• Journal of Food Hygiene and Safety
• /
• v.10 no.2
• /
• pp.73-79
• /
• 1995

Incidents of seafood and massive fish kills have been rapidly increasing in both frequency and geographical distribution and the socioeconomic impacts brought by those incidents. However, the biological origins of those marine toxins have not been well clarified. Most of the marine organisms investigated are filter-feeder, which accumulate toxins from their food and/or their symbiotic microalgae. We have examined the action on rabbit platelets of marine toxins isolated from cultured dinoflagellates and sponge collected at Okinawa. Maitotoxin (MTX) is a water-soluble toxin isolated from the cultured dinoflagellate Gambierdiscus toxicus which causes a seafood poisoning in tropical regions. Zooxanthellatoxin A (ZT-A) was isolated from exteracts of cultured symbiotic dinoflagellate Symbiodinium sp. (socalled zooxanthella) from flatworms of the genus Amphiscolops collected at Okinawan marine sponge Theonella sp. MTX caused a disaggregation and a dissolution of large aggregates. ZT-A caused a dissolution of small aggregates followed by a increment of light tranmission. TZ-A caused an initial and transient shape change followed by a sustained aggregation and a increment of large aggregates. In conclusion, marine toxins exert unique patterns on the light trasmission and the size of aggregates in rabbit platelets by their concentrations and kinds

• The Korean Journal of Physiology
• /
• v.29 no.2
• /
• pp.217-223
• /
• 1995

Adenosine 5'-tetraphosphate (ATPP) was identified and quantified in extracts of rabbit platelets by elution of extracts containing authentic adenosine 5'-tetraphosphate and comparison of retention time with nucleotide standards using high-performance liquid chromatography technique. The amount of adenosine 5'-tetraphosphate was $0.62\;nmoles/10^{9}$ cells which was 62-fold lower than that of ATP but only 10-fold lower than that of ADP. During platelet aggregation induced by thrombin, adenosine 5'-tetraphosphate was released to a relatively high extent. The degradation rates and halflives of adenosine 5'-tetraphosphate were measured during incubation of platelets in whole blood, erythrocyte suspension and plasma, respectively. The results suggest that plasma contributes more than blood cells to the catabolism of adenosine 5'-tetraphosphate. The pattern of degradation indicates that ATPP may be degraded mainly to AMP by soluble enzymes in plasma and very slowly to ADP and/or AMP by ectoenzymes on blood cells such as erythrocyte. The nature of the enzymes responsible fer the degradation of adenosine 5'-tetraphosphate is yet to be identified.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.3
• /
• pp.511-515
• /
• 2002

Paeoniae radix has been considered as one of the most important crude drugs used in traditional oriental medicine and has been employed as a circulatory tonic in care of weakness, night sweats, and lumbar pain, etc. Platelet activation plays an important role in thrombosis and haemostasis. Active compounds for the inhibition of platelet activation from Paeoniae radix were extracted and fractionated into five fractions. Its fraction two and three of ethyl acetate extract inhibited the aggregation of washed rabbit platelets induced by collagen. Two active compounds, bezoyloxypaeoniflorin and paeoniflorin, were isolated from fraction two and three by silica gel column and high performance liquid chromatography. The chemical structures were determined by comparison of their proton and carbon nuclear magnetic resonance spectra. Benzoyloxypaeoniflorin showed strong inhibition at the concentration of 100 ug/mL against collagen-induced washed rabbit platelets aggregation. It is suggested that Paeoniae radix has become food material to prevent a cardiovascular disease.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.110.3-111
• /
• 2003

In the previous studies, we have reported that NQ30l, a synthetic l,4-naphthoquinone derivative, displayed a potent antithrombotic activity, and that this might be due to antiplatelet effect, which was mediated by inhibition of cytosolic $Ca^{2+}$ mobilization in activated platelets. In the present study, the effect of NQ301 on arachidonic acid cascade in activated platlets was examined. NQ301 concentration-dependently inhibited washed rabbit platelet aggregation induced by collagen (10 $\mu$g/ml), arachidonic acid (100 $\mu$M) and U46619 (1 $\mu$M), a thromboxane $A_2$receptor agonist, with $IC_50$ values of 0.60$\pm$0.02, 0.79$\pm$0.04 and 0.58$\pm$0.04 $\mu$M, respectively. (omitted)

