• Korean Journal of Psychosomatic Medicine
• /
• v.8 no.1
• /
• pp.46-57
• /
• 2000

Objectives : The purpose of the present study was to examine the effect of subacute treatment with the selective serotonin reuptake inhibitors(paroxetine and sertraline) on immobility in the forced swim test(FST) and on FST-induced changes in immune parameters of the mice. Methods : Authors applied a modified method of FST by Porsolt et al. Over 5 BALB/c mice were used for each group of experiments. To explore the changes in immune parameters by FST, authors investigated the production of anti-rat RBC antibody, concanavalin A(ConA)- or lipopolysaccharide(LPS)-stimulated splenocytes proliferation assay and cytokine gene expression. Results : Both paroxetine and sertraline decreased the duration of immobility in a dose-related manner. FST-performed mice showed a significant decrease in mitogenic responses of splenocytes and a slight increasing tendency in anti-rat RBC antibody response. All these responses were attenuated significantly by paroxetine and attenuated nearly nominal significance level by sertraline. The cytokine profiles of ConA-stimulated splenocytes from FST-performed mice showed stronger expression of IL-4 and weaker expression of IL-2 than control mice, and no changes in the expressions of IFN-$\gamma$ and lymphotoxin. IL-6 and IL-10 were not expressed in both group of mice. The pretreatment of paroxetine and sertraline attenuated the altered cytokine expressions in FST-performed mice to some extent. Some alterations of the expressions of IL-6 and IL-10 were observed in the mice which the selective serotonin reuptake inhibitors had been pretreated. Conclusion : The subacute treatment of paroxetine and sertraline attenuated the FST-induced behavioral and immune changes, and these serotonin reuptake inhibitors may exert some modulating effects on the immune system by the induction of cytokine gene expression, especially IL-6 and IL-10.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.9
• /
• pp.1333-1339
• /
• 2011

After mulberry (Morus alba) leaves were fermented with Hericium erinaceum mycelium by solid-state culture to enhance physiological activity, fermented mulberry leaves (MA-HE) was extracted by hot-water (MA-HEHW) and ethanol (MA-HE-E). MA-HE-HW showed enhanced mitogenic and intestinal immune system modulating activities (1.41 and 1.52 fold of saline control, respectively) compared to hot-water extracts of non-fermented mulberry leaves (MA-HW) and H. erinaceum mycelium (HE-HW) at $100\;{\mu}g$/mL. Meanwhile, when we tested the inhibitory effects of extracts on nitric oxide (NO), tumor necrosis factor (TNF)-${\alpha}$, and interleukin (IL)-$1{\beta}$ and IL-6 production, MA-HE-E significantly inhibited these pro-inflammatory mediators in LPS-stimulated RAW 264.7 cells (45.1, 41.3, 70.2, and 55.7% inhibition of LPS control at $1,000\;{\mu}g$/mL). In addition, MA-HE-HW and MA-HE-E did not show any cytotoxicity on RAW 264.7 cells at $1,000\;{\mu}g$/mL whereas HE-E and MA-E indicated cytotoxicity (80.1 and 30.7% cell viability of saline control). These results suggest that mulberry leaves fermented with H. erinaceum by solid-state culture might have enhanced immunomodulatory and anti-inflammatory effects compared to non-fermented mulberry leaves, resulting in ingredients biotransformed for fermentation with H. erinaceum mycelium.

• Journal of Periodontal and Implant Science
• /
• v.26 no.2
• /
• pp.469-490
• /
• 1996

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.11
• /
• pp.1535-1542
• /
• 2009

To find the new use of Korean ginseng and mushroom, crude polysaccharides were prepared from submerged cultures of Hericium erinaceum in the medium supplemented with Korean ginseng extracts. When we fractionated crude polysaccharides (HE-GE-CP-1, 3, and 5) from hot-water extracts of submerged cultures of H. erinaceum with ginseng extracts (1%, 3%, and 5% addition of total medium), the yields of HE-GE-CP-1, 3, and 5 were identified at 5.7, 5.1, and 4.8%, respectively. Among crude polysaccharide fractions, HE-GE-CP-5 was significantly higher (1.89-fold of the saline control) than those of HE-GE-CP-1 (1.64-fold) or HE-GE-CP-3 (1.76-fold) on mitogenic activity of splenocytes. HE-GE-CP-5 also had the more potent bone marrow cell proliferation (1.83-fold) rather than HE-CP or HE-GE-CP-1 or HE-GE-CP-3 (1.59- or 1.44- or 1.69-fold, respectively), and anti-metastatic activity as anti-cancer effect showed the highest prophylactic value (72.4% inhibition of tumor control) in 5% supplementation of ginseng extract. However, the lysosomal phosphatase of macrophage was significantly stimulated after HE-GE-CP-3 treatment (2.03-fold). In addition, the immunostimulating and anti-metastatic crude polysaccharide, HE-GE-CP-5, contained mainly neutral sugars (63.2%) with considerable amounts of uronic acid (19.3%) and a small amount of proteins (8.8%). HE-GE-CP-5 can stimulate immune system to inhibit tumor metastasis, and its anti-tumor metastasis may be associated with macrophages, splenocytes and Peyer's patch cells activation.

