桂枝茯笭丸 및 그 構成藥物의 血小板凝集抑制에 關한 硏究

金宗求,朴宣東,朴元煥 동국대학교 한의학연구소 2000 東國韓醫學硏究所論文集 Vol.8 No.2

動物性 脂肪攝取量의 增加, 運動不足, 肥滿, 스트레스의 加重, 高齡化의 增加 等의 原因으로 循環器系疾患의 發病率이 增加하고 있으며, 이러한 循環器系疾患의 危險因子로서 血栓症이 重要하게 대두되고 있다. 특히 최근 문제시되고 있는 狹心症이나 心筋梗塞등의 虛血性 心疾患은 血小板凝集에 의해 일어나는 血栓形成에 起因하고 있다. 韓醫學에서 血栓症은 瘀血의 範疇에 屬하며, 瘀血은 각종 病理的 原因에 의해 발생한 全身性 또는 局所性의 血液循環 障碍 또는 血流停滯와 그에 수반되는 일련의 病症을 나타내며, 驚悸정충, 鼓脹, 積聚, 미하, 癲狂, 中風等의 發病原因이 된다. 또한 瘀血에 의한 각종 症候에는 活血祛瘀劑 또는 驅瘀血劑등이 사용되고 있다. 本 硏究에서는 韓醫學에서 瘀血症으로 惹起되는 여러 가지 症狀의 개선에 사용되는 驅瘀血劑들의 血小板凝集에 미치는 영향을 검색하기 위하여 桂枝茯笭丸(Geijibokryunghwan; GBH) 및 그 構成藥物을 使用하였다. 桂枝茯笭丸은 『금궤요략』에 있는 方으로써 祛邪不傷正하고 調氣寒熱하여 예로부터 驅瘀血劑로 사용되어 왔다. 이에 桂枝茯笭丸 및 그 構成藥劑의 ADP, AA 또는 collagen으로 유도되는 血小板凝集에 대하여 抑制效果를 探索한 結果, 桂枝茯笭丸 및 個別 構成藥物의 血小板凝集抑制作用을 확인하였고, 血小板凝集으로 惹起되는 血栓症등에 桂枝茯笭丸 및 個別 構成藥物은 매우 臨床實驗的 應用價値가 있는 것으로 생각되었다. The cause that the increase of animality fat intakes, under exercise, fatness, adding the stress, advanced age etc., the occurrence rate of the circulation system &sea= has been increased. And the thrombosis importantly came to the front as the risk factor of these circulation system's disease. Nowadays, the ischemic disease has especially discussed, for example the angina or myocardial infarction, originated in thrombosis that came from the platelet aggregation. In the western medicine, as the cure and prevention, using the aspirin or ticlopidine for platelet aggregation suppressant. But in the <Synopsis of the golden chamber : On Pulse, Syndrome Complex and Treatment of Chest Obstruction, Heart Pain and Shortness of Breath>, the curing method must be used properly according to the pectoralgia or heartache's lund, state, grade. The platelet do not attache to the normal hemangioendothelial cell. But when It stimulated by endothelium peronia and so on, it attache to the injury endothelium or rise aggregation between the platelet. On this time, it secrete the platelet aggregation inducer as like ADP, thromboxane A2 from the inside of platelet. So it has first defensive function through the aggregation augment that prevent the celerity consumption of blood. But the activation of abnormal platelet occur the platelet grume and thrombogenesis. So it bring up the occlusive angiosis. so to speak, cardiovascular disease, cerebrovascular disease, arterial sclerosis. In oriental medicine, the thrombosis in the category of blood stasis and this blood stasis present the generalise or local blood circulation disturbance that generated by all kinds of pathological fact or blood stream retention accompanying with a series of syndrome. As the syndrome, stabbing pain fixed at certain region, squamous and dry skin, fullness and pain of the chest and hypochondrium, firmness and fullness of the lower abdomen, black stool, dark purple tongue or with ecchymoses and petechiae etc.. has been created. And it becomes the pathopoiesis cause that the convulsion and palpitation, severe palpitation. tympanites, the symtom complex with a mass or swelling in the abdomen, insanity, stricken by wind etc.. Moreover, it used the drugs for invigorating blood circulation and eliminating blood stasis or drugs for removing blood stasis for all kinds of syndrome through the blood stasis. And the drugs for activating the blood circulation, such as Salviae Radix, Angelicae Sinensis Radix, Persicae Semen, Achyranthis Radx, Cnidii Rhizoma, Carthami Flos are used for that. And it is used to the herbs of insects that has strong effect about the disintergrating blood stasis such as Hirudo, Scolopendrae Corpus, Buthus, Lumbricus etc.. On this study, It used Geijibokryunghwan(GBH) and the consisting herbs to investigate the influence of platelet aggregation about drugs that used to improvement various symptoms created by the thrombosis in oriental medicine. GBH formula has as formula recorded in the <Synopsis of the golden chamber>, action of 'eleminating the evil and not impairment of healthy energy' and 'promoting the flow of QI and cold and heat, so used for the expel blood stasis herbs from the ancient. Therefore we investigated the restraint effect of GBH and the consisting herbs about the platelet agregation induced to the ADP, AA or collagen. The conclusion is following. 1. When it added the aggregation inducer after that it added GBH and individual consisting herbs in the PRP, GBH showed the (+) inhibition effect on the platelet aggregation and it showed the (+) inhibition effect in the individual consisting herbs as like Paeoniae Radix and Moutan Cortex Radicis. 2. It showed the (+), (+,++) inhibition effect on the platelet aggregation in Paeoniae Radix·Hoelen, Paeoniae Radix· Moutan Cortex Radicis, Hoelen· Moutan Cortex Radicis etc. 3. In the aggregation inhibition activating on the difference of density, GBH showed strong ihbition effect to the aggregation state induced to collagen, and it showed the inhibition effect in the individual consisting herbs as like Paeoniae Radix and Moutan Cortex Radicis about the aggregation induced by the collagen. 4. It showed the strong inhibition effect about the aggregation induced by the collagen in Paeoniae Radix·Hoelen, Paeoniae Radix· Moutan Cortex Radicis, Hoelen·Moutan Cortex Radicis etc Like this, as confirm GBH and the individual consisting herb's inhibition effect of platelet aggregation, We considerated that GBH and the individual consisting herbs have practical applicational value of clinical trial in the thrombosis caused by platelet aggregation.

