Role of glycine-containing oligopeptides in blood cells aggregation stimulated by adrenaline

2017 ◽  
Author(s):  
М. Голубева
Keyword(s):  
Body Weight ◽  
Platelet Aggregation ◽  
Blood Cells ◽  
Regulatory Peptides ◽  
Pituitary Hormones ◽  
New Drugs ◽  
Male Rats ◽  
Current Interest

Введение. Изменение реологических свойств крови характерно для различных заболеваний, многие из которых связаны с нарушением реологии и, прежде всего, с изменением агрегации эритроцитов. Целью исследования было сравнение влияния малых регуляторных пептидов, являющихся фрагментами нейрогормонов, на агрегацию эритроцитов и тромбоцитов под действием адреналина в экспериментах in vitro. Материалы и методы. Эксперименты проводили на белых беспородных крысах-самцах, массой тела 180-200 г. Использовали пептиды, представляющие собой С-концевые фрагменты вазопрессина (Pro-Arg-Gly-NH2) и окситоцина (Pro-Leu-Gly-NH2). Результаты. Показано, что малые регуляторные пептиды, являющиеся продуктами протеолиза нейропептидов, оказывают существенное влияние на агрегатное состояние клеток крови. При сравнении влияния пептидов на агрегацию клеток крови, стимулированную адреналином, установлено, что фрагмент вазопрессина Pro-Arg-Gly-NH2 вызывал достоверное усиление агрегации как эритроцитов, так и тромбоцитов; тогда как фрагмент окситоцина Pro-Leu-Gly-NH2 ингибировал только агрегацию эритроцитов, не изменяя агрегации тромбоцитов. Заключение. Изучение путей поэтапного протеолиза пептидов может привести к разработке новых препаратов для направленной коррекции различных нарушений в организме, поэтому изучение эффектов С-концевых фрагментов гипофизарных гормонов на гемостаз является актуальным. Introduction. The changing of blood rheological properties is typical for various diseases; many of them are associated with rheology disorder and primarily with change of erythrocytes aggregation. The aim was to compare the effect of small regulatory peptides (fragments of neurohormones) on the aggregation of erythrocytes and platelets under adrenaline action in experiments in vitro. Materials and methods. Experiments were conducted on white outbred male rats, body weight 180-200 g. We used 2 peptides – C-terminal fragments of vasopressin (Pro-Arg-Gly-NH2) and oxytocin (Pro-Leu-Gly-NH2). Results. It was shown that small regulatory peptides (they are products of neuropeptides proteolysis) had a significant effect on blood cells aggregation. We compared the peptides effect on blood cells aggregation stimulated by adrenaline. It was found that vasopressin fragment Pro-Arg-Gly-NH2 significantly increased both erythrocytes and platelets aggregation, while oxytocin fragment Pro-Leu-Gly-NH2 inhibited only erythrocytes aggregation without changing of platelet aggregation. Conclusion. Investigation of phased peptides proteolysis may result in the development of new drugs for targeted correction of various disturbances. So it is of current interest to study the effects of C-terminal fragments of pituitary hormones on hemostasis.

Peptide correction of prothrombotic erythrocytes effects on platelet aggregation

2019 ◽  
Author(s):  
М.Г. Голубева
Keyword(s):  
Platelet Aggregation ◽  
Blood Cells ◽  
Thrombus Formation ◽  
Regulatory Peptides ◽  
Rat Plasma ◽  
New Drugs ◽  
In Vitro Experiments ◽  
Terminal Fragment ◽  
Thrombotic Complications

