A simple and efficient method for platelet isolation from their plasma

Lagarde, Michel; Bryon, Paul‐André; Guichardant, Michel; Dechavanne, M

doi:10.1016/0049-3848(80)90098-5

Cited by 178 publications

(49 citation statements)

References 8 publications

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“…Blood samples were collected into tubes containing acid-citrate-dextrose (ACD; 0.8% citric acid, 2.2% sodium citrate, and 2.45% dextrose; 6:1 vol/vol). Platelet-rich plasma was prepared as previously described (13). Briefly, platelet-rich plasma obtained by centrifugation of the blood at 100g for 15 min was acidified to pH 6.4 with citric acid and centrifuged at 900g for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

et al. 2004

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Vascular complications are the leading causes of morbidity and mortality in diabetic patients. The contribution of platelets to thromboembolic complications is well documented, but their involvement in the initiation of the atherosclerotic process is of rising interest. Thus, the aim of the present study was to evaluate basal arachidonic acid metabolism in relation to the redox status of platelets in both type 1 and type 2 diabetic patients, in the absence of vascular complications, as compared with respective control subjects. For the first time, we show that basal thromboxane B 2 , the stable catabolite of thromboxane A 2 , significantly increased in resting platelets from both type 1 and type 2 diabetic patients (58 and 88%, respectively), whereas platelet malondialdehyde level was only higher in platelets from type 2 diabetic subjects (67%). On the other hand, both vitamin E levels and cytosolic glutathione peroxidase activities were significantly lower in platelets from diabetic patients as compared with respective control subjects. We conclude that platelet hyperactivation was detectable in well-controlled diabetic patients without complications. This abnormality was associated with increased oxidative stress and impaired antioxidant defense in particular in type 2 diabetic patients. These alterations contribute to the increased risk for occurrence of vascular diseases in such patients. Diabetes

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Section: Methodsmentioning

confidence: 99%

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

et al. 2004

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“…Platelets were isolated [26] and incubated for 2 h at 37°C in the presence or absence of LDL (1 mg/ml).…”

Section: Patients and Control Volunteersmentioning

confidence: 99%

LDL from obese patients with the metabolic syndrome show increased lipid peroxidation and activate platelets

et al. 2011

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Aims/hypothesis This study assessed oxidative stress in LDL from obese patients with the metabolic syndrome and compared it with that in LDL from type 2 diabetic patients or control volunteers. It also determined the effect on platelets of LDL from the three groups. Methods The profiles of lipids, fatty acids and fatty acid oxidation products were determined in LDL isolated from plasma of patients with the metabolic syndrome, patients with type 2 diabetes and volunteers (n=10 per group). The effects of LDL from the participant groups on the platelet arachidonic acid signalling cascade and aggregation were investigated. Results Compared with LDL from control volunteers, LDL from obese metabolic syndrome and type 2 diabetic patients had lower cholesteryl ester, higher triacylglycerol and lower ethanolamine plasmalogen levels. Proportions of linoleic acid were decreased in phosphatidylcholine and cholesteryl esters in LDL from both patient groups. Among the markers of lipid peroxidation, oxidation products of linoleic acid (hydroxy-octadecadienoic acids) and malondialdehyde were increased by 59% and twofold, respectively in LDL from metabolic syndrome and type 2 diabetic patients. LDL from metabolic syndrome and type 2 diabetic patients were equally potent in activating the platelet arachidonic acid signalling cascade through increased phosphorylation of p38 mitogen-activated protein kinase and cytosolic phospholipase A 2 , and through increased thromboxane B 2 formation. LDL from patients with the metabolic syndrome and type 2 diabetes potentiated platelet aggregation by threefold and 3.5-fold respectively, whereas control LDL had no activating effects on platelets. Conclusions/interpretation The metabolic syndrome in obese patients, without or with diabetes, is associated with increased oxidative stress in LDL, which triggers platelet activation.

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“…Blood was centrifuged at 200g for 15min at 20°C. The platelet-rich plasma (PRP) was removed and platelets were isolated as described previously (Lagarde et al, 1980). Briefly, the PRP was acidified to pH 6.4 with 0.15 M citric acid and centrifuged at 900g for 10min at 20°C in siliconized tubes.…”

Section: Cell Preparation Proceduresmentioning

confidence: 99%

Thrombin‐activated platelets promote leukotriene B₄ synthesis in polymorphonuclear leucocytes stimulated by physiological agonists

Palmantier

Borgeat

1991

British J Pharmacology

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The addition of 2 × 108 human platelets to 8 × 106 polymorphonuclear leucocytes (PMNL) incubated in presence of 2.5 u ml−1 thrombin and 0.1 μm N‐formyl‐Met‐Leu‐Phe (FMLP) (or C5a or PAF) led to enhancement of leukotriene B4 (LTB4) synthesis by the PMNL (measured by h.p.l.c. as 20‐hydroxy‐ and 20‐carboxy‐LTB4) from 4 ± 1 pmol (in absence of platelets) to 26 ± 4 pmol (mean ± s.e.mean, n = 9). Platelets and thrombin were both essential for the enhancement of LTB4 synthesis. Platelets also caused enhancement of LTB4 synthesis from (30 ± 12 to 134 ± 25 pmol, n = 6) when PMNL pretreated with granulocyte‐macrophage colony‐stimulating factor were used in similar experiments. Enhancement of LTB4 synthesis was also observed (from 5 ± 1.5 to 26.5 ± 5 pmol, n = 9) when the supernatants of thrombin‐activated platelet suspensions were added to FMLP‐stimulated PMNL. Supernatants of platelet suspensions activated by thrombin in presence of cyclo‐oxygenase and 12‐lipoxygenase inhibitors led to greater enhancement (from 5 ± 3 to 153.5 ± 27.5 pmol, n = 3) of LTB4 synthesis by FMLP‐stimulated PMNL, suggesting that arachidonic acid itself, rather than its metabolites was responsible for the effects of platelets. Addition of arachidonic acid to FMLP‐stimulated PMNL at a concentration comparable to that measured in thrombin‐activated platelet supernatants (0.2 ± 0.025 μm, n = 6) mimicked the effect of platelets or platelet supernatants on LTB4 synthesis in FMLP‐activated PMNL. The present data indicate that under conditions of cell activation by physiological agonists, platelets can significantly increase the formation of the proinflammatory compound LTB4 in PMNL by providing arachidonic acid. These data lend support to the concept that platelet‐PMNL interactions could modulate the inflammatory process.

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A simple and efficient method for platelet isolation from their plasma

Cited by 178 publications

References 8 publications

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

LDL from obese patients with the metabolic syndrome show increased lipid peroxidation and activate platelets

Thrombin‐activated platelets promote leukotriene B₄ synthesis in polymorphonuclear leucocytes stimulated by physiological agonists

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A simple and efficient method for platelet isolation from their plasma

Cited by 178 publications

References 8 publications

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

Diabetic Patients Without Vascular Complications Display Enhanced Basal Platelet Activation and Decreased Antioxidant Status

LDL from obese patients with the metabolic syndrome show increased lipid peroxidation and activate platelets

Thrombin‐activated platelets promote leukotriene B4 synthesis in polymorphonuclear leucocytes stimulated by physiological agonists

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Thrombin‐activated platelets promote leukotriene B₄ synthesis in polymorphonuclear leucocytes stimulated by physiological agonists