• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.7
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• pp.936-942
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• 2012

In the present study, the effects of green tea catechin on microsomal phospholipase $A_2$ ($PLA_2$) activity and the arachidonic acid (AA) cascade in the lungs of microwave exposed rats were investigated. One Sprague-Dawley male rats weighting $100{\pm}10$ g was randomly assigned to the normal group and three were assigned to the microwave exposed groups. The microwave exposed groups were subdivided into three groups according to the levels of dietary catechin supplementation: catechin free diet (MW) group, 0.25% catechin (MW-0.25C) group and 0.5% catechin (MW-0.5C) group. Rats were sacrificed on the 6th day after microwave irradiation (2.45 GHz, 15 min). The lung microsomal $PLA_2$ activity in the MW and MW-0.25C groups was 30% and 15% greater than that of the normal group, respectively, whereas no significant difference between the normal group and MW-0.5C group was observed. The percentage of phosphatidylethanolamine (PE) hydrolyzed in the lung microsome in the MW, MW-0.25C and MW-0.5C group increased by 47%, 18% and 20%, respectively, due to microwave irradiation. The formation of thromboxane $A_2$ ($TXA_2$) in the lung microsome was 50% greater in the MW group than in the normal group. However, the levels of $TXA_2$ in the MW-0.25C and MW-0.5C group were normal. The formation of prostacyclin ($PGI_2$) in the lung microsome was 31% lower in the MW group than in the normal group, while the levels of $PGI_2$ in the MW-0.25C and MW-0.5C group were similar to the normal group. The lung microsomal thiobarbituric acid reactive substances (TBARS) concentration, which can be used as an index of lipid peroxide was 34% greater in the MW group, when compared with the normal group. However, there was no difference between the MW-0.25C, MW-0.5C and normal groups. In conclusion, lung function appeared to be improved by green tea catechin supplementation due to its antithrombus action, which in turn controls the AA cascade system.

• Korean Journal of Pharmacognosy
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• v.52 no.3
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• pp.127-133
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• 2021

Platelet activation plays a major role in cardiovascular disorders (CVDs). Thus, disrupting platelet activation represents an attractive therapeutic target on CVDs. Esculetin, a bioactive 6,7-dihydroxy derivative of coumarin, possesses pharmacological activities against obesity, diabetes, renal failure, and CVDs. In other report, the effect of esculetin has been examined in human platelet activation and experimental mouse models, and esculetin inhibited collagen- and arachidonic acid-induced platelet aggregation in washed human platelets. However, it had no effects on other agonists such as thrombin and U46619, and its mechanism is not also clearly known. This study investigated the effect of esculetin on collagen-induced human platelet aggregation, and we clarified the mechanism. Esuletin has effects on the down regulation of PI3K/Akt and MAPK, phosphoproteins that act in the signaling process in platelet aggregation. The effects of esculetin reduced of TXA₂ production and phospholipase A₂ activation, and intracellular granule secretion including ATP and serotonin, leading to inhibit platelet aggregation. These results clearly clarified the effect of esculetin in inhibiting platelet activity and thrombus formation in humans.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.3
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• pp.418-427
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• 2003

Conjugated linoleic acid (CLA) is the mixture of positional and geometric isomers of linoleic acid (LA, C18:2 $\omega$6), which is found abundantly in dairy products and meats. This study was peformed to investigate the anticarcinogenic effect of CLA in MCF-7 breast cancer cells. MCF-7 cell were treated with LA and CLA at the various concentrations of 15, 30, 60, 120 UM each. After incubation for 48 and 72 hours, cell proliferation, fatty acids incorporation into cell, peroxidation and activities of antioxidant enzymes were measured. Postaglandin E$_2$ (PGE$_2$) and thromboxane $A_2$ (TXA$_2$) were measured for the eicosanoids metabolism. There was no cell growth differences in both of LA and CLA treated MCF-7 cells at 48 hr incubation. Compared to LA, cell growth was decreased by CLA treatment according to increasing concentration at longer incubation times, respectively (p<0.05). Both of LA and CLA was incorporated into the cellular lipids 22~54% higher than in control but LA incorporation was not so linear as CLA according to concentration. Arachidonic acid (C20:4, $\omega$6) was synthesized after treatment of LA but did not in CLA, respectively. The lipid peroxide concentration in LA 120 $\mu$M group increased as 1.7 times as that in CLA 120 $\mu$M treated. The activities of antioxidant enzymes such as glutathione peroxidase and glutathione reductase were increased by the supplementation with CLA 120 $\mu$M at 72 hr incubation (p<0.001) compared to LA, otherwise activity of superoxide dismutase was not different in both. PGE$_2$ and TXA$_2$ levels were lower in condition of CLA treatments according to lower levels of arachidonic acids than those in LA treated group, respectively. Overall, the dietary CLA might change the MCF-7 cell growth by the changes of cell composition, production of lipid peroxide, activities of antioxidant enzymes and eicosanoid synthesis compared to dietary LA.