Введение. В патогенезе многих заболеваний важную роль играют изменения функции гемостаза и нарушение реологических свойств крови. Поиск новых лекарственных препаратов, влияющих на взаимодействие форменных элементов крови в процессе тромбообразования, является одной из актуальных задач современной гематологии. Цель исследования: сравнение влияния малых регуляторных пептидов, являющихся фрагментами нейрогормонов, на взаимодействие эритроцитов с тромбоцитами при их агрегации под действием адреналина в экспериментах in vitro. Материалы и методы. Исследовали пептиды, представляющие собой С-концевые фрагменты нейрогормонов: Pro-Arg-Gly-NH2 — фрагмент вазопрессина, Pro-Leu-Gly-NH2 — фрагмент окситоцина. В опытах in vitro пептиды в концентрации 10–4–10–6 М добавляли к пулу богатой тромбоцитами плазмы (БТП) крыс или к смеси БТП с эритроцитарной массой, разведенной 1:1000 физиологическим раствором, или к отмытым эритроцитам и тромбоцитам и их смеси, и определяли изменение агрегации под действием адреналина в конечной концентрации 0,02 ммоль/л. Агрегацию эритроцитов и тромбоцитов регистрировали на агрегометре. Результаты. Установлено, что С-концевой фрагмент вазопрессина обладает более ярко выраженным антиагрегантным действием, чем фрагмент окситоцина, причем это было установлено как в БТП, так и при использовании отмытых клеток крови. Ингибирующее действие фрагмента Pro-Arg-Gly-NH2 сохранялось и на фоне предварительного усиления агрегации тромбоцитов эритроцитами. С-концевой фрагмент окситоцина практически не влиял на агрегацию тромбоцитов. Заключение. Использование малых регуляторных пептидов позволяет снижать агрегационный эффект эритроцитов, улучшая тем самым терапию при тромботических осложнениях. Introduction. Hemostasis changes and disturbances of blood rheological properties play the important role in pathogenesis of many diseases. One of the actual problems of modern hematology is new drugs investigation for aff ecting on blood cells interaction during thrombus formation. Aim: to compare the eff ect of small regulatory peptides (fragments of neurohormones) on the interaction of erythrocytes with platelets under their aggregation by adrenaline in in vitro experiments. Materials and methods. We studied 2 peptides (C-terminal fragments of neurohormones): Pro-Arg-Gly-NH2 — fragment of vasopressin, Pro-Leu-Gly-NH2 — fragment of oxytocin. In in vitro experiments we added peptides (in concentration of 10–4–10–6 М) to a pool of platelet-rich rat plasma (PRP) or to a mixture of PRP with erythrocyte mass diluted 1:1000 by saline solution, or to washed erythrocytes and platelets and their mixture, and determined the aggregation changes under adrenalin in a final concentration of 0.02 mmol/L. Erythrocytes and platelets aggregation was recorded by aggregometer. Results. We found that C-terminal fragment of vasopressin has a more pronounced antiplatelet eff ect than the oxytocin fragment, and it was found both in PRP and with the use of washed blood cells. The inhibitory eff ect of Pro-Arg-Gly-NH2 fragment also remained under preliminary increasing of platelet aggregation by erythrocytes. C-terminal fragment of oxytocin practically had no effect on platelet aggregation. Conclusion. Use of small regulatory peptides helps to reduce the aggregation eff ect of erythrocytes, thereby improving therapy of thrombotic complications.

scholarly journals Prolactin and Male Fertility: The Long and Short Feedback Regulation

2009 ◽  
Vol 2009 ◽  
pp. 1-13 ◽  
Author(s):  
M. K. Gill-Sharma
Keyword(s):  
Chromatin Condensation ◽  
Feedback Regulation ◽  
Pituitary Hormones ◽  
Feedback Mechanism ◽  
Reproductive Hormones ◽  
Male Rats ◽  
Normal Men

In the last 20 years, a pituitary-hypothalamus tissue culture system with intact neural and portal connections has been developed in our lab and used to understand the feedback mechanisms that regulate the secretions of adenohypophyseal hormones and fertility of male rats. In the last decade, several in vivo rat models have also been developed in our lab with a view to substantiate the in vitro findings, in order to delineate the role of pituitary hormones in the regulation of fertility of male rats. These studies have relied on both surgical and pharmacological interventions to modulate the secretions of gonadotropins and testosterone. The interrelationship between the circadian release of reproductive hormones has also been ascertained in normal men. Our studies suggest that testosterone regulates the secretion of prolactin through a long feedback mechanism, which appears to have been conserved from rats to humans. These studies have filled in a major lacuna pertaining to the role of prolactin in male reproductive physiology by demonstrating the interdependence between testosterone and prolactin. Systemic levels of prolactin play a deterministic role in the mechanism of chromatin condensation during spermiogenesis.

