• 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 Preventive Medicine and Public Health
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• v.40 no.4
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• pp.285-290
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• 2007

Objectives : Cases of human brucellosis in Korea have recently increased due to the increasing incidence of bovine brucellosis. The authors conducted this study to elucidate the status of brucellosis through seroepidemiologic study. Methods : We selected our study population from a high risk group. We conducted a questionnaire survey and obtained blood samples to determine the seroprevalence of brucellosis antibodies for 10 days in February, 2005. The titers of brucellosis were measured by the combination of standard tube agglutination test (STA) and enzyme-linked immunosorbent assay (ELISA) test. Results : Our study subjects comprised 1,075 cases: 971 livestock workers, 51 veterinarians, and 53 artificial inseminators. In the STA test, 27 cases (2.5%) had titers of greater than or equal to 1:20. Of 1,068 cases (7 cases were excluded due to previous brucellosis), 7 cases of brucellosis were diagnosed with titers of 1:160, giving a seroprevalence of brucellosis of 0.66%. The seroprevalence in the male group was 0.95%, and that of livestock workers, veterinarians, and artificial inseminators was 0.52%, 4.17%, and 0.00%, respectively. The Spearman's correlation coefficient between the positive rate of bovine brucellosis per capita and household and human brucellosis was 0.806 and 0.744, respectively. The concordance rate between the Korea National Institute of Health and the Gyeongsangbuk-do Institute of Health and Environment by the STA and ELISA tests was 94.7% and 100.0%, respectively. Conclusions : The study results indicated in higher seroprevalence rate among veterinarians than among livestock workers and artificial inseminators. Because veterinarians may be exposed to this high risk, effective working guidelines for veterinarians to guard against brucellosis must be developed. Moreover, more extensive epidemiologic research for laboratory workers and meat handlers is needed.

• Tuberculosis and Respiratory Diseases
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• v.75 no.5
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• pp.205-209
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• 2013

Background: We investigated whether prunetin significantly affects tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced MUC5AC mucin gene expression, production, inhibitory kappa B ($I{\kappa}B$) degradation and nuclear factor kappa B (NF-kB) p65 translocation in human airway epithelial cells. Methods: Confluent NCI-H292 cells were pretreated with prunetin for 30 minutes and then stimulated with TNF-${\alpha}$ for 24 hours or the indicated periods. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effect of prunetin on TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65 was investigated by western blot analysis. Results: We found that incubation of NCI-H292 cells with prunetin significantly inhibited mucin production and down-regulated the MUC5AC gene expression induced by TNF-${\alpha}$. Prunetin inhibited TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65. Conclusion: This result suggests that prunetin inhibits the NF-${\kappa}B$ signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production regulated by the NF-${\kappa}B$ signaling pathway.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1308-1316
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• 2008

Propionibacterium acnes is known to playa pivotal role in the pathogenesis of acne vulgaris. CBT-SL5 is one of the antimicrobial peptides from Enterococcus faecalis SL5, and it has shown antimicrobial activity against P. acnes. The aim of this study was to investigate the anti-inflammatory effect of CBT-SL5 on the inflammation induced by P. acnes in cultured human keratinocyes. Cultured human keratinocytes derived from neonatal foreskin were treated with heat-killed P. acnes to induce inflammation, and then various concentrations of CBT-SL5 were added to the P. acnes-treated keratinocytes. The mRNA expression and protein secretion of interleukin (IL)-8, an inflammation marker, was analyzed by real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. We also analyzed the nuclear factor-kappa B (NF-$\kappaB$) p65 translocation by performing immunofluorescent staining. P. acnes treatment up regulated the IL-8 mRNA expression in the keratinocytes, and this was brought about through both toll-like receptor (TLR)2 and TLR4. At the concentrations of 10, 50, and 100 ng/ml, CBT-SL5 significantly down regulated the P. acnes-induced IL-8 mRNA expression and protein production (p<0.05). At 6 hand 12 h of the treatment, CBT-SL5 significantly suppressed the P. acnes-induced IL-8 mRNA expression. Secretion of IL-8 protein was significantly reduced at 24 h. The functional inhibitory activity of CBT-SL5 was shown by CBT-SL5 suppressing the P. acnes-induced NF-$\kappaB$ translocation from the cytoplasm to the nucleus. These results demonstrated that CBT-SL5 suppressed the P. acnes-induced IL-8 expression in keratinocytes. Therefore, CBT-SL5 may be a novel anti-inflammatory treatment for acne.