• Tuberculosis and Respiratory Diseases
• /
• v.46 no.3
• /
• pp.338-349
• /
• 1999

Polymorphonuclear leukocytes(PMN) are the predominant inflammatory cells recruited in acute lung injury such as adult respiratory distress syndrome, pneumonia and also chronic lung disease such as idiopathic pulmonary fibrosis and pulmonary emphysema. Interleukin-8(IL-8) is an 8,000 D protein produced by many cells and has potent neutrophil chemoattractant and activating properties. The GRO, also called melanoma growth-stimulatory activity(MGSA), referring to a peptide of 73 amino acids, was reported to be mitogenic for cultured human melanoma cells. Mature GRO/MGSA has marked sequence similarity to IL-8. In view of the structural similarities to IL-8, it was of particular interest to test GRO for neutrophil activating and chemotactic properties. We found a significant release of IL-8 and GRO/MGSA from the cultured human umbilical vein endothelial cell(HUVEC) which was stimulated either with TNF$\alpha$ or IL-1$\beta$ and also found the expression of IL-8 and GRO/MGSA mRNA. Neutrophil chemotactic activity was enhanced in accordance with the increased IL-8 and GRO/MGSA. Our study also suggest that the IL-8 is more important in the increased neutrophil chemotactic activity than GRO/MGSA when endothelial cell is stimulated with TNF$\alpha$ or IL-1$\beta$ in vitro.

• Tuberculosis and Respiratory Diseases
• /
• v.58 no.4
• /
• pp.359-366
• /
• 2005

Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.

• Korean Journal of Food Science and Technology
• /
• v.42 no.2
• /
• pp.223-232
• /
• 2010

Crude polysaccharide (WG-PL-CP) was fractionated from fermented ginseng with Phellinus linteus in solid culture to enhance the immunostimulation of ginseng. WG-PL-CP produce three active polysaccharide-rich fractions (WG-PL-CP-II, III, and IV) on DEAE-Sepharose CL-6B ($Cl^-$ form). WG-PL-CP-III displayed higher mitogenic activity (1.98-fold of the saline control at $100\;{\mu}g/mL$) than did WG-CP-III or PL-CP-III (1.60- or 1.65-fold, respectively), and potent intestinal immune system modulating activity through Peyer's patch was obtained by WG-PL-CP-IV only (1.56-fold). Meanwhile, WG-PL-CP-II and III significantly enhanced macrophage stimulating activity (2.01- and 1.94-fold) compared to WG-CP-II and III (1.73- and 1.66-fold) or PL-CP-II and III (1.79- and 1.72-fold). In addition, WG-PL-CP-III and IV mainly contained neutral sugar (73.5 and 67.3%) and uronic acid (23.2 and 24.6%). Component sugar analysis also showed that WG-PL-CP-III consisted mainly of uronic acid as well as the neutral sugars Glc, Ara, Gal, Rha and Xyl (molar ratio of 0.81:1.00:0.49:0.42:0.28:0.20), whereas WG-PL-CP-IV was mainly comprised of uronic acid, Ara, Rha, Gal, Xyl and Glc (1.00:0.75:0.69:0.63:0.42:0.34). Therefore, it is assumed that these active polysaccharides play an important role in enhancing the immunostimulation of fermented ginseng with P. linteus in solid culture.

• Tuberculosis and Respiratory Diseases
• /
• v.49 no.3
• /
• pp.310-322
• /
• 2000

Background : Phospholipase C(PLC) plays an important role in cellular signal transduction and is thought to be critical in cellular growth, differentiation and transformation of certain malignancies. Two second messengers produced from the enzymatic action of PLC are diacylglycerol (DAG) and inositol 1, 4, 5-trisphosphate (IP3). These two second messengers are important in down stream signal activation of protein kinase C and intracellular calcium elevation. In addition, functional domains of the PLC isozymes, such as Src homology 2 (SH2) domain, Src homology 3 (SH3) domain, and pleckstrin homology (PH) domain play crucial roles in protein translocation, lipid membrane modificailon and intracellular memrane trafficking which occur during various mitogenic processes. We have previously reported the presence of PLC-${\gamma}1$, ${\gamma}2$, ${\beta}1$, ${\beta}3$, and ${\delta}1$ isozymes in normal human lung tissue and tyrosine-kinase-independent activation of phospholipase C-${\gamma}$ isozymes by tau protein and AHNAK. We had also found that the expression of AHNAK protein was markedly increased in various mstologic types of lung can∞r tissues as compared to the normallungs. However, the report concerning expression of various PLC isozymes in lung canærs and other lung diseases is lacking. Therefore, in this study we examined the expression of PLC isozymes in the paired surgical specimens taken from lung cancer patients. Methods : Surgically resected lung cancer tissue samples taken from thirty seven patients and their paired normal control lungs from the same patients, The expression of various PLC isozymes were studied. Western blot analysis of the tissue extracts for the PLC isozymes and immunohistochemistry was performed on typical samples for localization of the isozyme. Results : In 16 of 18 squamous cell carcinomas, the expression of PLC-${\gamma}1$ was increased. PLC-${\gamma}1$ was also found to be increased in all of 15 adenocarcinoma patients. In most of the non-small cell lung cancer tissues we had examined, expression of PLC-${\delta}1$ was decreased. However, the expression of PLC-${\delta}1$ was markedly increased in 3 adenocarcinomas and 3 squamous carcinomas. Although the numbers were small, in all 4 cases of small cell lung cancer tissues, the expression of PLC-${\delta}1$ was nearly absent. Conclusion : We found increased expression of PLC-${\gamma}1$ isozyme in lung cancer tissues. Results of this study, taken together with our earlier findings of AHNAK protein-a putative PLD-${\gamma}$, activator-over-expression, and the changes observed in PLC-${\delta}1$ in primary human lung cancers may provide a possible insight into the derranged calcium-inositol signaling pathways leading to the lung malignancies.