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes
Keyword(s):  
Platelet Aggregation ◽  
Correlation Coefficient ◽  
Whole Blood ◽  
Blood Platelet ◽  
Inhibitory Effect ◽  
Time Interval ◽  
Adp Receptor ◽  
Spontaneous Platelet Aggregation

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

Mechanism of the Antiplatelet Action of Dipyridamole in Whole Blood: Modulation of Adenosine Concentration and Activity

1986 ◽  
Vol 55 (01) ◽  
pp. 012-018 ◽  
Author(s):  
Paolo Gresele ◽  
Jef Arnout ◽  
Hans Deckmyn ◽  
Jos Vermylen
Keyword(s):  
Platelet Aggregation ◽  
Adenosine Receptor ◽  
Whole Blood ◽  
Blood Cells ◽  
Inhibitory Action ◽  
Phosphodiesterase Inhibitor ◽  
Inhibitory Effect ◽  
Adenosine Concentration ◽  
Pharmacological Studies

SummaryDipyridamole inhibits platelet aggregation in whole blood at lower concentrations than in plasma. The blood cells responsible for increased effectiveness in blood are the erythrocytes. Using the impedance aggregometer we have carried out a series of pharmacological studies in vitro to elucidate the mechanism of action of dipyridamole in whole blood. Adenosine deaminase, an enzyme breaking down adenosine, reverses the inhibitory action of dipyridamole. Two different adenosine receptor antagonists, 5’-deoxy-5’-methylthioadenosine and theophylline, also partially neutralize the activity of dipyridamole in blood. Enprofylline, a phosphodiesterase inhibitor with almost no adenosine receptor antagonistic properties, potentiates the inhibition of platelet aggregation by dipyridamole. An inhibitory effect similar to that of dipyridamole can be obtained combining a pure adenosine uptake inhibitor (RE 102 BS) with a pure phosphodiesterase inhibitor (MX-MB 82 or enprofylline). Mixing the blood during preincubation with dipyridamole increases the degree of inhibition. Lowering the haematocrit slightly reduces the effectiveness.Although we did not carry out direct measurements of adenosine levels, the results of our pharmacological studies clearly show that dipyridamole inhibits platelet aggregation in whole blood by blocking the reuptake of adenosine formed from precursors released by red blood cells following microtrauma. Its slight phosphodiesterase inhibitory action potentiates the effects of adenosine on platelets.

scholarly journals Cathepsin L plays a key role in SARS-CoV-2 infection in humans and humanized mice and is a promising target for new drug development

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Miao-Miao Zhao ◽  
Wei-Li Yang ◽  
Fang-Yuan Yang ◽  
Li Zhang ◽  
Wei-Jin Huang ◽  
...  
Keyword(s):  
Drug Development ◽  
Cathepsin L ◽  
Human Cells ◽  
New Drugs ◽  
Disease Course ◽  
Promising Target ◽  
First Time

AbstractTo discover new drugs to combat COVID-19, an understanding of the molecular basis of SARS-CoV-2 infection is urgently needed. Here, for the first time, we report the crucial role of cathepsin L (CTSL) in patients with COVID-19. The circulating level of CTSL was elevated after SARS-CoV-2 infection and was positively correlated with disease course and severity. Correspondingly, SARS-CoV-2 pseudovirus infection increased CTSL expression in human cells in vitro and human ACE2 transgenic mice in vivo, while CTSL overexpression, in turn, enhanced pseudovirus infection in human cells. CTSL functionally cleaved the SARS-CoV-2 spike protein and enhanced virus entry, as evidenced by CTSL overexpression and knockdown in vitro and application of CTSL inhibitor drugs in vivo. Furthermore, amantadine, a licensed anti-influenza drug, significantly inhibited CTSL activity after SARS-CoV-2 pseudovirus infection and prevented infection both in vitro and in vivo. Therefore, CTSL is a promising target for new anti-COVID-19 drug development.

scholarly journals ROLE OF NITRIC OXIDE (NO) IN CAPSAICIN MEDIATED ANTI-PLATELET ACTIVITY IN IN VITRO, IN VIVO, EX-VIVO MODEL OF PLATELET AGGREGATION ASSAY AND ARTERIAL THROMBOSIS IN RAT: POTENTIAL THERAPEUTIC TARGET?

