• Restorative Dentistry and Endodontics
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• v.27 no.1
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• pp.1-11
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• 2002

Immune responses associated with bacterial infection involve various inflammatory cells. Clinical symptoms and pathologic features are particularly influenced by the predominant cells Among inflammatory cells, T cells have the heterogenity. T cells may develop into the mature cells expressing the cell surface markers with different functions and T helper cells are categorized into Th1 and Th2 cells based on their different patterns of cytokine production. The objective of this study was to investigate the change of expression of surface markers on T cells and the Th1/Th2 immune response in pulpal inflammation associated with specific bacteria. We experimentally induced pulpal inflammation in rat incisors by drilling without coolant and innoculated with Streptococcus mutans (S.M. group), Porphyromonas endodontalis (P.E. group), or only sterile cotton (control group). After 1, 2, and 5 days, mandibular incisors were extracted and the pulp tissues were extirpated The expressions of IL-2 recepters (CD25) and ICAM-1 (CD54) on CD4+ and CD8+ cells in the pulps were determined using a flow cytometer, and the concentration of IL-2, IFN-$\gamma$ and IL-4 was measured by enzyme-linked immunosorbent assay. The results were as follows: 1 In the S.M. group, CD4+ cells were more increased at 2nd day than 1st day and in the P.E. group, CD8+ cells were more increased at 2nd day than 1st day. 2. The percentages of CD4+, CD4+25+ and CD4+54+ cells were decreased in the pulp tissues at 5th day after irritation in all groups. 3. The ratios of CD4+/CD8+, CD4+/CD4+25+ and CD4+/CD4+54+ in the pulps at 2nd day after irritation by P. endodontalis were significantly lower than the other groups. 4. The higher concentrations of IFN-$\gamma$ than IL-4 in the pulps at 2nd day after irritation by P. endodontalis showed that T helper 1 reaction were predominant in the early stage of the pulpal inflammation induced by P. endodontalis. 5. The higher concentrations of IL-4 than IFN-$\gamma$ in the pulps at 1st day and 5th day after irritation by S. mutans were measured but the differences were not significant.

• Journal of Food Hygiene and Safety
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• v.3 no.2
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• pp.65-73
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• 1988

T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

• Journal of Periodontal and Implant Science
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• v.45 no.3
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• pp.101-110
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• 2015

Purpose: Sclerostin, an inhibitor of Wnt/${\beta}$-catenin signaling, exerts negative effects on bone formation and contributes to periodontitis-induced alveolar bone loss. Recent studies have demonstrated that serum sclerostin levels are increased in diabetic patients and that sclerostin expression in alveolar bone is enhanced in a diabetic periodontitis model. However, the molecular mechanism of how sclerostin expression is enhanced in diabetic patients remains elusive. Therefore, in this study, the effect of hyperglycemia on the expression of sclerostin in osteoblast lineage cells was examined. Methods: C2C12 and MLO-Y4 cells were used in this study. In order to examine the effect of hyperglycemia, the glucose concentration in the culture medium was adjusted to a range of levels between 40 and 100 mM. Gene expression levels were examined by quantitative reverse transcription-polymerase chain reaction and Western blot assays. Top-Flash reporter was used to examine the transcriptional activity of the ${\beta}$-catenin/lymphoid enhanced factor/T-cell factor complex. Tumor necrosis factor-alpha ($TNF{\alpha}$) protein levels were examined with the enzyme-linked immunosorbent assay. The effect of reactive oxygen species on sclerostin expression was examined by treating cells with 1 mM $H_2O_2$ or 20 mM N-acetylcysteine. Results: The high glucose treatment increased the mRNA and protein levels of sclerostin. High glucose suppressed Wnt3a-induced Top-Flash reporter activity and the expression levels of osteoblast marker genes. High glucose increased reactive oxygen species production and $TNF{\alpha}$ expression levels. Treatment of cells with $H_2O_2$ also enhanced the expression levels of $TNF{\alpha}$ and sclerostin. In addition, N-acetylcysteine treatment or knockdown of $TNF{\alpha}$ attenuated high glucose-induced sclerostin expression. Conclusions: These results suggest that hyperglycemia increases sclerostin expression via the enhanced production of reactive oxygen species and $TNF{\alpha}$.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.2
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• pp.56-71
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• 2014

Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.91-97
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• 2000

