혈소판의 냉동보존 방법의 차이에 따른 활성도의 변화

박경화,김병수,배숙영,윤수영,설혜령,최종권,성화정,윤소영,최인근,오상철,서재홍,최철원,신상원,김열홍,김준석 大韓血液學會 2003 大韓血液學會誌 Vol.38 No.3

Characterization of the distinct mechanism of agonist-induced canine platelet activation

Chaudhary, Preeti K.,Kim, Soochong The Korean Society of Veterinary Science 2019 Journal of Veterinary Science Vol.20 No.1

<P>Platelet activation has a major role in hemostasis and thrombosis. Various agonists including adenosine diphosphate (ADP) and thrombin interact with G protein-coupled receptors (GPCRs) which transduce signals through various G proteins. Recent studies have elucidated the role of GPCRs and their corresponding G proteins in the regulation of events involved in platelet activation. However, agonist-induced platelet activation in companion animals has not been elucidated. This study was designed to characterize the platelet response to various agonists in dog platelets. We found that 2-methylthio-ADP-induced dog platelet aggregation was blocked in the presence of either P2Y<SUB>1</SUB> receptor antagonist MRS2179 or P2Y<SUB>12</SUB> receptor antagonist AR-C69931MX, suggesting that co-activation of both the P2Y<SUB>1</SUB> and P2Y<SUB>12</SUB> receptors is required for ADP-induced platelet aggregation. Thrombin-induced dog platelet aggregation was inhibited in the presence of either AR-C69931MX or the PKC inhibitor GF109203X, suggesting that thrombin requires secreted ADP to induce platelet aggregation in dog platelets. In addition, thrombin-mediated Akt phosphorylation was inhibited in the presence of GF109203X or AR-C69931MX, indicating that thrombin causes Gi stimulation through the P2Y<SUB>12</SUB> receptor by secreted ADP in dog platelets. Unlike human and murine platelets, protease-activated receptor 4 (PAR4)-activating peptide AYPGKF failed to cause dog platelet aggregation. Moreover, PAR1-activating peptide SFLLRN or co-stimulation of SFLLRN and AYPGKF failed to induce dog platelet aggregation. We conclude that ADP induces platelet aggregation through the P2Y<SUB>1</SUB> and P2Y<SUB>12</SUB> receptors in dogs. Unlike human and murine platelets, selective activation of the PAR4 receptor may be insufficient to cause platelet aggregation in dog platelets.</P>

혈소판제제의 저장기간에 따른 cytokine 변화와 수혈자 호중구 respiratory burst에 미치는 영향

임영애,박애자 중앙대학교 의과대학 의과학연구소 1997 中央醫大誌 Vol.22 No.4

Interleukin(IL)-8 generated from the stored platelets are known to be a cytokine to activate the neutrophils by priming effects, especially in patients with the primed neutrophils and to have a synergistic effect with other cytokines such as IL-6, IL-1β and tumor necrosis factor(TNF)-α. This study was conducted to evaluate the cytokine release from the stored platelets and their effects on neutrophil activation. Ten units of fresh platelet concentrates and ten units of platelet-pheresis products were obtained from healthy donors and kept on agitator at room temperature. Samples were collected from the above preparations on day 0, 1, 3, 5 and 7 of storage for counts of leukocyte and platelet and assays of IL-8, IL-6, IL-1β and TNF-α using enzyme immunoassay. Platelet poor plasmas obtained from on day 0, 1, 3, 5 and 7 of storage or recombinant human(고) IL-8 were incubated with neutrophils from healthy donors and patients to measure the neutrophil respiratory burst using 2',7'-dichlorofluoroscein diacetate by flowcytometry. The results were expressed as by Mean Channel Fluorescence(MCF) Ratio (MCF value of stored platelets/MCF value of negative control) and were summarized as below: 1. Mean leukocyte count of random platelet concentrates(1.23±0.32x10^9/L) was significantly higher than that of platelet-pheresis products(0.45±0.28 x10^9/L, P<0.05). 2. The concentrations of IL-8 and IL-6 were significantly higher in random platelet concentrates than in platelet-pheresis products. Especially, the concentration of IL-8 was progressively increased after 3 days of storage, depending on the storage time. 3. All of the MCF ratio of stored platelet preparations on day 1, 3, 5 and 7 with patient's or healthy donor's neutrophils were not significantly different from those of day 0(P>0.05). However mean MCF ratio in the patient group(1.125±0.363) was signifivantly higher than that of healthy donor group(1.024±0.274, P<0.05). 4. Priming effects of whole blood neutrophils were found with rhIL-8 when stimulated 10 ng/ml for 5 minutes incubation, however, 1 ng/ml, 5 ng/ml, 10 ng/ml, 100 ng/ml for 30 miniutes incubation(P<0.05). In conclusion, stored random platelet concentrates contained high leukocyte count may relate to non-hemolytic transfusion reactions by increment of concentrations of cytokines, which have synergistic effects to neutrophil activation in primed recipients. Therefore, especially massive transfusion of stored random platelet concentrates should be avoided in already primed patients to prevent nonhemolytic transfusion reactions due to activated neutrophils.

