• Tuberculosis and Respiratory Diseases
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• v.77 no.2
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• pp.73-80
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• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

• Tuberculosis and Respiratory Diseases
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• v.77 no.2
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• pp.65-72
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• 2014

Background: It is valuable to find the potential activity of regulating the excessive mucin secretion by the compounds derived from various medicinal plants. We investigated whether aqueous extract of the root bark of Morus alba L. (AMA), kuwanon E, kuwanon G, mulberrofuran G, and morusin significantly affect the secretion and production of airway mucin using in vivo and in vitro experimental models. Methods: Effect of AMA was examined on hypersecretion of airway mucin in sulfur dioxide-induced acute bronchitis in rats. Confluent NCI-H292 cells were pretreated with ethanolic extract, kuwanon E, kuwanon G, mulberrofuran G, or morusin for 30 minutes and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 hours. The MUC5AC mucin secretion and production were measured by enzyme-linked immunosorbent assay. Results: AMA stimulated the secretion of airway mucin in sulfur dioxide-induced bronchitis rat model; aqueous extract, ethanolic extract, kuwanon E, kuwanon G, mulberrofuran G and morusin inhibited the production of MUC5AC mucin induced by PMA from NCI-H292 cells, respectively. Conclusion: These results suggest that extract of the root bark and the natural products derived from Morus alba L. can regulate the secretion and production of airway mucin and, at least in part, explains the folk use of extract of Morus alba L. as mucoregulators in diverse inflammatory pulmonary diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1427-1433
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• 2006

The antioxidant activity, nitrite scavenging effect, and angiotensine I-converting enzyme inhibiting activity of solvent extracts of soybean isoflavone were investigated. Also anticarcinogenic activities of them were examined by MTT assay using human cancer cell line. Isoflavone aglycones showed relatively high antioxidant activity in order of glycitein, genistein, and daidzein. Methanol extract of soybean actively scavenged free radicals and was shown excellent nitrite scavenging effect. Glycitein and methanol extract of soybean inhibited the growth of human cancer cell such as stomach carcinoma (SNU-1) and colon carcinoma (SNU-C4) effectively. Genistein, daidzein and methanol extracts of soybean inhibited the growth of cancer cell such as stomach carcinoma (SNU-1), but had weak activities to colon carcinoma (SNU-C4). To applicate the soybean isoflavone as an enhancer for food quality and processibility, the stabilities on heat and pH of isolated isoflavone, isoflavone in soybean flour and isoflavone concentrate of soybean were investigated. Stability of isoflavone concentrate of soybean was decreased in pH extreme (below 3, above 8) during sterilization, but isoflavone in soybean flour showed higher stability in all pH range. All kinds of isoflavone tested in this study were very stable during the heat treatment.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2993-2998
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• 2009

An enzyme-linked immunosorbent assay (ELISA)-on-a-chip (EOC) biosensor combined with cell concentration technology based on immuno-magnetic separation (IMS) was investigated for use as a potential tool for early screening of Listeria monocytogenes (L. monocytogenes) in food products. The target analyte is a well-known pathogenic foodborne microorganism and outbreaks of the food poisoning typically occur due to contamination of normal food products. Thus, the aim of this study was to develop a rapid and reliable sensor that could be utilized on a daily basis to test food products for the presence of this pathogenic microorganism. The sensor was optimized to provide a high detection capability (e.g., 5.9 ${\times}\;10^3$ cells/mL) and, to eventually minimize cultivation time. The cell density was condensed using IMS prior to analysis. Since the concentration rate of IMS was greater than 100-fold, this combination resulted in a detection limit of 54 cells/mL. The EOC-IMS coupled analytical system was then applied to a real sample test of fish intestines. The system was able to detect L. monocytogenes at a concentration of 2.4 CFU/g after pre-enrichment for 6 h from the onset of cell cultivation. This may allow us to monitor the target analyte at a concentration less than 1 CFU/g within a 9 h-cultivation provided a doubling time of 40 min is typically maintained. Based on this estimation, the EOC-IMS system can screen and detect the presence of this microorganism in food products almost within working hours.

• 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.