인슐린비의존형 당뇨병 환자에서 단시간 운동부하에 의한 혈소판응집능의 감소

정동진(Dong Jin Chung),장현주(Hyun Ju Jang),안재수(Jae Su An),최종상(Jong Sang Choi),윤재영(Jae Young Yoon),정민영(Min Young Chung),이태희(Tai Hee Lee) 대한내과학회 1991 대한내과학회지 Vol.41 No.2

N/A Increased platelet aggregation in diabetes mellitus is considered to be one of the etiologic factors of diabetic microangiopathy. The mechanism influencing platelet aggregation by physical exercise in non-insulin-dependent diabetes mellitus (NIDDM) is poorly understood. This study was performed to evaluate the effectiveness of short-term exercise on platelet aggregation in patients with non-obese, non-insulin-dependent diabetes mellitus compared with the normal group. Before and after submaximal exercise using ergometer, platelet aggregation responses to adenosine diphosphate (ADP), collagen, and epinephrine were measured, and the following results were obtained. 1) In platelet aggregation test, in NIDDM patients (n=45), the maximal platelet aggregations [to ADP; 78.7% (p<0.01), to epinephrine; 74.5% (p<0.05)] were significantly higher than those [to ADP; 65.1%, to epinephrine; 67.3%] of control subjects(n=15). 2) In control subjects (n =9), platelet aggregations after exercise was not significantly changed [before exercise; ADP 65.8±3.55%, collagen 70.2±2.35%, epinephrine 58.3±6.08%, after exercise; ADP 68.0±2.84%, collagen 68.9±2.46%, epinephrine 59.5±5.73%, respectively]. 3) In NIDDM patients (n =13), platelet aggregations to ADP and collagen after exercise were significantly decreased [before exercise; ADP 73.4±3.98%, collagen 79.3±4.24%, epinephrine 72.8±3.57%, after exercise; ADP 67.9±3.27% (p<0.05), collagen 69.3±3.49% (p<0.01), epinephrine 66.8±3.97%, respectively]. 4) There were no differences in the plasma thromboxane B2 levels between controls (4.6±1.44 pg/ml) and NIDDM patients (8.5+1.95pg/ml), and after exercise, thromboxane B2 was decreased, but not significantly, in NIDDM patients (before exercise; 18.1±5.36pg/ml, after exercise; 11.5±2.58 pg/ml). 5) Duration of diabetes, hemoglobin Ai, and fasting blood sugar did not affect the platelet aggregation. These results indicate that short-term exercise decreases platelet aggregation in NIDDM patients.

