The Effects of Restraint and Cold Restraint Stress on Coagulation Indicators in Wistar Albino Rats

Yıl 2023, , 160 - 165, 30.06.2023

Çağlasu Koç , Mehmet Ekici

https://doi.org/10.30607/kvj.1239878

Öz

The aim of this study was to investigate the effects of restraint and cold restraint stress from acute stress protocols on coagulation indicators in rats. The study was conducted in 18 male Wistar albino rats aged 8-10 weeks with a body weight of 180-220 g. After a one-week adaptation period, the rats were randomly divided into three groups (n=6/group). The animals in the control group were not exposed to any stress. Rats in the restraint group were housed in restrainers designed for rats, and their movement was restricted for 2 hours at room temperature. Rats in the cold restraint group were kept in the restrainer at +4°C for 2 hours. Blood samples were collected by cardiac puncture under ketamine and xylazine anesthesia in Vacutainer® tubes containing 3.2% sodium citrate (0.109 M trisodium citrate). Coagulation indicators (aPTT, PT, INR, fibrinogen, and D-dimer) were analyzed using an automated analyzer (Roche Cobas t511, Switzerland). Although acute stress (restraint and cold restraint stress) had no effect on aPTT and D-dimer levels (p>0.05), it increased PT and INR values (p<0.05) and decreased fibrinogen concentration (p<0.05). Consequently, acute stress may lead to platelet hypofunction in rats by prolonging PT, increasing INR, and decreasing fibrinogen concentration.

Anahtar Kelimeler

Acute stress, aPTT, D-dimer, Fibrinogen, INR, PT

Destekleyen Kurum

TUBİTAK

Proje Numarası

2209- A - Üniversite Öğrencileri Araştırma Projeleri

Kısıtlama ve Soğuk Kısıtlama Stresinin Wistar Albino Sıçanlarda Pıhtılaşma Göstergeleri Üzerindeki Etkileri

Öz

Bu çalışmanın amacı, kısıtlama ve soğuk kısıtlama stresi gibi iki farklı akut stres protokolünün sıçanlarda pıhtılaşma göstergeleri üzerindeki etkilerini araştırmaktı. Çalışma, vücut ağırlığı 180-220 g olan, 8-10 haftalık 18 erkek Wistar albino sıçan üzerinde gerçekleştirildi. Bir haftalık adaptasyon sürecinden sonra sıçanlar rastgele üç gruba (n=6/grup) ayrıldı. Kontrol grubundaki hayvanlar herhangi bir strese maruz bırakılmadı. Kısıtlama grubundaki ratlar, ratlar için tasarlanmış kısıtlayıcı içinde barındırıldı ve oda sıcaklığında 2 saat hareketleri kısıtlandı. Soğuk kısıtlama grubundaki ratlar +4°C'de 2 saat kısıtlayıcı içinde tutuldu. Kan numuneleri, %3,2 sodyum sitrat (0.109 M trisodyum sitrat) içeren Vacutainer® tüplerine ketamin ve ksilazin anestezisi altında kardiyak punksiyon yoluyla toplandı. Pıhtılaşma göstergeleri (aPTT, PT, INR, fibrinojen ve D-dimer) otomatik bir analiz cihazı (Roche Cobas t511, İsviçre) kullanılarak analiz edildi. Akut stres (kısıtlama ve soğuk kısıtlama stresi) aPTT ve D-dimer düzeylerine etki etmezken (p>0.05), PT ve INR değerlerini artırdı (p<0.05), fibrinojen konsantrasyonunu düşürdü (p<0.05). Sonuç olarak akut stres, PT'yi uzatarak, INR'yi artırarak ve fibrinojen konsantrasyonunu azaltarak sıçanlarda trombosit hipofonksiyonuna yol açabilir.

