혈관확장제의 응용 : 마취과영역에서

조영일 인제대학교 1981 仁濟醫學 Vol.2 No.2

Key words: Vasodilator. Stroke Volume. Preload. Afterload. Coronary Blood Flow.

Inhibitory effects of isoscopoletin on thrombus formation via regulation of cyclic nucleotides in collagen-induced platelets

이동하 한국응용생명화학회 2020 Journal of Applied Biological Chemistry (J. Appl. Vol.63 No.3

An essential component of the hemostatic process during vascular damage is platelet activation. However, many cardiovascular diseases, such as atherosclerosis, thrombosis, and myocardial infarction, can develop due to excessive platelet activation. Isoscopoletin, found primarily in plant roots of the genus Artemisia or Scopolia, has been studied to demonstrate potential pharmacological effects on Alzheimer's disease and anticancer, but its mechanisms and role in relation to thrombus formation and platelet aggregation have not yet been discovered. This research investigated the effect of isoscopoletin on collagen-induced human platelet activation. As a result, isoscopoletin strongly increased cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels in a concentration-dependent manner. In addition, isoscopoletin greatly phosphorylated inositol 1,4,5-triphosphate receptor (IP3R) and vasodilator-stimulated phosphoprotein (VASP), known substrates of cAMP-dependent kinase and cGMP dependent kinase. Phosphorylation of IP3R by isoscopoletin induced Ca2+ inhibition from the dense tubular system Ca2+ channels, and VASP phosphorylation was involved in fibrinogen binding inhibition by inactivating αIIb/β3 in the platelet membrane. Isoscopoletin finally reduced thrombin-induced fibrin clot production and finally reduced thrombus formation. Therefore, this research suggests that isoscopoletin has strong antiplatelet effects and is likely to be helpful for thrombotic diseases involving platelets by acting as a prophylactic and therapeutic agent.

Nitric Oxide Donor 첨가가 구리 결핍 배아의 발달과 Nitric Oxide 하위 신호전달체계에 미치는 영향

양수진(Yang Soo Jin),Keen Carl L,Uriu-Adams,Janet Y 韓國營養學會 2008 Journal of Nutrition and Health Vol.41 No.8

본 연구는 착상 후 단계의 쥐 배아와 난황낭을 대상으로 구리 결핍이 NO 하부 신호전달체계에 영향을 주는지를 알아보기 위한 것으로, 연구 결과는 다음과 같이 요약할 수 있다. 첫째, 구리 결핍은 정상적인 배아 및 난황낭 발달을 억제하고, NO의 생물학적 이용도와 아세틸콜린에 대한 NO dose-response를 낮추었다. 둘째, 구리 결핍은 NO의 하부 신호전달 물질인 cGMP 수준을 감소시켰으나, NO/cGMP하부 신호전달체계 표적 중 하나인 P-VASP에는 영향을 미치지 않았다. 셋째, 구리 결핍 배양액에 NO donor를 첨가하는 것은 구리 결핍 배아와 난황낭의 기형 발생 빈도를 구리 정상군과 비슷한 수준으로 개선시켰다. 넷째, NO donor 첨가는 구리 결핍군에서 감소되었던 cGMP의 농도를 유의적으로 증가시켰지만, P-VASP에는 영향을 미치지 않았다. 상기 연구 결과들은 구리 결핍으로 인한 NO의 생물학적 이용도의 감소가 기형발생의 주요 발생 기전이라는 것을 뒷받침하고 있다. 또한, 임상적으로 임신 기간 중 적절한 구리 섭취의 중요성을 강조한다. One suggested mechanism underlying copper (Cu) deficiency teratogenicity is a low availability of nitric oxide (NO), a signaling molecule which is essential in developmental processes. Increased superoxide anions secondary to decreased activities of Cu-zinc superoxide dismutase (Cu-Zn SOD) in Cu deficiency can interact with NO to form peroxynitrite, which can nitrate proteins at tyrosine residues. In addition, peroxynitrite formation can limit NO bioavailability. We previously reported low NO availability and increased protein nitration in Cu deficient (Cu-) embryos. In the current study, we tested whether Cu deficiency alters downstream signaling of NO by assessing cyclic GMP (cGMP) and phosphorylated vasodilator-stimulating phosphoprotein (VASP) levels, and whether NO supplementation can affect these targets as well as protein nitration. Gestation day 8.5 embryos from Cu adequate (Cu+) or Cu- dams were collected and cultured in either Cu+ or Cu- media for 48 hr. A subset of embryos was cultured in Cu- media supplemented with a NO donor (DETA/NONOate; 20 μM) and/or Cu-Zn SOD. Cu-/Cu- embryos showed a higher incidence of embryonic and yolk sac abnormalities, low NO availability, blunted dose-response in NO concentrations to increasing doses of acetylcholine, low mRNA expression of endothelial nitric oxide synthase (eNOS), increased levels of 3-nitrotyrosine (3-NT) compared to Cu+/Cu+ controls. cGMP concentrations tended to be low in Cu-/Cu- embryos, and they were significantly lower in Cu-/Cu- yolk sacs than in controls. Levels of phosphorylated VASP at serine 239 (P-VASP) were similar in all groups. NO donor supplementation to the Cu- media ameliorated embryonic and yolk sac abnormalities, and resulted in increased levels of cGMP without altering levels of P-VASP and 3-NT. Taken together, these data support the concept that Cu deficiency limits NO availability and alters NO/cGMP-dependent signaling in Cu- embryos and yolk sacs, which contributes to Cu deficiency-induced abnormal development.