허혈성 뇌졸중 환자에서 신속혈소판기능 검사를 이용한 클로피도그렐의 혈소판 응집 억제 반응 측정

김재국,김형일,신혜은,이보람,전종은,이수주 대한신경과학회 2008 대한신경과학회지 Vol.26 No.4

Background: Clopidogrel inhibits platelet P2Y12 adenosine diphosphate (ADP) receptors and has been widely used in patients with ischemic stroke. However, a considerable number of patients suffer from cerebrovascular events despite the use of clopidogrel. The rapid platelet function assay (RPFA) has been used for monitoring the antiplatelet effects on the P2Y12 ADP receptor. This study was performed to measure the platelet response to clopidogrel using RPFA in patients with ischemic stroke, and to identify the clinical factor related with clopidogrel resistance. Methods: A total of 86 patients taking clopidogrel (75 mg/day) were enrolled. Demographic data, vascular risk factors, the presence of obesity and metabolic syndrome, drug history, hemoglobin, platelet counts, and stroke subtypes were recorded. RPFA presented the results as P2Y12 Reaction Units (PRU), base PRU (BASE), and Inhibition (%). Inhibition was calculated as (1-PRU/BASE)×100. The patients showing ineffective aggregation- inhibition (percentage of Inhibition < 20) on RPFA were defined as non-responders to clopidogrel. Results: The response of platelet aggregation-inhibition to clopidogrel showed a variable distribution with mean and standard deviation of 32.2±22.3%. Twenty four (27.9%) patients showed the inhibition below 20%. There was no difference between responders and non-responders regarding the clinical factors above. We found no influence of co-medication with the statins on platelet response to clopidogrel. Conclusions: There is a patient variability in response to clopidogrel and a considerable portion of stroke patients have clopidogrel resistance on the platelet function test. The clinical usefulness of routine platelet function test requires further validation. Background: Clopidogrel inhibits platelet P2Y12 adenosine diphosphate (ADP) receptors and has been widely used in patients with ischemic stroke. However, a considerable number of patients suffer from cerebrovascular events despite the use of clopidogrel. The rapid platelet function assay (RPFA) has been used for monitoring the antiplatelet effects on the P2Y12 ADP receptor. This study was performed to measure the platelet response to clopidogrel using RPFA in patients with ischemic stroke, and to identify the clinical factor related with clopidogrel resistance. Methods: A total of 86 patients taking clopidogrel (75 mg/day) were enrolled. Demographic data, vascular risk factors, the presence of obesity and metabolic syndrome, drug history, hemoglobin, platelet counts, and stroke subtypes were recorded. RPFA presented the results as P2Y12 Reaction Units (PRU), base PRU (BASE), and Inhibition (%). Inhibition was calculated as (1-PRU/BASE)×100. The patients showing ineffective aggregation- inhibition (percentage of Inhibition < 20) on RPFA were defined as non-responders to clopidogrel. Results: The response of platelet aggregation-inhibition to clopidogrel showed a variable distribution with mean and standard deviation of 32.2±22.3%. Twenty four (27.9%) patients showed the inhibition below 20%. There was no difference between responders and non-responders regarding the clinical factors above. We found no influence of co-medication with the statins on platelet response to clopidogrel. Conclusions: There is a patient variability in response to clopidogrel and a considerable portion of stroke patients have clopidogrel resistance on the platelet function test. The clinical usefulness of routine platelet function test requires further validation.