Anahtar Kelimeler

Akut stres, aPTT, D-dimer, Fibrinojen, INR, PT

Proje Numarası

2209- A - Üniversite Öğrencileri Araştırma Projeleri

Kaynakça

• Austin AW, Wirtz PH, Patterson SM, Stutz M, von Känel R. Stress-induced alterations in coagulation: Assessment of a new hemoconcentration correction technique. Psychosom Med. 2012;74(3).
• de Boer D, Ring C, Wood M, Ford C, Jessney N, McIntyre D, Carroll D. Time course and mechanisms of mental stress-induced changes and their recovery: Hematocrit, colloid osmotic pressure, whole blood viscosity, coagulation times, and hemodynamic activity. Psychophysiology. 2007;44(4).
• Bondarchuk YA, Alekseeva O v., Shakhmatov II, Krinitsina IN. Response of the Hemostatic System to Acute Vital Stress in Rats. Bull Exp Biol Med. 2020;168(6). García-Manzano A, González-Llaven J, Lemini C, Rubio-Póo C. Standardization of rat blood clotting tests with reagents used for humans. Proc West Pharmacol Soc. 2001.
• Hata T, Kawabata A, Itoh E. Platelet hypoaggregability in rats exposed to SART stress (repeated cold stress). Thromb Res. 1992;65(4–5).
• Hatu T, Kawabata A, Itoh E. Blood Coagulation and Fibrinolysis in SART-Stressed (Repeated Cold-Stressed) Rats and Drug Effects on the Altered Hemostatic Parameters. The Japanese Journal of Pharmacology. 1991;56(4).
• Kamarck TW, Debski TT, Manuck SB. Enhancing the laboratory-to-life generalizability of cardiovascular reactivity using multiple occasions of measurement. Psychophysiology. 2000;37(4).
• von Känel R, Dimsdale JE. Effects of sympathetic activation by adrenergic infusions on hemostasis in vivo. Eur J Haematol. 2000.
• von Känel R, Kudielka BM, Haeberli A, Stutz M, Fischer JE, Patterson SM. Prothrombotic changes with acute psychological stress: Combined effect of hemoconcentration and genuine coagulation activation. Thromb Res. 2009;123(4).
• von Känel R, Mills PJ, Fainman C, Dimsdale JE. Effects of psychological stress and psychiatric disorders on blood coagulation and fibrinolysis: A biobehavioral pathway to coronary artery disease? Psychosom Med. 2001;63(4).
• von Känel R, Preckel D, Zgraggen L, Mischler K, Kudielka BM, Haeberli A, Fischer JE. The effect of natural habituation on coagulation responses to acute mental stress and recovery in men. Thromb Haemost. 2004;92(6).
• Kawabata A, Hata T. Characterization of platelet hypofunctions in rats under SART stress (repeated cold stress). Thromb Res. 1993;69(2).
• Kitchen S, Geisen U, Kappelmayer J, Quehenberger P, Lowe A, Jones R, Miles G, Boehm JG, Rozsnyai G. Evaluating the analytical performance of four new coagulation assays for the measurement of fibrinogen, D-dimer and thrombin time. Int J Lab Hematol. 2018a;40(6).
• Kitchen S, Geisen U, Kappelmayer J, Quehenberger P, Drieß J, Lowe A, Jones R, Boehm JG, Miles G, Rozsnyai G. Evaluating the analytical performance of five new coagulation assays for the measurement of prothrombin time and activated thromboplastin time. Int J Lab Hematol. 2018b Dec;40(6).
• Lewis JH, van Thiel DH, Hasiba U, Spero JA, Gavaler J. Comparative hematology and coagulation: Studies on rodentia (Rats). Comp Biochem Physiol A Physiol. 1985;82(1).
• Loban’-Chereda GA, Novosel’tseva T v. [Coagulative capacity of the blood and anti-aggregative activity of the vascular wall in rats subjected to immobilization stress]. Fiziol Zh [Internet]. 1990;36(2): 13—18. Available from: http://europepmc.org/abstract/MED/2113873
• Malyszko J, Urano T, Takada Y, Takada A. Time-dependent changes in platelet aggregation, fibrinolytic activity, and peripheral serotonergic measures in rats subjected to water immersion restraint stress. Haemostasis. 1994;24(4).
• Matsuhisa F, Kitamura N, Satoh E. Effects of acute and chronic psychological stress on platelet aggregation in mice. Stress. 2014;17(2).
• Pethaperumal S, Hung SC, Lien TS, Sun DS, Chang HH. P-Selectin is a Critical Factor for Platelet-Mediated Protection on Restraint Stress-Induced Gastrointestinal Injury in Mice. Int J Mol Sci. MDPI; 2022;23(19).
• Sandrini L, Ieraci A, Amadio P, Popoli M, Tremoli E, Barbieri SS. Apocynin Prevents Abnormal Megakaryopoiesis and Platelet Activation Induced by Chronic Stress. Oxid Med Cell Longev. 2017;2017.
• Sandrini L, Ieraci A, Amadio P, Zarà M, Barbieri SS. Impact of acute and chronic stress on thrombosis in healthy individuals and cardiovascular disease patients. Int J Mol Sci. 2020.
• Sapolsky RM, Krey LC, McEwen BS. The adrenocortical stress-response in the aged male rat: impairment of recovery from stress. Exp Gerontol. 1983;18(1).
• Sapolsky RM, Krey LC, McEwen BS. The neuroendocrinology of stress and aging: the glucocorticoid cascade hypothesis. Endocr Rev. 1986; 7(3).
• Strike PC, Magid K, Brydon L, Edwards S, McEwan JR, Steptoe A. Exaggerated platelet and hemodynamic reactivity to mental stress in men with coronary artery disease. Psychosom Med. 2004;66(4).
• Takeda H, Asaka M, Matsuno K, Ohtaki T, Miyazaki T. Stress-induced gastric mucosal lesion and platelet aggregation in rats. J Clin Gastroenterol. 1992;14.
• Tu BX, Wang LF, Zhong XL, Hu ZL, Cao WY, Cui YH, Li SJ, Zou GJ, Liu Y, Zhou SF, Zhang WJ, Su JZ, Yan XX, Li F, Li CQ. Acute restraint stress alters food-foraging behavior in rats: Taking the easier Way while suffered. Brain Res Bull. 2019;149.
• Umarova BA, Shapiro FB, Kogan AE, Strukova SM. Mast cells secrete heparin during immobilization stress in rats. Thromb Res. 1997;85(3).
• Wirtz PH, Redwine LS, Baertschi C, Spillmann M, Ehlert U, von Känel AR. Coagulation activity before and after acute psychosocial stress increases with age. Psychosom Med. 2008;70(4).
• Yoshihara H, Yamamoto T, Mihara H. Changes in coagulation and fibrinolysis occurring in dogs during hypothermia. Thromb Res. 1985;37(4).
• Zgraggen L, Fischer JE, Mischler K, Preckel D, Kudielka BM, von Känel R. Relationship between hemoconcentration and blood coagulation responses to acute mental stress. Thromb Res. 2005;115(3).
• Zhu D, Tong Q, Liu W, Tian M, Xie W, Ji L, Shi J. Angiotensin (1-7) protects against stress-induced gastric lesions in rats. Biochem Pharmacol. 2014;87(3).