The Fibrinogen to Mean Platelet Volume Ratio Can Predict Overall Survival of Patients with Non-Metastatic Gastric Cancer

Song, Shubin,Cong, Xiliang,Li, Fengke,Xue, Yingwei The Korean Gastric Cancer Association 2018 Journal of gastric cancer Vol.18 No.4

Purpose: Fibrinogen and platelets have been reported to play important roles in tumorigenesis and cancer progression. The aim of this research was to investigate the combination of functions of fibrinogen, platelets, and mean platelet volume (MPV) in predicting the survival of patients with gastric cancer (GC). Materials and Methods: A retrospective study was conducted with 1,946 patients with GC and 299 patients with benign gastric tumor to analyze their fibrinogen, platelet, and MPV levels, and other clinicopathological characteristics along with their prognoses. Several indicators were evaluated along with fibrinogen, platelets, and MPV and their prognostic abilities were assessed. Univariate and multivariate survival analyses were conducted to determine the independent risk factors for overall survival. Results: Increased levels of fibrinogen, platelets, and MPV were observed with the progress of the GC stages. Elevated fibrinogen, platelets, and the combined indicators, including fibrinogen*MPV (FM), platelet*fibrinogen*MPV (PFM), fibrinogen/MPV (FMR), platelet*fibrinogen (PF), platelet*fibrinogen/MPV (PFMR), platelet*MPV (PM), and platelet/MPV (PMR), foreboded poor prognosis. Meanwhile fibrinogen and FMR can be considered as independent risk factors for overall survival in patients with non-metastatic GC. But these indicators can hardly predict survival of patients in stage IV. Conclusions: Elevated fibrinogen, platelets, and MPV levels were in accordance with advanced stages, and fibrinogen, platelet, and MPV, in combination, can be used to predict survival of patients with non-metastatic GC. FMR was an independent prognostic factor for overall survival of patients with GC.

Menadione의 대사체인 Menadione-Glutathione Conjugate(MEN-SG)가 흰쥐 혈소판에 미치는 세포독성의 평가 및 MEN-SG의 안정성에 관한 연구

서동철,정선화,이주영,김미정,정진호,Seo, Dong-Chul,Chung, Sun-Hwa,Lee, Joo-Young,Kim, Mee-Jeong,Chung, Jin-Ho 한국독성학회 1995 Toxicological Research Vol.11 No.2

Menadione-ghitathione conjugate (MEN-SG), a metabolite of menadione, is known to be a redoxcycler in rat hepatocyte subcellular fraction. Therefore, it was assumed that MEN-SG could exert cytotoxlclty to ral platelets, another target tissue of menadione. We first synthesized MEN-SG, the identity of which was verified by mass, $^1{H}$-NMR and UV-visible spectra. In addition, the stability of MEN-SG was investigated in biological assay system. MEN-SG was degraded in a time-dependent manner in DMSO which had been used as a vehicle and thus, tris-HCl buffer was used as a vehicle of MEN-SG despite the low solubility in it. Perchloric acid as well as platelets itself did not affect the stability of MEN-SG. Our next attempt was the evaluation of cytotoxicity of MEN-SG in rat platelets. MEN-SG did not induce cytotoxicity to rat platelets measured by two different methods, LDH release and turbidity changes. The extents of oxygen consumption by MEN-SG in intact platelets were significantly lower than those by menadione, though it had been observed that oxygen consumptions by menadione and MENSG were similar in subcellular fractioas of platelets. These results suggest that MEN-SG is not toxic to rat platelets despite its redox cycling capacity and glutathione conjugation reaction of menadione could be regarded as a detoxification process.