Inhibitory Effect of Cordycepin on Human Platelet Aggregation

박화진,--,--,--,--,--,-- THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2004 Journal of biomedical laboratory sciences Vol.10 No.1

Cordycepin separated from Cordyceps militaris is a major physiologic active component in Cordyceps militaris. The platelet aggregation is stimulated by Ca^(2+), which is either mobilized from intracellular endoplasmic reticulum or transported from extracellular space. cGMP antagonizes the actions of Ca^(2+). Based on these facts, we have investigated the effects of cordycepin on the mobilization of Ca^(2+) and the production of cGMP on collagen (10 ㎍/ml)-induced human platelet aggregation. Cordycepin potently stimulated the human platelet aggregation induced by collagen (10 ㎍/ml) in a dose-dependent manner. Cordycepin (500 μM) inhibited also the collagen-induced human platelet aggregation in the presence both 1 mM and 2 mM of CaCl_(2). These are in accord with the results that cordycepin inhibited the Ca^(2+)-influx on collagen-induced human platelet aggregation. These results suggest that cordycepin decrease the intracellular Ca^(2+) concentration to inhibit collagen-induced human platelet aggregation. Besides, cordycepin increased the level of cGMP on collagen-induced human platelet aggregation. This result is related with the decrease of intracellular Ca^(2+) concentration, because cGMP inhibits the mobilization of Ca^(2+). In addition, cordycepin inhibited the human platelet aggregation induced by LY-83583, inhibitor of guanylate cyclase. This result suggested that cordycepin inhibit the platelet aggregation by stimulating the activity of guanylate cyclase. In conclusion, we demonstrated that cordycepin might have the antiplatelet function by inhibiting Ca^(2+)-mobilization via the stimulation of the production of cGMP.

Inhibitory Effect of Cordycepin on Human Platelet Aggregation

Cho, Hyun-Jeong,Ham, Hye-Seon,Lee, Tae-Kyung,Jung, Young-Jin,Park, Sun-A,Kang, Hyo-Chan,Park, Hwa-Jin The Korean Society for Biomedical Laboratory Scien 2004 Journal of biomedical laboratory sciences Vol.10 No.1

Cordycepin separated from Cordyceps militaris is a major physiologic active component in Cordyceps militaris. The platelet aggregation is stimulated by $Ca^{2+}$, which is either mobilized from intracellular endoplasmic reticulum or transported from extracellular space. cGMP antagonizes the actions of $Ca^{2+}$. Based on these facts, we have investigated the effects of cordycepin on the mobilization of $Ca^{2+}$ and the production of cGMP on collagen ($10\mu$g/ml)-induced human platelet aggregation. Cordycepin potently stimulated the human platelet aggregation induced by collagen ($10\mu$g/ml) in a dose-dependent manner. Cordycepin (500 $\mu$M) inhibited also the collagen-induced human platelet aggregation in the presence both 1 mM and 2 mM of $CaCl_2$. These are in accord with the results that cordycepin inhibited the $Ca^{2+}$- influx on collagen-induced human platelet aggregation. These results suggest that cordycepin decrease the intracellular $Ca^{2+}$ concentration to inhibit collagen-induced human platelet aggregation. Besides, cordycepin increased the level of cGMP on collagen-induced human platelet aggregation. This result is related with the decrease of intracellular $Ca^{2+}$ concentration, because cGMP inhibits the mobilization of $Ca^{2+}$. In addition, cordycepin inhibited the human platelet aggregation induced by LY -83583, inhibitor of guanylate cyclase. This result suggested that cordycepin inhibit the platelet aggregation by stimulating the activity of guanylate cyclase. In conclusion, we demonstrated that cordycepin might have the antiplatelet function by inhibiting $Ca^{2+}$-mobilization via the stimulation of the production of cGMP.