• Journal of Applied Biological Chemistry
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• v.65 no.3
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• pp.167-172
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• 2022

Pathophysiological reaction of platelets in the blood vessel is an indispensable part of thrombosis and cardiovascular disease, which is the most common cause of death in the world. In this study, we performed in vitro assays to evaluate antiplatelet activity of artemether in human platelets and attempted to identify the mechanism responsible for protein phosphorylation. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artemether was clarified. Artemether inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artemether decreased TXA₂ production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artemether strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC₅₀ of 157.92 μM. These results suggest that artemether has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.122-127
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• 2023

Cardiovascular disease (CVD) is increasingly increasing as the main cause of death worldwide, and activation of platelet in vascular damage is one of the important causes of CVD. In recent, there is a growing interest in anti-thrombotic materials through platelet suppression, and efforts are being made to reduce side effects by using natural bioactive compounds. Known as one of the Flavonoids, hydroxygenkwanin (HGK) is a purified substance in Daphne Genkwa, which is known to have antibacterial, anti-inflammatory and anti-cancer effects, and has been reported to serve as an inhibitor of tissue factor that prevents thrombosis, but its anti-platelet effects and the action mechanisms is not known. In this study, we confirmed that the effects of HGK on the collagen-induced human platelets activation. HGK suppressed phosphorylation of PI3K/AKT and mitogen-activated protein kinases during platelet signaling, and reduced granule secretion in platelets such as ATP and serotonin. In addition, HGK inhibited the phosphorylation of cPLA₂ and strongly undermined the production of TXA₂, which is a powerful aggregation amplifier. As a result, the platelet aggregation derived by Collagen, a cohesive induced substance, was strongly suppressed by HGK to an IC₅₀ of 86.36 µM. Therefore, HGK might be worth the antithrombotic substance that inhibits the activation and aggregation of human platelets that occur through blood vessel damage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.175-181
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• 1998

The relation between lipid peroxidation and thrombotic reaction were investigated in streptozotocin (STZ)-induced diabetic rats. Sprague-Dawley male rats weighing 100$\pm$10gm were randomly assigned to normal and STZ-induced diabetic group(DM). Diabetes was experimentally induced by intravenous injection of 55mg/kg of body weight of STZ in citrate buffer(pH 4.3) after 4 weeks feeding of basal diet. Animals were sacrificed at the 6th day of diabetic states. Body weight gains were lower in diabetic group after STZ injection. Serum levels of thiobarbituric acid reacting substances(TBARS) that were markedly increased in DM group compared with of normal group. TBARS levels of HDL and LDL were similar patterns to total TBARA of serum. Activities of platelet phospholipase A2(PLA2) were higher in diabetic group than those of normal group. Activities of platelet cyclooxygenase were 106% in DM group than normal group. Platelet thromboxane A2(TXA2) formation was increased in DM group than normal group. Production of aortic prostacyclin(PGI2) was lower in diabetic group than that of normal group. PGI2/TXA2 ratios were decreased by 55% in DM groups than those of normal group. The present results indicate that STZ-induced diabetic rats are more sensitive to oxidative stess which leads to acceleration of lipid peroxidation and platelet aggregability. In conclusion, accelerating effect of lipid peroxidation and thrombogenesis in diabetic state is regareded to be resulted from enhancement of PLA2 activity and arachidonic acid metabolism, inhibition of antiaggrgating agent and aortic PGI2 formation.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.5
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• pp.463-468
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• 2013

Acute hypoxia induces contraction of pulmonary artery (PA) to protect ventilation/perfusion mismatch in lungs. As for the cellular mechanism of hypoxic pulmonary vasoconstriction (HPV), hypoxic inhibition of voltage-gated $K^+$ channel (Kv) in PA smooth muscle cell (PASMC) has been suggested. In addition, our recent study showed that thromboxane $A_2$ ($TXA_2$) and hypoxia-activated nonselective cation channel ($I_{NSC}$) is also essential for HPV. However, it is not well understood whether HPV is maintained in the animals exposed to ambient hypoxia for two days (2d-H). Specifically, the associated electrophysiological changes in PASMCs have not been studied. Here we investigate the effects of 2d-H on HPV in isolated ventilated/perfused lungs (V/P lungs) from rats. HPV was almost abolished without structural remodeling of PA in 2d-H rats, and the lost HPV was not recovered by Kv inhibitor, 4-aminopyridine. Patch clamp study showed that the hypoxic inhibition of Kv current in PASMC was similar between 2d-H and control. In contrast, hypoxia and $TXA_2$-activated $I_{NSC}$ was not observed in PASMCs of 2d-H. From above results, it is suggested that the decreased $I_{NSC}$ might be the primary functional cause of HPV disappearance in the relatively early period (2 d) of hypoxia.

