• Bulletin of the Korean Chemical Society
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• v.33 no.9
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• pp.3048-3054
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• 2012

The use of aspirin is widely recommended for the prevention of heart attacks owing to its ability to inhibit platelet activation by irreversibly blocking cyclooxygenase 1. However, aspirin also affects the fibrinolytic and hemostatic pathways by mechanisms that are not well understood, causing severe hemorrhagic complications. Here, we investigated the ability of aspirin and aspirin metabolites to inhibit thrombin-activatable fibrinolysis inhibitor (TAFI), the major inhibitor of plasma fibrinolysis. TAFI is activated via proteolytic cleavage by the thrombin-thrombomodulin complex to TAFIa, a carboxypeptidase B-like enzyme. TAFIa modulates fibrinolysis by removing the C-terminal arginine and lysine residues from partially degraded fibrin, which in turn inhibits the binding of plasminogen to fibrin clots. Aspirin and its major metabolites, salicylic acid, gentisic acid, and salicyluric acid, inhibit TAFIa carboxypeptidase activity. Salicyluric acid effectively blocks activation of TAFI by thrombin-thrombomodulin; however, salicylates do not inhibit carboxypeptidase N or pancreatic carboxypeptidase B. Aspirin and other salicylates accelerated the dissolution of fibrin clots and reduced thrombus formation in an in vitro model of fibrinolysis. Inhibition of TAFI represents a novel hemostatic mechanism that contributes to aspirin's therapy-associated antithrombotic activity and hemorrhagic complications.

• Journal of Life Science
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• v.23 no.12
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• pp.1501-1508
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• 2013

This study investigated antioxidant activity and the lipid content of serum for the possible outcome of improving the activity of Ishige okamurae extracts in ovariectomized rats. The antioxidant effects of an Ishige okamurae water extract and an Ishige okamurae ethanol extract were measured by evaluating DPPH free radical scavenging activity and SOD-like activity. Fifty, seven-week old female Sprague Dawley rats were randomly assigned to five groups as follows: sham-operated rats (SHAM), ovariectomized rats (OVX-CON), ovariectomized rats that were treated with 17-beta-estradiol (200 ${\mu}g/kg/day$), and ovariectomized rats that were treated with Ishige okamurae extracts (50 mg/kg/day and 200 mg/kg/day, respectively). The diets were fed to the rats for seven weeks after ovariectomy. The antioxidant activities of the water and ethanol extracts of Ishige okamurae increased in a dose-dependent manner, and the ethanol extract was found to be higher than the water extract. Therefore, we examined the effect of an Ishige okamurae ethanol extract on total serum cholesterol, triglycerides, HDL-cholesterol, and LDL-cholesterol levels, and anti-platelet aggregation. The total-cholesterol and triglyceride content of the serum increased in the OVX-CON group compared to the SHAM group, but supplementation with the Ishige okamurae ethanol extract caused these factors to decrease. Notably, the serum LDL-cholesterol concentration in the supplemented 200 mg/kg/day Ishige okamurae ethanol extract group was significantly more reduced than it was in the OVX-CON group. In addition, the platelet aggregation ability was lower in the groups treated with Ishige okamurae than it was in the OVX-CON group. According to these results, the effects of Ishige okamurae extract on serum lipid content in ovariectomized rats were illuminated.

• Journal of Ginseng Research
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• v.35 no.1
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• pp.86-91
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• 2011

Ginsenoside (G)-F1 is an enzymatic metabolite generated from G-Rg1. Although this metabolite has been reported to suppress platelet aggregation and to reduce gap junction-mediated intercellular communication, the modulatory activity of G-F1 on the functional role of skin-derived cells has not yet been elucidated. In this study, we evaluated the regulatory role of G-F1 on the cellular responses of B16 melanoma cells. G-F1 strongly suppressed the proliferation of B16 cells up to 60% at 200 ${\mu}g/mL$, while only diminishing the viability of HEK293 cells up to 30%. Furthermore, G-F1 remarkably induced morphological change and clustering of B16 melanoma cells. The melanin production of B16 cells was also significantly blocked by G-F1 up to 70%. Interestingly, intracellular signaling events involved in cell proliferation, migration, and morphological change were up-regulated at 1 h incubation but down-regulated at 12 h. Therefore, our results suggest that G-F1 can be applied as a novel anti-skin cancer drug with anti-proliferative and anti-migration features.

