Activities of the enzymes NTPDase, 5'-nucleotidase and adenosine deaminase in platelets of rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae
Tonin, Alexandre AlbertoAzevedo, Maria Isabel deSilva, Aleksandro Schafer daCosta, Marcio MachadoFrança, Raqueli TeresinhaPimentel, Victor CâmeraSouza, Viviane do Carmo GonçalvesJaques, Jeandre Augusto dos SantosLeal, Claudio Alberto MartinsBadke, Manoel Renato TelesLeal, Daniela Bitencourt RosaSchetinger, Maria Rosa ChitolinaLopes, Sonia Terezinha dos Anjos
Background: Leptospirosis, a spirochetal zoonotic disease caused by different serovars of Leptospira interrogans, is increasingly recognized as an important cause of hemorrhagic fever. Although the haemorrhagic potential of leptospirosis was noted by Weil (1886) as early as 1886, its pathophysiology is still not clearly elucidated, particularly regarding the cause and mechanisms of bleeding. Studies with ectonucleoside triphosphate diphosphohydrolase (NTPDase; EC 3.6.1.5; CD39), 5'-nucleotidase (EC 3.1.3.5; CD73) and adenosine deaminase (ADA) have demonstrated the involvement of these enzymes in thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. Since leptospirosis is a disease increasingly recognized as an important cause of hemorrhagic fever, the aim of this study was to evaluate these enzymes activities and parameters of platelet aggregation in platelets from rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae during different periods of experimental infection. Materials, Methods & Results: For this purpose, thirty-six adult male rats were divided into two groups: A, as uninfected control (subgroups A1, A2 and A3); and B, infected (subgroups B1, B2 and B3). Group B was inoculated intraperitoneally (Day 0) with 2 x 108 organisms per rat. Blood samples were collected on days 05 (A1 and B1), 10 (A2 and B2) and 20 (A3 and B3) post-inoculation (PI). In the infected group, platelet count had a decrease on day 10 PI and prothrombin time (PT) had an increase on day 5 PI. In the same group, platelet aggregation decreased (P < 0.01) day 10 PI. The hydrolysis of ATP in platelets was also decreased (P < 0.05) on day 10 PI, when compared to the control group. By the other side, ADP hydrolysis was increased (P < 0.05) on days 5 and 10 PI. 5'-nucleotidase activity was significantly increased on day 5 (P < 0.01) and 20 (P < 0.05) PI. Results of adenosine deamination into inosine by ADA in platelets showed a signifi cant (P < 0.01) increase on days 5 and 10 PI in the infected group. Discussion: Studies with NTPDase, 5'nucleotidase and ADA have demonstrated the involvement of these enzymes in the thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. It has been established that extracellular adenosine nucleotides and adenosine are versatile signaling molecules known to participate in an array of platelet functions. For example, the nucleotide ADP is the main promoter of platelet aggregation, while adenosine can act as a vasodilator and an inhibitor of platelet aggregation. In addition, high concentrations of ATP have been shown to inhibit ADP-induced aggregation in vitro, while low concentrations of ATP can significantly enhance platelet aggregation. In our experimental study the coagulation cascade was activated, since when the activities of NTPDase, 5'-nucleotidase and ADA were analyzed is possible to suggest that levels of ATP were decreased, unlike of ADP and AMP levels, supposedly increased during determinate periods of our experiment. Adenosine levels were also enhanced due to the higher levels of its precursors. This cascade activation may be a mechanism of bleeding prevention front to leptospires infection, especially the ones caused by serovar icterohaemorrhagiae.
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