• Food Science and Preservation
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• v.14 no.4
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• pp.425-430
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• 2007

To investigate the potential therapeutic value of a plant extract against amyloid ${\beta}-peptide-induced$ cell damage, we first screened extracts of 250 herbs, and finally selected a water extract of Spinacia oleracea for further study. This extractshowed the potential to inhibit the reactions of oxidants. We measured the angiotensin-converting-enzyme (ACE) inhibitory activity of the extract, and assessed the ability of the extract to protect neuronal cells from chemical-induced cell death. SH-SY5Y neuroblastoma cells were used in this assay. The extract exerted protective effects on $H_2O_2-induced$ cell death, when $H_2O_2$ was used at 100 M, 200 M, and 500 M (protection of 87%, 73%, and 58%, respectively). When 50 M of amyloid ${\beta}-peptide$ was added to the test cells, however, the extract had no protective effect on cell death. The extract inhibited ACE activity in a dose-dependent manner, and exhibited potent protection against the deleterious effects of $H_2O_2$. In sum, these results suggest that a water extract of Spinacia oleracea has the potential to afford protection against chemical-induced neuronal cell death, and the extract may be useful in the treatment of neurodegenerative diseases. The precise molecular mechanism of neuroprotection is under investigation.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.581-590
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• 1998

We developed a novel water-soluble camptothecin analobue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respecitively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to imduce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/$ED-{58}$) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in <5% tumor bearing mouse. this schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.39
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• pp.8.1-8.8
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• 2017

Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.1
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• pp.57-77
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• 2011

Objectives : This study was to investigate the suppression effects of Sopunghwalhyeol-tang(Shufenghuoxie-tang) on the monosodium iodoacetate-induced osteoarthritis in rats. Methods : Arthritis was induced by injection of monosodium iodoacetate(0.5 mg) into the both knee joints of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken distilled water for 20 days. Treated group was taken extracts of Sopunghwalhyeol-tang(Shufenghuoxie-tang) by oraly for same duration. Normal group(n=8) was injected with normal saline and was taken distilled water for 20 days. Macroscopic examination and histopathological study on articular cartilage of knee joint were operated at 20 days after injection. Proteoglycan(PG) content of articular cartilages of knee joint was represented by safranine O staining, was measured at 20 days after injection. Tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$, in synovial fluid were measured with enzyme-linked immuno sorbent assay(ELISA) kit at 20 days after injection. Immunohistochemical staining of cyclo-oxygenase-2(COX-2), inducible nitric oxide synthase(iNOS) in knee joints were observed at 20 days after injection. Results : 1. Lymphocytes in peripheral blood the treated group was significantly decreased compared with the control group. 2. PG content in articular cartilage of the treated group was significantly increased compared with the control group. 3. Histopathologically, osteoarthritic scores of the treated group was significantly decreased compared with the control group. 4. $TNF-{\alpha}$ content in synovial fluid of the treated group was significantly decreased compared with the control group. 5. COX-2 revelation index in chondrocytes and synovial membrane of the treated group was significantly decreased compared with the control group. 6. Matrix metalloproteinase-3(MMP-3) revelation index in chondrocytes and synovial membrane of the treated group was significantly decreased compared with the control group. Conclusions : On the basis of these results, we concluded that Sopunghwalhyeol-tang(Shufenghuoxie-tang) has inhibiting effects on the $TNF-{\alpha}$, COX-2 and MMP-3 secretion of chondrocytes and synovial membrane in Monosodium Iodoacetate-induced osteoarthritis model of rats.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.54-60
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• 2020

Objective: Oxidative stress plays a key role in the pathogenesis of male infertility. But, the adverse effects of oxidative biomarkers on sperm quality remain unclear. This study aimed to investigate the levels of nitric oxide (NO), 8-hydroxydesoxyguanosine (8-OHdG), and total antioxidant capacity (TAC) oxidative biomarkers in seminal plasma and their relationship with sperm parameters. Methods: A total of 77 volunteers participated in the study, including fertile (n = 40) and infertile men (n = 37). NO, 8-OHdG, and TAC levels were measured using the ferric reducing ability of plasma, Griess reagent method and an enzyme-linked immunosorbent assay kit, respectively. Results: The mean values of sperm parameters in the infertile group were significantly lower than those in the fertile group (p< 0.001). The mean 8-OHdG in the seminal plasma of infertile men was significantly higher (p= 0.013) than those of controls, while the mean TAC was significantly lower (p= 0.046). There was no significant difference in NO level between the two groups. The elevated seminal 8-OHdG levels were negatively correlated with semen volume, total sperm counts and morphology (p< 0.001, p= 0.001 and p= 0.052, respectively). NO levels were negatively correlated with semen volume, total sperm counts and morphology (p= 0.014, p= 0.020 and p= 0.060, respectively). Positive correlations between TAC and both sperm count and morphology (p= 0.043 and p= 0.025, respectively) were also found. Conclusion: These results suggested that increased levels of NO and 8-OHdG in seminal plasma could have a negative effect on sperm function by inducing damage to the sperm DNA hence their fertility potentials. Therefore, these biomarkers can be useful in the diagnosis and treatment of male infertility.