Análise do potencial inibitório do ácido clorogênico e do ácido rosmarínico frente aos efeitos da peçonha da serpente Bothrops Leucurus Wagler (1824)

Abstract

Snakebites represent a serious public health problem in tropical countries. Bothrops leucurus snake is one of the main responsible for snakebites in Northeast Brazil, but this species is not included in the pool used in the production of antiophidic serum. Considering the limitations of the antiophidic serum, several studies have been carried out in order to develop complementary treatments for snakebite and, in this context, plants are an important source of bioactive compounds. Chlorogenic acid and rosmarinic acid are molecules derived from the secondary metabolism of plants, with several biological activities described and antiophidic potential still little explored. Thus, the aim of the study was to evaluate the inhibitory potential of chlorogenic and rosmarinic acids against the effects of B. leucurus venom. Initially, in vitro neutralization capacity of the enzymatic activities of the venom was evaluated. Then, in vivo tests were performed to verify the inhibition of acids against hemorrhagic, edematogenic, myotoxic activities and quantification of inflammatory markers, as well as the analysis of biochemical, hemostatic, hematological markers, and verification of the degree of lipid peroxidation in renal and liver tissues. Finally, the ability of the inhibitors to interact in silico with two classes of toxins was evaluated. The results showed that chlorogenic and rosmarinic acids were effective in inhibiting phospholipase A2 (21% and 60%), proteolytic (39% and 60%), hyaluronidase (59% and 75%) and coagulant (p < 0.0001) activities, respectively, as well as were effective in preventing fibrinogen degradation. In in vivo assays, the inhibitors attenuated edema (p < 0.01), reducing the levels of myeloperoxidase (p < 0.001), IL-6 (p < 0.0001) and IL-1β (p < 0.001) interleukins. The compounds also inhibited the hemorrhagic (p < 0.0001) and myotoxic (p < 0.001 – chlorogenic acid, p < 0.0001 – rosmarinic acid) actions of the venom. In serum biochemical markers, the inhibitors attenuated changes in urea levels (p < 0.01 - chlorogenic acid and p < 0.05 - rosmarinic acid), creatinine (p < 0.0001), uric acid (p < 0.0001 ), alanine aminotransferase (p < 0.05), aspartate aminotransferase (p < 0.001- chlorogenic acid and p < 0.05 - rosmarinic acid), amylase (p < 0.01), total proteins (p < 0.01), creatine kinase (p < 0.01) and lactate dehydrogenase (p < 0.0001). Furthermore, both compounds decreased the levels of lipid peroxidation in liver (p < 0.001) and renal (p < 0.0001) tissues. When evaluating hemostatic parameters, only rosmarinic acid was able to reduce the effects on circulating fibrinogen (p < 0.0001) and platelet count (p < 0.0001). Computational studies indicated that the inhibitors have the ability to interact with toxins of the phospholipase A2 and metalloprotease class. Taken together, the results obtained evidenced the antiophidic potential of chlorogenic and rosmarinic acids, which presented, in general, greater inhibition efficiency than the antiophidic serum. Thus, both inhibitors are promising candidates for the development of future adjuvants to be used to complement the currently available sorotherapy.