dirofilaria immitis in dogs was evaluated when no gold standard test was employed. An enzyme-linked immunosorbent assay test kit (SnapTM, IDEXX, USA) with unknown parameters was also employed. The sensitivity and specificity of the PCR from two-population model were estimated by using both maximum likelihood using expectation-maximization (EM) algorithm and Bayesian method, assuming conditional independence between the two tests. A total of 266 samples, 133 samples in each trial, were randomly retrieved from the heartworm database records during the year 2002-2004 in a university animal hospital. These data originated from the test results of military dogs which were brought for routine medical check-up or testing for heartworm infection. When combined 2 trials, sensitivity and specificity of the PCR was 96.4-96.7% and 97.6-98.8% in EM and 94.4-94.8% and 97.1-98% in Bayesian. There were no statistical differences between estimates. This finding indicates that the PCR assay could be useful screening tool for detecting heartworm antigen in dogs. This study was provided further evidences that Bayesian approach is an alternative approach to draw better inference about the performance of a new diagnostic test in case when either gold test is not available.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.89-97
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• 2020

This study was carried out to examine a novel inactivated Salmonella Typhimurium (S. Typhimurium) vaccine candidate for protection of mice against salmonellosis by immunization of BALB/c mice using various type adjuvant. The novel type-inactivated vaccine candidate was constructed by adding Chlorhexidine digluconate solution. BALB/c mice were divided into 6 groups of 15 mice apiece. The mice were intramuscularly (IM) primed at 6 weeks of age and were IM boosted 8 weeks of age. Groups A and B mice were injected with sterile phosphate-buffered saline as controls; group C mice were inoculated with 5×10⁸ cells/100 µL of formalin-inactivated S. Typhimurium cells and adjuvant ISA70; groups D~F mice were immunized with 5×10⁸ cells/100 µL of the inactivated vaccine candidate and adjuvant ISA70, adjuvant IMS1313 and adjuvant IMS1313 containing 30 ㎍/mL of GI24, respectively. All mice (except group A mice) were orally challenged with a virulent S. Typhimurium strain at 10 weeks of age. Mice from groups C-F had significantly increased IgG levels compared to control groups (A-B) mice. The levels of splenocyte IFN-γ and IL-4 in mice of all groups were measured by ELISA, resulting in increased immunity in group F mice compared to those of groups A-E mice. These data suggested that systemic and cell-mediated immune responses were highly induced by IM immunization with the vaccine candidate and adjuvant IMS1313 containing GI24. Furthermore, clinical signs such as death were observed in only 20% of group F mice after virulent Salmonella strain challenge, however, groups B and C (100%), and groups D and E (60%) mice died. This data suggested that mice immunized by intramuscular prime and booster with this vaccine candidate and adjuvant IMS1313 containing GI24 effectively protected mice from salmonellosis.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.3
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• pp.33-55
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• 2007

Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.3
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• pp.109-124
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• 2007

Objectives In the present study, the author intended to investigate whether Gamitonggyu-tang (GTT) significantly affects (since the subject is GTT, you need an 's') in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of an airway, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2 for 3 weeks. Effects of orally-administered GTT for 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using enzyme-linked immunosorbent assay (ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GTT to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed.Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase (LDH) release. Also, the effect of GTT on contractility of isolated tracheal smooth muscle was investigated. Results (1) GTT inhibited hypersecretion of in vivo mucin. However, it did not affect the increase the number of goblet cells (2) GTT significantly increased mucin release from cultured HTSE cells, without significant cytotoxicity (3) GTT chiefly affected the 'mucin' secretion and did not affect the secretion of the other releasable glycoproteins with less molecular weight than mucin (4) GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle.Conclusions This result suggests that GTT can increase mucin secretion during short-term treatment (in vitro) whereas it can inihibit hypersecretion of mucin during long-term treatment (in vivo). The author suggests that the effect GTT with their components should be further investigated and it is valuable to find from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway epithelial cells.