Objectives : Trimellitic anhydride (TMA), a sensitizer that induces occupational asthma and atopic dermatitis, is widely used industrially to make epoxy and alkyd resins, plasticizers, high temperature polymer, and surfactants. The aim of this study was to investigative the effects of aqueous extracts of Lonicera japonica Flower(LJFAE) on TMA-induced contact hypersensitivity (CHS) in Balb/c mice. Methods : The dried flowers of L. japonica were extracted with distilled water at $100^{\circ}C$ for 7 h. The extract was freeze-dried following filteration through 0.45 ${\mu}m$ filter. Mice were orally administrated with or without LJFE of a different doses(25-100 mg/kg) for 28 days. In the challenge period, mice were externally applied at difference doses of LJFAE one time per day 30 min before TMA treatment. We examined the effects of LJFAE on the the serum levels of IgE and prostagladin E2 (PGE2), the Thl/Th2 cytokine production of spleen cells, ear swelling responses, and the leukocyte infiltration induced by TMA. Results : The orally and externally administration of LJFAE dose-dependently reduced the serum levels of IgE and PGE2 production as well as ear swelling responses and leukocyte infiltration in TMA-induced Balb/c mice. Furthermore, the levels of Thl (TNF-${\alpha}$, IFN-${\gammer}$, IL-2)/Th2 (IL-4, IL-5, IL-13) cytokine production from spleen cells stimulated with anti-CD3 and CD28 mAbs was markedly suppressed by the orally and externally treatment with LJFAE in a concentration dependent manner. Conclusions : These results suggest that LJFAE suppresses the inflammatory mediators and regulates the Thl/Th2 cytokines. Therefore, these properties may contribute to the strong anti-CHS response effect of LJFAE.
Objectives Eucommiae cortex is one of the frequently used herbs for musculoskeletal disorders, with well-known anti-inflammatory effect. Meanwhile, powdered form of herbal drugs is more advantageous than decoction form, in that storing and taking is more convenient. Thus, the authors aimed to investigate whether the drying method affects the index compound level and anti-inflammatory effect of eucommiae cortex water extract. Methods Eucommiae cortex was extracted in distilled water at $100^{\circ}C$ for 3 hours, filtered, and then evaporated under vacuum. One half of the sample was freeze-dried at $-80^{\circ}C$. Another half of the sample was added with dextrin and then spray-dried at $180^{\circ}C$. To assess the possible change in index compound content, pinoresinol diglucoside was selected as the index marker. The content level of the index compound in various extract sample was quantified through high performance liquid chromatography. In addition, iNOS assay in LPS-induced RAW 264.7 cells was adopted to determine the anti-inflammatory effect of various extract samples. Furthermore, MTT assay was performed to confirm that the result of iNOS assay was not due to cytotoxicity. Results There was no significance difference in index compound content between extract samples obtained through two different drying methods. Anti-inflammatory activity of extract samples were similar at the matching concentration, regardless of the drying methods. Extract samples did not show any significant cytotoxicity. Conclusions Extract samples of eucommiae cortex were obtained through freeze-drying and spray-drying. Neither change in index compound content nor difference in anti-inflammatory activity was observed between drying methods.
The Langerhans cells are dendritic nonkeratinocytes found suprabasally in most stratified squamous epithelia, such as human epidermis and the epithelium of the oral mucosa including that of gingiva. After Paul Langerhans found it in the skin in 1968, there have been sturdies of it's function and distribution . Stingle et al. reported that the Langerhans cells seem able to present antigens and to stimulate T-lymphocytes. Shelley et al. discovered that they can take up contact allergens. Accordingly it has been suggested that Langerhans cells are important elements of p Peripheral cell mediated immune system. In this study, the gingival tissue of a adult periodontitis patient was taken and freeze dried. In one specimen, we used the CD1 monoclonal antbody to staining the Langerhans cell. The other specimen, we embedded in paraffin and staining it with S-100 monoclonal antibody. The purpose of this study was to use these specimens to find out the distribution, orientation, morphology of the Langerhans cell and to discover the increase or decrease of Langerhans cell in an increased inflammatory state. The results were obtained as follows : 1. Langerhans cells were distributed between the basal cell layer and spinous cell layer against the CD1 & S-100 monoclonal antibody. 2. Langerhans cessl were plentiful in the oral eptihelium, and there was very little in the sulcular epithelium. 3. There were no Langerhans cell in the junction epithelium and pocket lining epithelium. 4. The number of Langerhans cells that responsed to the CD1 & S-100 monoclonal antibody had a statistically difference. 5. As the infiltration of the lymphocyte into the connective tissue were increased, the number of Langerhans cells in the epithelium were increased. 6. As the inflammation was increased, Langerhans cells in the spinous cell layer were more increased than those of the basal layer.