• Journal of Veterinary Clinics
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• v.32 no.4
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• pp.289-294
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• 2015

Fucoidan which is sulfated polysaccharide extracted from brown seaweed has a wide variety of internal biological activities. The objectives of this study were to examine the effect of fucoidan on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to investigate whether this effect is involved in the expression of inducible nitric oxide synthase (iNOS) and the activation of activator portein-1 (AP-1). The levels of NO production and AP-1 activity in the culture supernatants from porcine PBMCs were measured by the enzyme-linked immunosorbent assay and the levels of iNOS and AP-1 mRNA were determined by real time polymerase chain reaction. Fucoidan in LPS-naïve PBMCs has no effects on the production of NO and activity of AP-1. Expressions of iNOS and AP-1 mRNA in LPS-naïve PBMCs were also not affected by treatment of fucoidan. However, NO production, AP-1 activity and expressions of iNOS and AP-1 mRNA were dramatically increased in PBMCs stimulated with LPS. Enhancing effects of NO production and AP-1 activity in PBMCs induced by LPS were reduced by addition of fucoidan. Fucoidan also inhibited an increase in expressions of iNOS and AP-1 mRNA in LPS-stimulated PBMCs. These results suggested that fucoidan exerts anti-inflammatory effect by down-regulating production of NO via suppressing expression of iNOS and activity of AP-1 in LPS-stimulated porcine PBMCs.

• 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.

• Molecules and Cells
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• v.40 no.2
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• pp.133-142
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• 2017

Previous studies have shown that bone marrow mesenchymal stromal cell (MSC) transplantation significantly improves the recovery of neurological function in a rat model of intracerebral hemorrhage. Potential repair mechanisms involve anti-inflammation, anti-apoptosis and angiogenesis. However, few studies have focused on the effects of MSCs on inducible nitric oxide synthase (iNOS) expression and subsequent peroxynitrite formation after hypertensive intracerebral hemorrhage (HICH). In this study, MSCs were transplanted intracerebrally into rats 6 hours after HICH. The modified neurological severity score and the modified limb placing test were used to measure behavioral outcomes. Blood-brain barrier disruption and neuronal loss were measured by zonula occludens-1 (ZO-1) and neuronal nucleus (NeuN) expression, respectively. Concomitant edema formation was evaluated by H&E staining and brain water content. The effect of MSCs treatment on neuroinflammation was analyzed by immunohistochemical analysis or polymerase chain reaction of CD68, Iba1, iNOS expression and subsequent peroxynitrite formation, and by an enzyme-linked immunosorbent assay of pro-inflammatory factors (IL-$1{\beta}$ and TNF-${\alpha}$). The MSCs-treated HICH group showed better performance on behavioral scores and lower brain water content compared to controls. Moreover, the MSC injection increased NeuN and ZO-1 expression measured by immunochemistry/immunofluorescence. Furthermore, MSCs reduced not only levels of CD68, Iba1 and pro-inflammatory factors, but it also inhibited iNOS expression and peroxynitrite formation in perihematomal regions. The results suggest that intracerebral administration of MSCs accelerates neurological function recovery in HICH rats. This may result from the ability of MSCs to suppress inflammation, at least in part, by inhibiting iNOS expression and subsequent peroxynitrite formation.

• Gut and Liver
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• v.12 no.6
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• pp.682-693
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• 2018

Background/Aims: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. Methods: The effects of $17{\beta}$-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. Results: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p<0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-${\kappa}B$, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. Conclusions: E2 acts through the estrogen receptor ${\beta}$ signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.

• The Journal of Korean Obstetrics and Gynecology
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• v.32 no.4
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• pp.1-13
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• 2019

Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.