Inhibitory effects of total saponin from Korean red ginseng via vasodilator-stimulated phosphoprotein-Ser<SUP>157</SUP> phosphorylation on thrombin-induced platelet aggregation

Dong-Ha Lee,Hyun-Jeong Cho,Hyun-Hong Kim,Man Hee Rhee,Jin-Hyeob Ryu,Hwa-Jin Park 고려인삼학회 2013 Journal of Ginseng Research Vol.37 No.2

In this study, we have investigated the effects of total saponin from Korean red ginseng (TSKRG) on thrombin-induced platelet aggregation. TSKRG dose-dependently inhibited thrombin-induced platelet aggregation with IC50 value of about 81.1 μg/mL. In addition, TSKRG dose-dependently decreased thrombin-elevated the level of cytosolic-free Ca<SUP>2+</SUP> ([Ca<SUP>2+</SUP>]i), one of aggregation-inducing molecules. Of two Ca<SUP>2+</SUP>-antagonistic cyclic nucleotides as aggregation-inhibiting molecules, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), TSKRG significantly dose-dependently elevated intracellular level of cAMP, but not cGMP. In addition, TSKRG dose-dependently inhibited thrombin-elevated adenosine triphosphate (ATP) release from platelets. These results suggest that the suppression of [Ca<SUP>2+</SUP>]i elevation, and of ATP release by TSKRG are associated with upregulation of cAMP. TSKRG elevated the phosphorylation of vasodilator-stimulated phosphoprotein (VASP)-Ser<SUP>157</SUP>, a cAMP-dependent protein kinase (A-kinase) substrate, but not the phosphorylation of VASP-Ser<SUP>239</SUP>, a cGMPdependent protein kinase substrate, in thrombin-activated platelets. We demonstrate that TSKRG involves in increase of cAMP level and subsequent elevation of VASP-Ser<SUP>157</SUP> phosphorylation through A-kinase activation to inhibit [Ca<SUP>2+</SUP>]i mobilization and ATP release in thrombin-induced platelet aggregation. These results strongly indicate that TSKRG is a beneficial herbal substance elevating cAMP level in thrombin-platelet interaction, which may result in preventing of platelet aggregation-mediated thrombotic diseases.

Inhibitory effects of total saponin from Korean red ginseng via vasodilator-stimulated phosphoprotein-Ser157 phosphorylation on thrombin-induced platelet aggregation