This study was designed to investigate the effects of BojungIkkiTang-Gamybang freeze dried powder (BITG) on proliferauion, protective of cell death induced by chemicals such as paraquat, hydrogen peroxide etc and anti-oxidative effects in C6 glioma cells. In our results, BITC accelerated proliferation rates of C6 cells in vitro. In addition, protective effects on cell death induced by paraquat and hydrogen peroxide. And, BITC did not have effects on SOD and total glutathione activities, but decresed malone dialdehyde activity. In conclusion, these results suggest the possibility of BojungIkkiTang-Gamybang to protect brain cell or neuronal cell from damage induced by oxidative stress. And also suggest that related mechanisms are involved in malone dialdehyde activity.
Hot water soluble extract was prepared from Hericium erinaceus and its neuritogenic activity on PC12h cells was analyzed, which is a clone originating from a rat pheochromocytomon. The moisture content of freeze dried hot water extract was 12.08%. The extract was mainly composed of carbohydrate (51.24%) followed by crude protein (24.04%), crude fat (0.26%), dietary fiber (5.09), and ash (12.18%). Fatty acids, glucan and inorganic constituents were found as minor components. The neuritogenic activity of hot water extract was evaluated under microscopic observation of neurite outgrowth in PC12h cells and by measuring the neurite length of induvidual cell. The extract exhibited strong effect of neurite outgrowth in a dose-dependent manner from 0.01 mg/mL to 1 mg/mL, in which longer neurite outgrowth was observed as the treatment dose increased.
Shibyukmiyouki-Eum was a drug that treated carbuncle and cellulitis. So, the purpose of this study was to investigate effect of Shibyukmiyouki-Eum on the anticancer and inhibitive effects of the secondary effects by anticarcinogen We used Shibyukmiyouki-Eum extract(SYE) with freeze-dried, 8wks-old male mice(balb/c, ICR) and cancer cell lines(L1210. S-180) for this Study, The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). and measurement of WBC, RBC, hemoglobin and platelet was tested by a automated hematology analyzer. The results of this study were obtained as follow; SYE was showed significantly cytotoxicity on the L1210 and S-180 cell lines. increased proliferation of thymocytes decreased by anticarcinogen. In combined effects of SYE and vincristine(0.005㎎/kg), SYE was significantly inhibited proliferation of L1210 cells and significantly increased proliferation of thymocytes. Also SYE was significantly increased count of WBC. platelet and increased count of RBC, hemoglobin. These results suggest that SYE has not only anticancar action but inhibitive effects of on the secondary effects by anticarcinogen.
Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.