• Proceedings of the KAIS Fall Conference
• /
• 2004.06a
• /
• pp.253-257
• /
• 2004

Platelet aggregation is a complex phenomenon that probably involves several intracellular biochemical pathways. When activated, platelets change shape, aggregate and release the contents of their intracellular granules. The interactions between platelets and blood vessel walls are important in the development of thrombosis and cardiovascular diseases. When blood vessels are damaged, platelet aggregation occurs rapidly to form haemostatic Plugs or arterial thrombi at the sites of vessel injury or in regions where blood flow is disturbed. These thrombi are the source of thromboembolic complications of atherosclerosis, heart attacks, stroke, and peripheral vascular disease. Therefore, the inhibition of platelet function represents a promising approach for the prevention of thrombosis. Plants constitute a rich source of bioactive chemicals such as phenolics, terpenoids and alkaloids. Plant extracts may be an alternative to currently used medicinal source because they constitute a rich source of bioactive chemicals. This study was performed to investigate the antiplatelet activity of extract of Tabebuia impetiginosa Martius ex DC (Taheebo) and find out which fractions to this activity in rabbit platelet. Taheebo was methanol extracted and solvent fractionated in to five fractions (hexane, chloroform, ethylacetate, butanol and water). And each fractions were investigated inhibitory effects on platelet aggregation induced by various agonists using washed rabbit platelets in vitro.

• Journal of Food Hygiene and Safety
• /
• v.15 no.2
• /
• pp.167-172
• /
• 2000

Lithospermum erythrorhyzon has been used as a red fooddye and traditional Chinese medicine to treat wounds, skin diseases and burns. Platelet activation plays an important role in thrombosis and haemostasis. Here, we studied the inhibition of platelet activation and its active compound from the root of Lithospermum erythrorhyzon. Its ethyl acetate extract inhibited the aggregation of washed rabbit platelets induced by collagen or thrombin. Five naphthoquinone pigments , shikonin , acetylshikonin , is obutylshikonin, $\alpha$-methyl-n-butylshikonin and $\beta$,$\beta$-dimethylacrylshikonin were isolated by means of high pressure liquid chromatography. The structures were determined by comparison of their proton nuclear magnetic resonance spectra. The potency of their inhibition was in the following order : $\beta$,$\beta$-dimethylacrylshikonin$\geq$$\alpha$-methyl-n-butylshikonin>isobutylshikonin>acetylshikonin>shikonin. It is suggested that the size of the aliphatic hydroxy group of shikonin is important for the enhancement of potency.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.78.2-78.2
• /
• 2003

We have previously reported that green tea catechins (GTC) displayed anti-thrombotic activity, and that this might be due to anti-platelet rather than anti-coagulation effects. In the present study, we have studied the anti-platelet activity and mechanism of gallocatechin gallate (GCG), which is a component of GTC. GCG inhibited the collagen- and U46619-induced aggregation of rabbit platelets, with IC$\^$50/ values of 63.0 and 48.3 ${\mu}$M, respectively. GCG also inhibited collagen-induced serotonin release and TxB$_2$ formation in a similar manner of platelets aggregation. (omitted)