이동하,이만휘,조현정,박화진,김현홍 고려인삼학회 2013 Journal of Ginseng Research Vol.37 No.2

In this study, we have investigated the effects of total saponin from Korean red ginseng (TSKRG) on thrombin-induced platelet aggregation. TSKRG dose-dependently inhibited thrombin-induced platelet aggregation with IC50 value of about 81.1μg/mL. In addition, TSKRG dose-dependently decreased thrombin-elevated the level of cytosolic-free Ca2+ ([Ca2+]i), one of aggregation-inducing molecules. Of two Ca2+-antagonistic cyclic nucleotides as aggregation-inhibiting molecules, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), TSKRG significantly dose-dependently elevated intracellular level of cAMP, but not cGMP. In addition, TSKRG dose-dependently inhibited thrombin-elevated adenosine triphosphate (ATP) release from platelets. These results suggest that the suppression of [Ca2+]i elevation, and of ATP release by TSKRG are associated with upregulation of cAMP. TSKRG elevated the phosphorylation of vasodilator-stimulated phosphoprotein (VASP)-Ser157, a cAMP-dependent protein kinase (A-kinase) substrate, but not the phosphorylation of VASP-Ser239, a cGMPdependent protein kinase substrate, in thrombin-activated platelets. We demonstrate that TSKRG involves in increase of cAMP level and subsequent elevation of VASP-Ser157 phosphorylation through A-kinase activation to inhibit [Ca2+]i mobilization and ATP release in thrombin-induced platelet aggregation. These results strongly indicate that TSKRG is a beneficial herbal substance elevating cAMP level in thrombin-platelet interaction, which may result in preventing of platelet aggregation-mediated thrombotic diseases.

콜라겐 유도의 혈소판에서 사이클릭 뉴클레오티드의 조절을 통한 Scoparone의 혈전 형성 억제 효과

이동하 한국식품영양과학회 2020 한국식품영양과학회지 Vol.49 No.1

혈소판 활성화는 혈관 손상에 대한 지혈 과정에 필수적이다. 그러나 과도한 혈소판 활성화는 죽상동맥경화증, 혈전증 및 심근경색을 포함한 여러 심혈관 질환을 유발할 수 있다. Scoparone은 Artemisia 또는 Scopolia 속의 뿌리에서 흔히 발견되며 면역억제 및 혈관 이완을 포함한 약리학적인 특성에 대해 연구되었지만, 항혈소판 효과에 대해서는 아직 보고된 바가 없다. 본 연구는 scoparone이 collagen에 의해 유도한 사람 혈소판 활성화에 미치는 영향을 연구하였다. 그 결과로, scoparone은 농도 의존적으로 cyclic adenosine monophosphate(cAMP)와 cyclic guanosine monophosphate(cGMP) 수준을 유의하게 증가시켰다. 또한 scoparone은 cAMP 의존성 kinase(PKA) 및 cGMP 의존성 kinase(PKG)의 기질로 작용하는 inositol 1,4,5-triphosphate receptor(IP3R) 및 vasodilator-stimulated phosphoprotein(VASP)을 유의하게 인산화시켰다. Scoparone에 의한 IP3R의 인산화는 dense tubular system의 Ca2+ 채널로부터 동원되는 Ca2+ 억제를 유발하였고, VASP의 인산화는 혈소판 막에 있는 αIIb/β3의 불활성화를 일으켜 fibrinogen 결합을 저해함으로써 혈소판 활성을 억제하는 데 관여하였다. Scoparone은 thrombin-유도의 fibrin clot 생성을 억제함으로써 최종적으로 혈전 형성을 감소시켰다. 그러므로 본 연구에서는 scoparone이 강력한 항혈소판 효과를 가지며 혈소판이 유래하는 혈전성 질환의 예방 및 치료제로서 가능성이 있음을 제안한다. Platelet activation is essential for the hemostatic process for vascular damage. Excessive platelet activation, however, can lead to several cardiovascular diseases, including atherosclerosis, thrombosis, and myocardial infarction. Scoparone is found in the roots of the genus Artemisia or Scopolia. The pharmacological properties of this compound, including immunosuppression and vascular relaxation, have been studied, but no antiplatelet effects have been reported. This study examined the activities of scoparone on collagen-induced human platelet activation. Scoparone increased the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in a concentration-dependent manner. In addition, scoparone phosphorylated the inositol 1,4,5-triphosphate receptor (IP3R) and vasodilator-stimulated phosphoprotein (VASP) that act as substrates for cAMP-dependent kinase (PKA) and cGMP-dependent kinase (PKG). The phosphorylation of IP3R by scoparone-inhibited the Ca2+ mobilized from the Ca2+ channel of the dense tubular system. Moreover, the phosphorylation of VASP was involved in the inhibition of fibrinogen binding by inactivating αIIb/β3 in the platelet membrane. Scoparone finally reduced the level of thrombin-induced fibrin clot production and reduced thrombus formation. Therefore, scoparone has strong antiplatelet activity and is a potential prophylactic and therapeutic agent for thrombotic diseases from which platelets are derived.