Objective: The presence of the various cytokines in human peritoneal fluid has been evaluated incompletely. Changes in cytokine levels may be related to activation of peritoneal macrophage and T-lymphocyte, development of endometriosis, and infertility. This study assesses peritoneal fluid levels of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in infertile women with endometriosis and normal women without endometriosis. Design: Prospective and case-control study in university hospital. Materials and Methods: Cytokine levels in peritoneal fluid obtained during laparotomy or laparoscopy from 21 patients in infertile patients with endometriosis and 24 controls undergoing laparotomy or laparoscopy with no evidence of pelvic endometriosis were determined by enzyme-linked immunosorbent assay. Results: The mean levels of interleukin-6 in infertile patients with endometriosis and controls were $72.7{\pm}23.7$ pg/ml and $18.5{\pm}9.7$ pg/ml respectively (p=0.02). Similarly, the mean levels of interleukin-8 in infertile patients with endometriosis was significantly higher than that of controls ($445.0{\pm}89.6$, vs $45.1{\pm}48.4$, p=0.04). The mean concentration of interleukin-10 in infertile patients with endometriosis was significantly lower than that of controls ($1.09{\pm}0.04$ vs $2.19{\pm}0.03$, p=0.03). The level of tumor necrosis factor-${\alpha}$ was not significantly different between the two study groups. Conclusions: Increased IL-6 and IL-8 and decreased IL-10 levels in the peritoneal fluid may be related to pathogenesis in the endometriosis and infertility, suggesting that partially contribute to the disturbed immune regulation observed in infertili women with endometriosis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.2
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• pp.241-246
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• 2004

To develop novel anti-aging peptides from the germinated black rice, we treated with bromelain, papain and Pronase E. And we investigated the effects of the germinated black rice peptide (GBRP) as anti-aging cosmetic ingredients, and compared with the non-germinated black rice protein (NBRP). We investigated the effects on in vitro inhibition of matrix-metalloprotease (MMP), proliferation of human skin fibroblasts, stimulation of collagen synthesis and expression of UVA-induced MMPs in human skin fibroblasts, UVA induced MMP-1 expression and collagen contents in human skin fibroblasts were analyzed by enzyme-linked immunosorbent assay (ELISA). As a result, the molecular weight distributions of GBRP and NBRP were determined by gel permeation chromatography to be approximately 900 and 10,000 daltons. GBRP increased skin cell proliferation about 40% and reduced UVA-induced MMP-1 expression about 50%. Also the collagen protein level of cells, which were cultured with GBRP, was increased about 25%. These results suggest that the geminated plant seed peptides can be novel anti-aging ingredients for cosmetics.

• 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.

• Journal of Neurogastroenterology and Motility
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• v.26 no.1
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• pp.106-116
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• 2020

Background/Aims Emerging evidence shows that the mechanism of irritable bowel syndrome (IBS) is associated with neurotrophic factors and tight junction proteins (TJPs). It is known that there are sex differences in the pathophysiology of IBS. The aim of the present study is to determine expression levels of neurotrophic factors, TJPs, and cytokines according to IBS subtype and sex. Methods From 59 IBS (33 IBS-constipation, 21 IBS-diarrhea, and 5 IBS-mixed) and 36 control patients, colonic mucosa mRNA expression levels of transient receptor potential vanilloid-1 (TRPV1), nerve growth factor (NGF), glial cell-derived neurotrophic factor (GDNF), and various TJPs were assessed by real-time polymerase chain reaction. Western blot was performed to determine levels of zonular occludens-1 (ZO-1). Serum levels of cytokines were measured by enzyme-linked immunosorbent assay. Results TRPV1, GDNF, and NGF mRNA levels were significantly increased in those with IBS-constipation compared to those in controls (all P < 0.05). However, they showed no significant difference between those with IBS-diarrhea and controls. Expression level of TRPV1 correlated with that of GDNF (r = 0.741, P < 0.001) and NGF (r = 0.935, P < 0.001). ZO-1 RNA expression levels were lower (P = 0.021) in female IBS-diarrhea than those in controls, although they showed no significant differences between male IBS-diarrhea and controls. Serum IL-1β levels in female IBS were significantly higher than those of male IBS, especially in IBS-constipation (P < 0.001). Conclusion Our results suggest that neurotrophic factors and IL-1β are closely related to IBS-constipation and that decrease of ZO-1 is an important factor in female with IBS-diarrhea.

• Korean Journal of Acupuncture
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• v.39 no.2
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• pp.43-53
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• 2022

Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.