V. Chernyshenko¹, E. Snezhkova², M. Mazur², T. Chernyshenko¹,
T. Platonova¹, O. Sydorenko², E. Lugovskoy¹, V. Nikolaev²

¹Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: bio.cherv@gmail.com;
²RE Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine, Kyiv

Received: 13 December 2018; Accepted: 20 March 2019

Radiation-induced coagulopathy (RIC) is one of the major causes of death during acute radiation syndrome (ARS). The aim of this study was to characterize the responses of the hemostasis system to ARS of a moderate level on the 1st and 9th days after irradiation. We aimed to identify molecular markers of the blood coagulation system that are most affected by ARS and to estimate the enterosorption effect on the development of irradiation-induced changes. Platelet aggregation rate, activated partial thromboplastin time (APTT) and fibrinogen concentration were determined by standard methods. Level of protein C (PC) was measured using­ chromogenic substrate S2366 (p-Glu-Pro-Arg-pNa) and Agkistrodon halys halys snake venom activa­ting enzyme. Functionally inactive forms of prothrombin (FIFPs) were determined using two activators in parallel – thromboplastin or prothrombin activator from Echis multisqumatis venom. Rats of both irradia­ted groups had a higher risk of intravascular clotting in comparison to both control groups. Statistically significant shortening of clotting time in the APTT test (24 ± 4 s vs. 33 ± 5 s) and increased fibrinogen concentration (4.2 ± 0.6 mg/ml vs. 3.2 ± 0.3 mg/ml) were detected. Both parameters were normalized on the 9th day after irradiation. However the platelet count was decreased (0.3∙10⁶ ± 0.05∙10⁶ 1/μl vs. 0.145∙10⁶ ± 0.04∙10⁶ 1/μl) due to the impaired megakaryocytic function. The level of PC was decreased after X-ray irradiation (70 ± 10%) and partly restored on the 9th day after irradiation (87 ± 10%). Administration of activated carbon (AC) inhibited the drop in the PC concentration after X-ray irradiation (86 ± 15%) and accelerated its restoration on the 9th day (103 ± 14%). The statistically significant accumulation of FIFPs was detected in blood plasma of irradia­ted rats at the 1st and 9th days after irradiation. No FIFPs were found in any irradiated rat treated with AC. Characterization of the hemostasis system of rats that were exposed to a semilethal dose of X-rays allowed us to select parameters that can be used for monitoring of ARS development. Apart of from basic coagulation tests (APTT) and the measurement of platelet aggregation, fibrinogen and protein C level we can recommend the determination of FIFPs as a useful tool for estimation of the hemostasis response after irradiation with X-rays. This test indicates the intravascular thrombin generation and can help predict thrombotic complication or disseminated intravascular coagulation. Determination of FIFPs in blood plasma of irradia­ted rats allowed us to study the enterosorption effect on the development of irradiation-induced changes. It was shown that enterosorption with AC prevented accumulation of FIFPs which appears to be a newly discovered anti-thrombotic effect of therapy with AC. ARS influenced hemostasis by inducing thrombin generation (indicated by FIFPs generation), low-grade inflammation (indicated by PC concentration decrease) and thrombocytopenia. Enterosorption with AC minimizes inflammation and pro-coagulant processes caused by a moderate dose of X-ray irradiation. Accumulation of FIFPs can be assumed to be one of the most sensitive markers of the blood coagulation response to X-ray irradiation.

Keywords: activated carbon, acute radiation syndrome of moderate level, blood coagulation, enterosorption, platelets