• The Journal of Advanced Prosthodontics
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• v.3 no.3
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1661-1669
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• 2007

Gamma-aminobutyric acid (GABA) is a non-protein amino acid. It is well known for its role as an inhibitory neurotransmitter of developing and operating nervous systems in brains. In this study, a novel function of GABA in the healing process of cutaneous wounds was presented regarding anti-inflammation and fibroblast cell proliferation. The cell proliferation activity of GABA was verified through an MTT assay using murine fibroblast NIH3T3 cells. It was observed that GABA significantly inhibited the mRNA expression of iNOS, IL-$1{\beta}$, and TNF-${\alpha}$ in LPS-stimulated RAW 264.7 cells. To evaluate in vivo activity of GABA in wound healing, excisional open wounds were made on the dorsal sides of Sprague-Dawley rats under anesthesia, and the healing of the wounds was apparently assessed. The molecular aspects of the healing process were also investigated by hematoxylineosin staining of the healed skin, displaying the degrees of re-epithelialization and linear alignment of the granulation tissue, and immunostaining and RT-PCR analyses of fibroblast growth factor and platelet-derived growth factor, implying extracellular matrix synthesis and remodeling of the skin. The GABA treatment was effective to accelerate the healing process by suppressing inflammation and stimulating re-epithelialization, compared with the epidermal growth factor treatment. The healing effect of GABA was remarkable at the early stage of wound healing, which resulted in significant reduction of the whole healing period.

• Journal of Microbiology and Biotechnology
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• v.23 no.7
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• pp.1015-1022
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• 2013

We recently showed that polylysine, the polymer of lysines, retains anti-prion activity. Although the effectiveness of prion inhibition by polylysine was demonstrated with the regimen tolerated in mice, a determination of quantitative polylysine toxicity is necessary to precisely address the in vivo toxicity level of polylysine. In this communication, we report the results of body weight monitoring and hematologic tests performed in CD-1 mice that received two different tolerable dosages of polylysine for an either 5-day or 4-week period. We found that there was no significant alteration in overall serum chemistry, blood cell count, and body weight gain compared with controls. The only notable quantitative change with statistical significance was the decrease of platelet numbers in mice subchronically administered with polylysine. Our results suggest that polylysine is harmless in mice if administered for a short period, but administrations of polylysine in mice may require considerate attention for safety in future investigations as mice chronically receive tolerable doses of polylysine.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.3
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• pp.349-360
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• 2018

Autophagy has been studied as a therapeutic strategy for cardiovascular diseases. However, insufficient studies have been reported concerning the influence of vascular smooth muscle cells (VSMCs) through autophagy regulation. The aim of the present study was to determine the effects of VSMCs on the regulation of autophagy under in vitro conditions similar to vascular status of the equipped micro-tubule target agent-eluting stent and increased release of platelet-derived growth factor-BB (PDGF-BB). Cell viability and proliferation were measured using MTT and cell counting assays. Immunofluorescence using an $anti-{\alpha}-tubulin$ antibody was performed to determine microtubule dynamic formation. Cell apoptosis was measured by cleavage of caspase-3 using western blot analysis, and by nuclear fragmentation using a fluorescence assay. Autophagy activity was assessed by microtubule-associated protein light chain 3-II (LC-II) using western blot analysis. Levels of intracellular reactive oxygen species (ROS) were measured using $H_2DCFDA$. The proliferation and viability of VSMCs were inhibited by microtubule regulation. Additionally, microtubule-regulated and PDGF-BB-stimulated VSMCs increased the cleavage of caspase-3 more than only the microtubule-regulated condition, similar to that of LC3-II, implying autophagy. Inhibitory autophagy of microtubule-regulated and PDGF-BB-stimulated VSMCs resulted in low viability. However, enhancement of autophagy maintained survival through the reduction of ROS. These results suggest that the apoptosis of conditioned VSMCs is decreased by the blocking generation of ROS via the promotion of autophagy, and proliferation is also inhibited. Thus, promoting autophagy as a therapeutic target for vascular restenosis and atherosclerosis may be a good strategy.