Inhibitory Effect of Cordycepin on Human Platelet Aggregation

Hyun-Jeong Cho,Hye-Seon Ham,Tae-Kyung Lee,Young-Jin Jung,Sun-A Choi,Hyo-Chan Kang,Hwa-Jin Park 대한의생명과학회 2004 Biomedical Science Letters Vol.10 No.1

Cordycepin separated from Cordyceps militaris is a major physiologic active component in Cordyceps militaris. The platelet aggregation is stimulated by Ca²?, which is either mobilized from intracellular endoplasmic reticulum or transported from extracellular space. cGMP antagonizes the actions of Ca²?. Based on these facts, we have investigated the effects of cordycepin on the mobilization of Ca²? and the production of cGMP on collagen (10 ㎍/㎖)-induced human platelet aggregation. Cordycepin potently stimulated the human platelet aggregation induced by collagen (10 ㎍/㎖) in a dose-dependent manner. Cordycepin (500 μM) inhibited also the collagen-induced human platelet aggregation in the presence both 1 mM and 2 mM of CaCl₂. These are in accord with the results that cordycepin inhibited the Ca²?-influx on collagen-induced human platelet aggregation. These results suggest that cordycepin decrease the intracellular Ca²? concentration to inhibit collagen-induced human platelet aggregation. Besides, cordycepin increased the level of cGMP on collagen-induced human platelet aggregation. This result is related with the decrease of intracellular Ca²? concentration, because cGMP inhibits the mobilization of Ca²?. In addition, cordycepin inhibited the human platelet aggregation induced by LY-83583, inhibitor of guanylate cyclase. This result suggested that cordycepin inhibit the platelet aggregation by stimulating the activity of guanylate cyclase. In conclusion, we demonstrated that cordycepin might have the antiplatelet function by inhibiting Ca²?-mobilization via the stimulation of the production of cGMP.

The anti-platelet activity of panaxadiol fraction and panaxatriol fraction of Korean Red Ginseng in vitro and ex vivo

Yuan Yee Lee,Yein Oh,Min-Soo Seo,Min-Goo Seo,Jee Eun Han,Kyoo-Tae Kim,Jin-Kyu Park,Sung Dae Kim,Sang-Joon Park,Dongmi Kwak,Man Hee Rhee 고려인삼학회 2023 Journal of Ginseng Research Vol.47 No.5

Background: The anti-platelet activity of the saponin fraction of Korean Red Ginseng has been widelystudied. The saponin fraction consists of the panaxadiol fraction (PDF) and panaxatriol fraction (PTF);however, their anti-platelet activity is yet to be compared. Our study aimed to investigate the potency ofanti-platelet activity of PDF and PTF and to elucidate how well they retain their anti-platelet activity viadifferent administration routes. Methods: For ex vivo studies, Sprague-Dawley rats were orally administered 250 mg/kg PDF and PTF for 7consecutive days before blood collection via cardiac puncture. Platelet aggregation was conducted afterisolation of the washed platelets. For in vitro studies, washed platelets were obtained from Sprague-Dawley rats. Collagen and adenosine diphosphate (ADP) were used to induce platelet aggregation. Collagenwas used as an agonist for assaying adenosine triphosphate release, thromboxane B2, serotonin,cyclic adenosine monophosphate, and cyclic guanosine monophosphate (cGMP) release. Results: When treated ex vivo, PDF not only inhibited ADP and collagen-induced platelet aggregation,but also upregulated cGMP levels and reduced platelet adhesion to fibronectin. Furthermore, it alsoinhibited Akt phosphorylation induced by collagen treatment. Panaxadiol fraction did not exert any antiplateletactivity in vitro, whereas PTF exhibited potent anti-platelet activity, inhibiting ADP, collagen, andthrombin-induced platelet aggregation, but significantly elevated levels of cGMP. Conclusion: Our study showed that in vitro and ex vivo PDF and PTF treatments exhibited differentpotency levels, indicating possible metabolic conversions of ginsenosides, which altered the content ofginsenosides capable of preventing platelet aggregation.