Chemical-Induced Cytotoxicity in Platelet Rich Plasma Isolated from Rats

Seung, Sang-Ae,Chung, Seung-Min,Lee, Sun-Koo,Lee, Joo-Young,Kim, Jeong-Sun,Chung, Jin-Ho Korean Society of ToxicologyKorea Environmental Mu 1997 Toxicological Research Vol.13 No.3

The elevation of intracellular calcium in various tissues due to oxidative stress induced by either menadione or adriamycin has been well documented. The increase of calcium level in platelets results in aggregation of platelets. To test the hypothesis that chemically induced calcium elevations can play a role in platelet aggregation, we have studied the effects of menadione and adriamycin on aggregation of platelets isolated from female rats. Treatment with menadione and adriamycin to platelet rich plasma (PRP) appeared to induce platelet aggregations up to 60%, as determined by aggregometry. However, exposure of PRP to rnenadione or adriamycin led to a loss of viability, as measured by lactate dehydrogenase (LDH) leakage. Morphological studies of platelets revealed that, when PRP was treated with menadione, aggregates of platelets were not observed and the numbers of platelets were decreased significantly. This suggests that menadione and adriamycin decreased turbidity by inducing platelet lysis rather than platelet aggregation. These cellular toxicities induced by menadione or adriamycin was not correlated with oxygen consumption rate but with depletion of protein thiols, suggesting that protein thiols might play an important role in chemical-induced platelet toxicity.

The Fibrinogen to Mean Platelet Volume Ratio Can Predict Overall Survival of Patients with Non-Metastatic Gastric Cancer

Shubin Song,Xiliang Cong,Fengke Li,Yingwei Xue 대한위암학회 2018 Journal of gastric cancer Vol.18 No.4

Purpose: Fibrinogen and platelets have been reported to play important roles in tumorigenesis and cancer progression. The aim of this research was to investigate the combination of functions of fibrinogen, platelets, and mean platelet volume (MPV) in predicting the survival of patients with gastric cancer (GC). Materials and Methods: A retrospective study was conducted with 1,946 patients with GC and 299 patients with benign gastric tumor to analyze their fibrinogen, platelet, and MPV levels, and other clinicopathological characteristics along with their prognoses. Several indicators were evaluated along with fibrinogen, platelets, and MPV and their prognostic abilities were assessed. Univariate and multivariate survival analyses were conducted to determine the independent risk factors for overall survival. Results: Increased levels of fibrinogen, platelets, and MPV were observed with the progress of the GC stages. Elevated fibrinogen, platelets, and the combined indicators, including fibrinogen*MPV (FM), platelet*fibrinogen*MPV (PFM), fibrinogen/MPV (FMR), platelet*fibrinogen (PF), platelet*fibrinogen/MPV (PFMR), platelet*MPV (PM), and platelet/ MPV (PMR), foreboded poor prognosis. Meanwhile fibrinogen and FMR can be considered as independent risk factors for overall survival in patients with non-metastatic GC. But these indicators can hardly predict survival of patients in stage IV. Conclusions: Elevated fibrinogen, platelets, and MPV levels were in accordance with advanced stages, and fibrinogen, platelet, and MPV, in combination, can be used to predict survival of patients with non-metastatic GC. FMR was an independent prognostic factor for overall survival of patients with GC.