• Journal of Life Science
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• v.20 no.1
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• pp.90-96
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• 2010

Menopause increases the onset of hypertension and heart disease. Whereas it increases the blood LDL-cholesterol, triglyceride and total-cholesterol levels, the HDL-cholesterol concentration is reduced. Accordingly, we examined the effect of internal organs of Todarodes pacificus (IOT) on improvement in blood flow ability and changes in serum lipid content by using ovariectomized rats. For this study, the following four groups of 9-week-old Sprague-Dawley strain female rats were evaluated over 6 weeks: normal rats (SHAM), ovariectomized rats (CON) and ovariectomized rats that were treated with IOT extracts. Ovary removal promoted platelet aggregability. However, IOT administration in the CON group after ovary excision resulted in a hinderance of coherence. The blood vessel passing time of ovariectomized rats was slower than the SHAM group. But the blood flow ability, which was slowed down for ovary removal, was improved by IOT administration. Serum triglyceride levels were significantly reduced by IOT administration. Moreover, blood HDL-cholesterol levels were reduced by ovary removal. However, IOT administration after ovary excision significantly increase HDL-cholesterol levels. The biological activity of IOT could be confirmed from these results. Moreover, IOT can be used in the development of functional foods which are meant to improve blood circulation and anti-platelet aggregation function. According to these results, we could know that IOT improved blood flow and anti-platelet aggregation. Therefore, it is expected that IOT can be used for the development of functional foods.

• Biomolecules & Therapeutics
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• v.19 no.2
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• pp.187-194
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• 2011

Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.

• Journal of Life Science
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• v.19 no.9
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• pp.1245-1250
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• 2009

Beimu (Fritillaria thunbergii Miquel), a bulbous plant of Liliaceae found in Korea, Japan and China, has been used as an antitussive and expectorant agent, and is also useful in alleviating stonsillitis and bronchiolitis. Most researches have been focused on micro-propagation and plant regeneration, component analysis, and dormancy relieving of beimu. Reports regarding the biological activity of beimu, such as anti-Helicobacter pyroli or platelet aggregation inhibition activity, are few and not widely available. In this study, methanol extract and its organic solvent fractions were prepared from Fritillaria thunbergii Miquel and their antimicrobial, antithrombin, and antioxidant activities were evaluated, respectively. The methanol extract contained lots of water-soluble materials (58.98%) and hexane-soluble oils (14.85%). The ethylacetate and butanol fraction at $500{\mu}g$/disc concentration showed strong antibacterial activity against tested bacteria, except Escherichia coli. Antifungal activity was not observed in methanol extract and its fractions. The hexane, ethylacetate and butanol fractions showed strong antithrombin activity at 4.8 mg/ml concentration. Especially, the ethylacetate fraction showed 95.4 sec of thrombin time at a concentration of 1.2 mg/ml, which is comparable to aspirin, a widely used antithrombosis agent. For antioxidation activity, the ethylacetate and butanol fraction showed good 1,1-diphenyl-2-picryl hydrazyl scavenging activity ($IC_{50}$ of $344{\sim}368{\mu}g$/ml). In superoxide dismutase-like activity and reducing power, the fractions showed $20{\sim}25%$ of vitamin C, and $51{\sim}54%$ of butyl hydroxytoluene, respectively.