Ginkgolide C Inhibits Platelet Aggregation in cAMP- and cGMP-Dependent Manner by Activating MMP-9

Cho, Hyun-Jeong,Shon, Yun-Hee,Nam, Kyung-Soo Pharmaceutical Society of Japan 2007 Biological & pharmaceutical bulletin Vol.30 No.12

<P>In this report, we investigated the effect of ginkgolide C (GC) from <I>Ginkgo biloba</I> leaves in collagen (10 μg/ml)-stimulated platelet aggregation. It has been known that matrix metalloproteinase-9 (MMP-9) is released from human platelets, and that it significantly inhibited platelet aggregation stimulated by collagen. Zymographic analysis confirmed that pro-MMP-9 (92-kDa) was activated by GC to form an activated MMP-9 (86-kDa) on gelatinolytic activities. And then, GC dose-dependently inhibited platelet aggregation, intracellular Ca<SUP>2+</SUP> mobilization, and thromboxane A<SUB>2</SUB> (TXA<SUB>2</SUB>) formation in collagen-stimulated platelets. In addition, GC significantly increased the formation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), which have an anti-platelet function in both resting and collagen-stimulated platelets. Therefore, we demonstrate that the inhibitory effect of GC on platelet aggregation might be involved into the following pathways. GC may increase intracellular cAMP and cGMP production and MMP-9 activity, inhibit intracellular Ca<SUP>2+</SUP> mobilization and TXA<SUB>2</SUB> production, thereby leading to inhibition of platelet aggregation. These results strongly indicate that GC is a potent inhibitor of collagen-stimulated platelet aggregation. It may be a suitable tool for a negative regulator during platelet activation.</P>

Inhibitory Effect of Ginkgolide B on Platelet Aggregation in a cAMP- and cGMP-dependent Manner by Activated MMP-9

Cho, Hyun-Jeong,Nam, Kyung-Soo Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.5

Extracts from the leaves of the Ginkgo biloba are becoming increasingly popular as a treatment that is claimed to reduce atherosclerosis, coronary artery disease, and thrombosis. In this study, the effect of ginkgolide B (GB) from Ginkgo biloba leaves in collagen (10 ${\mu}g/ml$)-stimulated platelet aggregation was investigated. It has been known that human platelets release matrix metallo-proteinase-9 (MMP-9), and that it significantly inhibited platelet aggregation stimulated by collagen. Zymographic analysis confirmed that pro-MMP-9 (92-kDa) was activated by GB to form an MMP-9 (86-kDa) on gelatinolytic activities. And then, activated MMP-9 by GB dose-dependently inhibited platelet aggregation, intracellular $Ca^{2+}$ mobilization, and thromboxane $A_2$ ($TXA_2$) formation in collagen-stimulated platelets. Activated MMP-9 by GB directly affects down-regulations of cyclooxygenase-1 (COX-1) or $TXA_2$ synthase in a cell free system. In addition, activated MMP-9 significantly increased the formation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), which have the anti-platelet function in resting and collagen-stimulated platelets. Therefore, we suggest that activated MMP-9 by GB may increase the intracellular cAMP and cGMP production, inhibit the intracellular $Ca^{2+}$ mobilization and $TXA_2$ production, thereby leading to inhibition of platelet aggregation. These results strongly indicate that activated MMP-9 is a potent inhibitor of collagen-stimulated platelet aggregation. It may act a crucial role as a negative regulator during platelet activation.

전혈을 이용한 혈소판 응집능검사의 임상적 유용성에 관한 고찰

하영주,채석래,전회선,김현태 大韓血液學會 1993 大韓血液學會誌 Vol.28 No.1