Inhibitory effects of total saponin from Korean red ginseng via vasodilator-stimulated phosphoprotein-Ser¹⁵⁷ phosphorylation on thrombin-induced platelet aggregation

Lee, Dong-Ha,Cho, Hyun-Jeong,Kim, Hyun-Hong,Rhee, Man Hee,Ryu, Jin-Hyeob,Park, Hwa-Jin The Korean Society of Ginseng 2013 Journal of Ginseng Research Vol.37 No.2

In this study, we have investigated the effects of total saponin from Korean red ginseng (TSKRG) on thrombin-induced platelet aggregation. TSKRG dose-dependently inhibited thrombin-induced platelet aggregation with $IC_{50}$ value of about 81.1 ${\mu}g/mL$. In addition, TSKRG dose-dependently decreased thrombin-elevated the level of cytosolic-free $Ca^{2+}$ ($[Ca^{2+}]_i$), one of aggregation-inducing molecules. Of two $Ca^{2+}$-antagonistic cyclic nucleotides as aggregation-inhibiting molecules, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), TSKRG significantly dose-dependently elevated intracellular level of cAMP, but not cGMP. In addition, TSKRG dose-dependently inhibited thrombin-elevated adenosine triphosphate (ATP) release from platelets. These results suggest that the suppression of $[Ca^{2+}]_i$ elevation, and of ATP release by TSKRG are associated with upregulation of cAMP. TSKRG elevated the phosphorylation of vasodilator-stimulated phosphoprotein (VASP)-$Ser^{157}$, a cAMP-dependent protein kinase (A-kinase) substrate, but not the phosphorylation of VASP-$Ser^{239}$, a cGMP-dependent protein kinase substrate, in thrombin-activated platelets. We demonstrate that TSKRG involves in increase of cAMP level and subsequent elevation of VASP-$Ser^{157}$ phosphorylation through A-kinase activation to inhibit $[Ca^{2+}]_i$ mobilization and ATP release in thrombin-induced platelet aggregation. These results strongly indicate that TSKRG is a beneficial herbal substance elevating cAMP level in thrombin-platelet interaction, which may result in preventing of platelet aggregation-mediated thrombotic diseases.

U46619 유도의 혈소판에서 Cyclic Nucleotides 조절을 통한 Isoscopoletin의 혈전생성 억제효과

이동하 한국생약학회 2021 생약학회지 Vol.52 No.1

During blood vessel damage, an essential step in the hemostatic process is platelet activation. However, it is important to properly control platelet activation, as various cardiovascular diseases, such as stroke, atherosclerosis, and myocardial infarction, are also caused by excessive platelet activation. Found primarily in the roots of plants of the genus Artemisia or Scopolia, isoscopoletin has been studied to demonstrate its potential pharmacological effects against Alzheimer's disease and anticancer, but the mechanisms and roles involved in thrombus formation and platelet aggregation are insufficient. This study investigated the effect of isoscopoletin on U46619-induced human platelet activation. As a result, isoscopoletin significantly increased the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) dose-dependently. In addition, isoscopoletin significantly phosphorylated inositol 1, 4, 5-triphosphate receptor (IP3R) and vasodilator-stimulated phosphprotein (VASP), which are known substrates for cAMP-dependent kinases and cGMP-dependent kinases. Phosphorylated IP3R by isoscopoletin inhibited Ca2+ mobilization from the dense tubular system Ca2+ channels to cytosol, and phosphorylated VASP was involved in the inhibition of fibrinogen binding through aIIb/β3 inactivation in the platelet membrane. Isoscopoletin finally reduced thrombin-induced fibrin clotting production. Therefore, this study suggests that isoscopoletin has a potent antiplatelet effect and may be helpful for platelet-related thrombotic diseases.