To repair bony defects with tansplanted bone in the body, fresh autogenous bone is undoubtly, the most effective bone graft for clinical applications. But the demineralized bone has the matrix-induced bone formation which was suggested by Urist in 1965. Many authors assisted that demineralized bone powder induces phenotypic conversion of mesenchymal cells into osteoblasts, with high-density bone formation. The process of inducing differentiated cells becomes osteogenic properties. The purpose of this study was to evaluate the osteoinductive capacity of allogenic freeze-dried demineralized bone block (FDD, $7{\times}7mm$) and to compare FDD with the same sue of deep-frozen allogenic bone(DF), fresh autogenous bone (A) after implantation. The histological and ultrastructural features of tissue responses were examined after 1, 2, 4, 6, 8 weeks implantation of each experimental groups in the operative site of the New Zealand white rabbits. The results were as follows : 1. Inflammatory cell infiltration generally has appeared at 1 week, but reduced at 4 weeks in each group, but most severe in DF group. 2. Osteoblastic activity has increased for 4 weeks, but decreased at 6 weeks in each group and there was no significant difference among experimental groups. 3. New bone formation has begun at 1week, least activations in A groups, and showed the revesal line of bone formation among each group at 6 to 8 weeks. 4. Bone resorption has appeared at 1 week, but disappeared at 4 weeks in both A and DF groups, but more severe in DF than A groups. 5. In ultrastructural changs, the DF group have showed the most remarkable osteoclastic activities among experimental groups. 6. Osteoid or tangled collagen fibrils near the implanted sites were replaced by more mature, lamellated bony trabeculae during bone remodeling. There was little difference among each experimental groups. 7. During the convertion osteoblasts to osteocytes which embedded within the bone matrix, there was organ-less-poor cytoplasm, increased nuclear chromatin, abundant rough endothelial reticulum (RER) in each groups. From the above the findings, the DF group shored more bone resorption and foreign body reaction than FDD and A groups, and FDD group showed more new bone formation or osteoblastic activity than DF and A groups in early stage. There was no significant difference of cellular activities among the FDD DF, and A groups according to the time.
Sunjeonhwadok-Tang was a drug that treated carbuncle and cellulitis. So, the purpose of this study was to investigate effects of Sunjeonhwadok-Tang on the proliferation of cancer calls and immunocytes focusing around combined effects of anticarcinogen. We used Sunjeonhwadok-Tang extract(SHT) with freeze-dried, 8wks-old male balb/c mice and cancer cell lines(L1210, Sarcoma-180) for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results : 1. SHT was significantly showed cytotoxicity on the L1210 cell lines. 2. SHT was significantly increased proliferation of thymocytes and splenocytes in vitro. 3. In combined effects of SHT and vincristine(0.005 mg/kg), SHT was significantly inhibited proliferation of L1210 cell lines, but was not inhibited proliferation of Sarcoma-180 cell lines compared with positive control group. 4. In combined effects of SHT and vincristine(0.005 mg/kg), SHT was significantly increased proliferation of thymocytes and splenocytes compared with positive control group. 5. In combined effects of SHT and vincristine, SHT was significantly increased proliferation of thymocytes and splenocytes in normal mice. 6. In combined effects of SHT and vincristine, SHT was significantly inhibited proliferation of L1210 cells in L1210 cells transplanted mice 7. In combined effects of SHT and vincristine, SHT was significantly increased proliferation of L1210 cell in L1210 cells transplanted mice. The present author thought that SHT had action of anti-cancer and immuno-activity, and in combined effects of vincristine, SHT had recoverable effects on damage by anticarcinogen.
This study investigated the protective effect of the butanol (BuOH) fraction from fermented Laminaria japonica extract (BFLJ) on AAPH-induced oxidative stress in porcine kidney epithelial cells (LLC-PK1 cells). L. japonica was fermented by Aspergillus oryzae at $35{\pm}1^{\circ}C$ for 72 h. Freeze-dried fermented L. japonica was extracted with distilled water, and the extracted solution was mixed with ethanol and then centrifuged. The supernatant was subjected to sequential fractionation with various solvents. The BuOH fraction was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. The BuOH fraction of fermented L. japonica had a protective effect against the AAPH-induced LLC-PK1 cells damage and increased cell viability while reducing lipid peroxidation formation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. The inhibitory effect of BFLJ on lipid peroxidation formation had a higher value of $0.11{\pm}0.01nmol$ MDA at $100{\mu}g/mL$ concentration in comparison with intact BuOH fraction showing $0.22{\pm}0.08nmol$ MDA at the same concentration. Furthermore, BFLJ treatment increased glutathione concentration. GSH concentration in the cell treated with BFLJ of $100{\mu}g/mL$ was $1.80pmol/L{\times}10^5cells$. These results indicate that BFLJ protects the LLC-PK1 cells against AAPH-induced cell damage by inhibiting lipid peroxidation formation and increasing antioxidant enzyme activities and glutathione concentration.
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