• Archives of Pharmacal Research
• /
• v.29 no.5
• /
• pp.375-383
• /
• 2006

The anti platelet effects of a novel guanidine derivative, KR-32570 ([5-(2-methoxy-5-chlorophenyl) furan-2-ylcarbonyl]guanidine), were investigated with an emphasis on the mechanisms underlying its inhibition of collagen-induced platelet aggregation. KR-32570 significantly inhibited the aggregation of washed rabbit platelets induced by collagen $(10{\mu}g/mL)$, thrombin (0.05 U/mL), arachidonic acid $(100{\mu}M)$, a thromboxane (TX) $A_2$ mimetic agent U46619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin $F_2,\;1{\mu}M$) and a $Ca^{2+}$ ATPase inhibitor thapsigargin $(0.5{\mu}M)$ ($IC_{50}$ values: $13.8{\pm}1.8,\;26.3{\pm}1.2,\;8.5{\pm}0.9,\;4.3{\pm}1.7\;and\;49.8{\pm}1.4{\mu}M$, respectively). KR-32570 inhibited the collagen-induced liberation of $[^3H]$arachidonic acid from the platelets in a concentration dependent manner with complete inhibition being observed at $50{\mu}M$. The $TXA_2$ synthase assay showed that KR-32570 also inhibited the conversion of the substrate $PGH_2$ to $TXB_2$ at all concentrations. Furthermore, KR-32570 significantly inhibited the $[Ca^{2+}]_i$ mobilization induced by collagen at $50{\mu}M$, which is the concentration that completely inhibits platelet aggregation. KR-32570 also decreased the level of collagen $(10{\mu}g/mL)$induced secretion of serotonin from the dense-granule contents of platelets, and inhibited the NHE-1-mediated rabbit platelet swelling induced by intracellular acidification. These results suggest that the antiplatelet activity of KR-32570 against collagen-induced platelet aggregation is mediated mainly by inhibiting the release of arachidonic acid, $TXA_2$ synthase, the mobilization of cytosolic $Ca^{2+}$ and NHE-1.

• Archives of Pharmacal Research
• /
• v.25 no.6
• /
• pp.873-878
• /
• 2002

OA-8159, a new Phosphodiesterase (PDE) 5 inhibitor, has exhibited potent erectogenic potential in a penile erection test in rats and anesthetized dogs. In this study, we investigated the mechanism of its erectogenic activity by measuring the activity of OA-8159 against a various PDE isozymes and assessing cGMP and cAMP formation in a rabbit corpus cavernosum in vitro. DA-8159 inhibited the PDE 5 activity in rabbit and human platelets, which the $IC_{50}$ was 5.84$\pm$1.70 nM and 8.25$\pm$2.90 nM, respectively. The $IC_{50}$ of DA-8159 on PDE 1, PDE2, PDE 3 and PDE 6 were 870$\pm$57.4 nM, $101\pm$5 $\mu$M, 52.0$\pm$3.53 $\mu$M and 53.3$\pm$2.47 nM, respectively. This suggests that DA-8159 is a potent, highly selective, competitive inhibitor of PDE 5-catalyzed cGMP hydrolysis. The rates of cGMP hydrolysis catalyzed by human platelets-derived PDE 5 as a function of the cGMP concentration (5~100 nM) and two-fixed DA-8159 concentration (11.3 and 18.8 nM) were investigated in order to characterize the mode of PDE 5 inhibition by DA-8159. DA-8159 increased the apparent 4K_{m}$ value for cGMP hydrolysis but had no effect on the apparent $V_{max}$, indicating a competitive mode of inhibition. DA-8159 increased the cGMP concentrations in the rabbit corpus cavernosum dose dependently. In the presence of sodium nitroprusside (SNP), DA-8159 significantly sti\mulated the accu\mulation of cGMP when compared to the control level. This indicated that the enhancement of a penile erection by DA-8159 involved the relaxation of the cavernosal smooth \muscle by NO-sti\mulated cGMP accu\mulation. In conclusion, DA-8159 is a selective inhibitor of PDE 5-catalyzed cGMP hydrolysis and the enhancement of a penile erection by DA-8159 is mediated by the relaxation of the cavernosal smooth \muscle by the NO-sti\mulated cGMP accu\mulation.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1996.04a
• /
• pp.203-203
• /
• 1996

It has been known that pinusolide isolated from Biota orientalis inhibits specifically [$^3$H] PAF binding to and PAF-induced aggregation of rabbit platelets in vitro. The present study was undertaken to evaluate the effects of pinusolide on hypotension and bronchoconstriction due to PAF. PAF(i.v.) in doses of 0.01 to 0.3 $\mu\textrm{g}$/kg induced a dose-dependent hypotension without tachyphylaxis in rat. Pinusolide(5～20 mg/kg, i.v.) inhibited PAF(0.03 $\mu\textrm{g}$/kg, i.v.)-induced hypotension dose-dependently, while it failed to block the hypotensive action of histamine, serotonin and acetylcholine. It inhibited bronchoconstriction and cutaneous vascular permeability induced by PAF in anesthetized guinea-pigs and rats, respectively, but it showed no inhibitory effect on the increase in bronchial resistance by histamine and acetylcholine. These results strongly suggest that in vivo pinusolide acts as a selective antagonist of PAF.