혈소판의 냉동보존에 미치는 일산화질소의 효과

이재현,김정태,조용곤 대한진단검사의학회 2008 Annals of Laboratory Medicine Vol.28 No.2

Background : To determine whether nitric oxide (NO) could inhibit activation of platelets stored in a cold or frozen state, we measured platelet P-selectin expression and platelet-bound fibrinogen in platelet-rich plasma (PRP) with S-nitrosoglutathione (GSNO) (Sigma, USA) by flow cytometry. Methods : PRP was prepared by centrifuging venous blood collected in a 3.2% sodium citrate tube from 10 healthy donors. It was aliquotted into 4 groups (no cryoprotectant, GSNO, GSNO/ dimethyl sulfoxide [DMSO] [Sigma], and DMSO), and stored at room, cold and freezing temperatures for 24 hrs. We performed a flow cytometric analysis of all specimens stained with FITC-fibrinogen and PE-CD62P monoclonal antibodies (Becton Dickinson, USA). The results were compared according to the storage temperature and agonist among 4 groups. Results : GSNO inhibited significantly the activation of frozen platelets, but not in the presence of DMSO. GSNO was also shown to preserve the aggregability of frozen platelets because in the presence of GSNO the delta percent change of P-selectin expression and fibrinogen binding of frozen platelets increased significantly irrelevant to DMSO. Conclusions : GSNO inhibited the activation of frozen platelets and preserved the platelet aggregability; therefore, it may be used as a protectant for platelet cryopreservation. (Korean J Lab Med 2008;28:136-43)

Thapsigargin Induces Platelet Aggregation, thereby Releases Lactate Dehydrogenase from Rat Platelets

Ji Sue Baik,You Na Seo,Man Hee Rhee,Moon-Taek Park,Sung Dae Kim 대한의생명과학회 2021 Biomedical Science Letters Vol.27 No.3

Thapsigargin (TG), a sarco/endoplasmic reticulum (ER) Ca<SUP>2+</SUP>-ATPase (SERCA) inhibitor, has been widely used as an agonist for platelet aggregation for decades. In this study, we investigated the effect of TG on the release of lactate dehydrogenase (LDH) for platelets and elucidated its mechanism. Platelet LDH release and platelet aggregation were increased by TG treatment; 1,000 nM of TG induced the complete lysis of platelets. Other agonists such as collagen (2.5 μg/mL), thrombin (0.1 U/mL), and ADP (10 mM) did not induce significant platelet LDH release despite platelet aggregation. Finally, we investigated the effects of pharmacological inhibitors on TG-induced platelet aggregation and LDH release. SP600125, a JNK inhibitor, and LY294002, a PI-3K inhibitor, inhibited TG-induced platelet LDH release but not platelet aggregation. Forskolin, an adenylyl cyclase activator, also inhibited LDH release without affecting platelet aggregation by TG. These results suggest that the TG-induced platelet aggregation was accompanied by LDH release but regulated by a different signaling pathway.

메나디온에 의한 혈소판 내 칼슘 변화측정시 형광 색소 사용의 문제점

정선화(Sun Hwa Chung),이무열(Moo Yeol Lee),이주영(Joo Young Lee),정승민(Seung Min Chung),정진호(Jin Ho Chung) 대한약학회 1997 약학회지 Vol.41 No.6

It has been reported that dose-dependent Ca2+ increase by menadione in platelets could be measured by fluorescent dye, quin-2. The problems will be described here relating to measuring Ca2+ in menadione-exposed platelets using fura-2 and fluo-3, widely used fluorescent indicators. Additions of menadione to fura-2 loaded platelets and their lysates resulted in marked reduction in fluorescence intensity at both 340nm (Ca2+-unbound form) excitation wavelengths. Fura-2 excitation spectra were overlapped with UV-visible absorption spectra of menadione, suggesting that light absorption by menadione itself could quench fluorescence generated by fura-2. Next approach was to use fluo-3 which has the higher wavelength (490nm) of excitation. Previous work demonstrated that treatment with probenecid to platelets was required to prevent fluo-3 dye leakage. However, probenecid itself was proven to be inadequate to measure the concentration of intracellular Ca2+ by reducing menadione-induced cytotoxicity in platelets. Our results suggest that it is not feasible to measure Ca2+ in platelets by using fura-2 and fluo-3 in the presence of probenecid, and cautions should be taken to measure changes of intracellular Ca2+ levels by fluorescent dyes following chemical exposure.