2018 ◽  
Vol 10 (4) ◽  
pp. 44
Author(s):  
Mihir K Patel ◽  
Kiranj K. Chaudagar ◽  
Anita A. Mehta
Keyword(s):  
Platelet Aggregation ◽  
Arterial Thrombosis ◽  
Ex Vivo ◽  
Platelet Activity ◽  
Aggregation Assay ◽  
Thrombosis Model ◽  
Platelet Aggregation Assay

Objective: Although recent advances in the treatment of congestive heart disease, mortality among patients’ remains a questionable remark. Therefore, we evaluated the role of capsaicin on in vitro and ex vivo platelet aggregation induced by Adenosine Di-Phosphate (ADP) as well as in in vivo thrombosis models and role of NO, KATP was also identified in the capsaicin-induced anti-platelet animal model as well as in vivo model of arterial thrombosis.Methods: According to body weight wistar rats were divided into five groups. Group I and Group II was treated with saline and capsaicin (3 mg/kg, i. v), while animals from Group III were treated with N(ω)-nitro-L-arginine methyl ester (L-NAME) (30 mg/kg, i. v) 30 min before administration of capsaicin (3 mg/kg, i. v). Group IV animals were treated with glibenclamide (10 mg/kg,i. v) 30 min before administration of capsaicin (3 mg/kg, i. v). Group V was considered as a positive control and administered clopidogrel (30 mg/kg, p. o). Animals were subjected for in vitro, ex-vivo platelet aggregation assay. ADP (30µM) was utilized as an aggregating agent in these experiments. After these assays; animals of each group were subjected for subaqueous tail bleeding time in a rat model and FeCl3-induced arterial thrombosis model in rats.Results: In ADP-induced in vitro platelet aggregation, a significant reduction in % platelet aggregation was observed at 50µM (64.35±4.641) and 100µM (52.72±4.192) concentration of capsaicin as compared to vehicle control (85.82±3.716). Capsaicin (3 mg/kg, i. v) also showed a significant reduction (49.53±4.075) in ex-vivo ADP-induced platelet aggregation as compared to vehicle control (89.38±2.057). In FeCl3 induced arterial thrombosis model, Capsaicin (3 mg/kg, i. v) exhibited an increase in time to occlusion in this rodent model and presence of the L-NAME and glibenclamide had inhibited the activity of capsaicin.Conclusion: In our study, capsaicin (50 µM, 100µM) exhibited potent anti-platelet activity in ADP-induced platelet aggregation, similarly capsaicin exhibited significant anti-platelet action in the ex-vivo study. Moreover, the presence of L-NAME and glibenclamide inhibited the anti-thrombotic and anti-platelet action of capsaicin. Therefore, it was concluded that NO and KATP may be involved in the anti-thrombotic action of capsaicin.

EFFECTS OF BLOOD CELLS ON PLATELET AGGREGATION BY IMPEDANCE METHOD

1987 ◽  
Author(s):  
L Mannucci ◽  
R Redaelli ◽  
E Tremoll
Keyword(s):  
Platelet Aggregation ◽  
Whole Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Normal Subjects ◽  
Impedance Method ◽  
Induced Aggregation ◽  
Vitro Data ◽  
In Vitro Data

To evaluate the effects of blood cells on the response of platelets to aggregating agents using whole blood impedance aggregometer, studies were carried out on whole blood (WB) of normal subjects and of patients with: polycythemia vera (PV), iatrogenic anemia (IA), primary thrombocytosis (PT), idiopathic thrombotic purpura (ITP), myeloid chronic leukemia (MCL), iatrogenic leukopenia (IL). The in vitro effects of red blood cells (RBC) and of white blood cells (WBC) on platelet rich plasma (PRP) aggregation were also evaluated. WB, PRP, WBC and RBC were prepared by conventional methods. Aggregation was performed using the impedance aggregometer (mod. 540, Chrono Log Corp). In normal subjects the concentration of collagen giving 50 % aggregation (AC50 ) found in PRP did not differ from that of WB, indicating that hematocrit values within the normal range did not appreciably affect platelet aggregation. The results obtained in WB of patients are summarized in the table: In vitro data showed that aggregation in prp in wb of normal subjects was related to the number of platelets present in the sample. RBC added to PRP significant reduced aggregation only when the RBC number was greater than 4.101 cells. No effect of WBC on collagen induced aggregation of PRP was observed, whereas significant inhibition was detected after ADP. It is concluded that the aggregation evaluated in WB with impedance method is dependent on the platelet number. Also, in vitro data and studies in WB of patients indicate that aggregation is significantly affected by the presence of cells other than platelets only in conditions of changes of the ratio between platelets and leukocytes and/or red cells.