• Journal of Ginseng Research
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• v.21 no.2
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• pp.132-140
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• 1997

The effects of ginsenosides purified from red ginseng on platelet aggregation were investigated. Preincubation of washed platelets from rats with either ginsenoside Rg3, ginsenosides non-polar fraction (G-NPF), ginsenoside Rg1(Rg1) or ginsenosides polar fraction(G-PF) reduced the plytelet aggrelation induced by collagen in a dose-dependent manner, whereas ginsenoside Rg2 failed to inhibit the aggregation. Their IC50 values of Rg3, G-NPF, Rgl, and G-PF were 8.7$\pm$1.0, 150.3$\pm$0.1, 369.9$\pm$ 1.0, 606.211.3 $\mu\textrm{g}$/ml, respectively. Aggrelation induced by thrombin was also inhibited by Rg3 and G-NPF with IC50 being 5.2$\pm$ 1.1 and 66.5$\pm$0.8 $\mu\textrm{g}$/ml, respectively. The alterations of Intracellular Ca2+ concentration in platelets were monitored using fura-2 as a fluorescent Ca2+ indicator. Both Ca2+ release from internal stores and Ca2+ influx into cytosol were suppressed by Rg3. Rg3 also inhibited granular release of ATP and TXA2 formation induced by thrombin in a dose-dependent manner in the washed platelets. Rg3 also inhibited Aggregation and ATP release from human platelets induced by collagen to a similar extent as were observed in rat platelets. In conclusion, Rg3 is a Potent anti-aggregating component in ginsenosides and may exert its anti-aggrega1ing activity by decreasing TXAa formation and granular secretion in platelets, most likely by inhibiting Ca2+ influx and Ca2+ mobilization from intracellular stores. Thus ginseng may contribute to the prevention and treatment of thrombosis.

• Journal of Nutrition and Health
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• v.28 no.2
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• pp.95-106
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• 1995

The effects of various n-6/n-3 ratios(about 2, 4, 6, 8) of dietary fatty acids on various lipid levels and prostaglandin production were studied at the constant P/S ratio (1.5-1.6) in young (5 weeks old) and adult(8 months old) Sprague-Dawley rats using palm oil, safflower oil and sardine oil. The concentration of serum cholesterol tended to increase with the increasing n-6/n-3 ratio. The tendency of HDL-cholesterol levels was similar to serum cholesterol levels. These were not apparent differences between young and adults rats. Serum triglyceride levels increased according to increasing n-6/n-3 ratio in young rats. These were generally high in the adult rats compared with the young rats. Though liver cholesterol level tended to increase according to the increasing n-6/n-3 ratio in the young rats. The liver triglyceride level did not change according to the n-6/n-3 ratio. However, these levels were apparently higher in the adult than in the young rats. The fatty acid compositions of phosphatidylcholine(PC) were similar in serum and liver. The arachidonate/linoleate ratios in tissue PC were influenced by the n-6/n-3 ratio. They tended to be lower in the adult rats compared with the young rats. It was suggested that the activity of $\Delta$6-desturase was decreased by aging. Production of platelet thromboxane A2(TXA2)and aortic prostacyclin(PGI2) was not apparently influenced with n-6/n-3 ratio. Whereas the ratio of TXA2/PGI2 was the lowest value at 3.8 of n-6/n-3 ratio, expecially in the young rats. Thus this ratio seemed to be a desirable level to protect atherosclerosis. These results indicate that the lipid level and prostaglandin production were influenced not only by n-6/n-3 ratio(under constant P/S ratio) but by aging, particulary triglycerde level and arachidonic/linoleic acid ratio.

• KSBB Journal
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• v.28 no.1
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• pp.7-12
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• 2013

Thromboxane $A_2$ ($TXA_2$) is one of major proinflammatory mediators, plays an important role in the development of vascular inflammatory diseases. $TXA_2$ acting through the thromboxane receptor regulates multiple pathways and genes in a variety of cells. In this study, we report that the activation of thromboxane receptor with U46619 increases the interleukin-8 (IL-8) mRNA in vascular endothelial cells. We also demonstrated that U46619 produces the activations of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (MAPK), which is required for endothelial IL-8 production. And U46619 enhanced mRNA stability of IL-8 transcripts in endothelial cells. Moreover, inhibition of ERK1/2 or p38MAPK reduced monocyte adhesion to aortic endothelium stimulated by U46619. Therefore, these results suggest that activation of thromboxane receptor promotes the expression of IL-8 via ERK1/2 and p38MAPK activation in endothelial cells.