Cudrania Tricuspidata root extract (CTE) has an anti-platelet effect via cGMP-dependent VASP phosphorylation in human platelets

Ju-Ye Ro(노주예),Hyun-Jeong Cho(조현정) 한국산학기술학회 2019 한국산학기술학회논문지 Vol.20 No.12

꾸지뽕나무 뿌리 추출물은 항염, 항암, 항산화와 같은 효과를 갖는 많은 생리활성 물질을 포함하고 있다고 알려져 있다. 그러나 꾸지뽕나무 뿌리 추출물(이하 CTE)의 사람 혈소판 응집 억제 기전에 관하여는 알려진 바 없다. 본 연구에서는 CTE가 혈소판에 미치는 영향을 확인하고자 하였다. CTE는 collagen으로 유도한 혈소판 응집 반응에서 cyclooxygenase-1 활성을 억제하고, 세포 내 칼슘 농도를 낮추는 방식으로 thromboxane A2 생성을 억제하였다. 또한, phospholipase Cγ2와 syk의 인산화 반응을 억제하였으며, guanosine monophosphate (cGMP)에 의존적인 방식으로 vasodilator-stimulated phosphoprotein(VASP)의 Ser<SUP>239</SUP> 위치를 인산화하여 항혈소판 효과를 나타내었다. 또한, 고지방 식이로 혈소판 활성화를 유도한 랫드에서 CTE를 경구 투여 하였을 때, 간독성 없이 콜라겐으로 유도한 혈소판 응집반응을 thromboxane A2 생성을 억제함으로써 항혈소판 효과를 보였다. 이는 in vivo 와 in vitro 에서 같은 결과를 제시하고 있다. 결론적으로, CTE는 항혈소판 작용 및 심혈관계 질환 예방을 위한 천연물 유래의 안전하고, 새로운 물질임을 제시하는 바이다. Cudrania tricuspidata has been reported to have many biological activities, including anti-inflammatory, anti-cancer, and antioxidant properties. However, the effects of C. tricuspidata root extract (CTE) on human platelet aggregation induced by collagen as well as the signaling pathways involved remain unknown. In the present study, we investigated the effect of CTE on human platelets. CTE inhibited platelet aggregation via down-regulation of thromboxane A2 (TXA2) by blocking cyclooxygenase-1 (COX-1) activity and intracellular Ca<SUP>2+</SUP> mobilization in collagen-induced platelets. CTE also reduced the phosphorylation of phospholipase C (PLC) γ2 and syk. CTE regulated platelet aggregation via cyclic guanosine monophosphate (cGMP)-dependent phosphorylation of vasodilator-stimulated phosphoprotein (VASP) Ser239. In addition, administration of CTE (50 and 100 mg/kg) significantly reduced hyper-aggregated platelet aggregation by collagen (5 μg/mL) without hepatotoxicity in HFD (high fat diet)-fed rats. Taken together, these results suggest that CTE has anti-platelet effects both in vitro and in vivo. Therefore, CTE may be an effective therapeutic and preventive agent for cardiovascular disease, and is a safe and natural product.

Cudrania Tricuspidata root extract (CTE) has an anti-platelet effect via cGMP-dependent VASP phosphorylation in human platelets

노주예,조현정 한국산학기술학회 2019 한국산학기술학회논문지 Vol.20 No.12

Cudrania tricuspidata has been reported to have many biological activities, including anti-inflammatory, anti-cancer, and antioxidant properties. However, the effects of C. tricuspidata root extract (CTE) on human platelet aggregation induced by collagen as well as the signaling pathways involved remain unknown. In the present study, we investigated the effect of CTE on human platelets. CTE inhibited platelet aggregation via down-regulation of thromboxane A2 (TXA2) by blocking cyclooxygenase-1 (COX-1) activity and intracellular Ca2+ mobilization in collagen-induced platelets. CTE also reduced the phosphorylation of phospholipase C (PLC) γ2 and syk. CTE regulated platelet aggregation via cyclic guanosine monophosphate (cGMP)-dependent phosphorylation of vasodilator-stimulated phosphoprotein (VASP) Ser239. In addition, administration of CTE (50 and 100 mg/kg) significantly reduced hyper-aggregated platelet aggregation by collagen (5 μg/mL) without hepatotoxicity in HFD (high fat diet)-fed rats. Taken together, these results suggest that CTE has anti-platelet effects both in vitro and in vivo. Therefore, CTE may be an effective therapeutic and preventive agent for cardiovascular disease, and is a safe and natural product.