scholarly journals Antagonists of the system L neutral amino acid transporter (LAT) promote endothelial adhesivity of human red blood cells

2017 ◽  
Vol 117 (07) ◽  
pp. 1402-1411 ◽  
Author(s):  
Laura Beth Mann Dosier ◽  
Vikram J. Premkumar ◽  
Hongmei Zhu ◽  
Izzet Akosman ◽  
Michael F. Wempe ◽  
...  
Keyword(s):  
Amino Acid ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Amino Acid Transporter ◽  
Neutral Amino Acid ◽  
Neutral Amino Acid Transporter ◽  
System L

SummaryThe system L neutral amino acid transporter (LAT; LAT1, LAT2, LAT3, or LAT4) has multiple functions in human biology, including the cellular import of S-nitrosothiols (SNOs), biologically active derivatives of nitric oxide (NO). SNO formation by haemoglobin within red blood cells (RBC) has been studied, but the conduit whereby a SNO leaves the RBC remains unidentified. Here we hypothesised that SNO export by RBCs may also depend on LAT activity, and investigated the role of RBC LAT in modulating SNO-sensitive RBC-endothelial cell (EC) adhesion. We used multiple pharmacologic inhibitors of LAT in vitro and in vivo to test the role of LAT in SNO export from RBCs and in thereby modulating RBC-EC adhesion. Inhibition of human RBC LAT by type-1-specific or nonspecific LAT antagonists increased RBC-endothelial adhesivity in vitro, and LAT inhibitors tended to increase post-transfusion RBC sequestration in the lung and decreased oxygenation in vivo. A LAT1-specific inhibitor attenuated SNO export from RBCs, and we demonstrated LAT1 in RBC membranes and LAT1 mRNA in reticulocytes. The proadhesive effects of inhibiting LAT1 could be overcome by supplemental L-CSNO (S-nitroso-L-cysteine), but not D-CSNO or L-Cys, and suggest a basal anti-adhesive role for stereospecific intercellular SNO transport. This study reveals for the first time a novel role of LAT1 in the export of SNOs from RBCs to prevent their adhesion to ECs. The findings have implications for the mechanisms of intercellular SNO signalling, and for thrombosis, sickle cell disease, and post-storage RBC transfusion, when RBC adhesivity is increased.

scholarly journals Haemodynamic flow conditions at the initiation of high-shear platelet aggregation: a combined in vitro and cellular in silico study

2020 ◽  
Vol 11 (1) ◽  
pp. 20190126 ◽  
Author(s):  
B. J. M. van Rooij ◽  
G. Závodszky ◽  
A. G. Hoekstra ◽  
D. N. Ku
Keyword(s):  
Platelet Aggregation ◽  
In Silico ◽  
Platelet Rich Plasma ◽  
High Shear ◽  
Shear Rates ◽  
Flow Simulations ◽  
In Silico Study ◽  
Platelet Aggregates

The influence of the flow environment on platelet aggregation is not fully understood in high-shear thrombosis. The objective of this study is to investigate the role of a high shear rate in initial platelet aggregation. The haemodynamic conditions in a microfluidic device are studied using cell-based blood flow simulations. The results are compared with in vitro platelet aggregation experiments performed with porcine whole blood (WB) and platelet-rich-plasma (PRP). We studied whether the cell-depleted layer in combination with high shear and high platelet flux can account for the distribution of platelet aggregates. High platelet fluxes at the wall were found in silico . In WB, the platelet flux was about twice as high as in PRP. Additionally, initial platelet aggregation and occlusion were observed in vitro in the stenotic region. In PRP, the position of the occlusive thrombus was located more downstream than in WB. Furthermore, the shear rates and stresses in cell-based and continuum simulations were studied. We found that a continuum simulation is a good approximation for PRP. For WB, it cannot predict the correct values near the wall.

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