Influência do estabilizante sacarose e liofilizados contendo antiveneno: avaliação renal in vitro e in vivo

Abstract

Accidents involving snakes are a serious public health problem, often neglected, that affects the entire world. This type of condition is associated with a large number of morbidity and mortality when not treated in time. Antivenom or serum therapy is the only indication for the treatment of these accidents. It consists of the parenteral use of specific immunoglobulins that neutralize the circulating toxins of the venom. Currently, antivenoms can be presented in liquid or lyophilized form. Freeze-dried forms can be stored at temperatures above 25°C and are recommended and distributed to regions where cold storage cannot be guaranteed (tropical or desert regions of the world). During the freeze-drying process, stabilizers and/or lyoprotectants are added to the formulation that protect proteins from the thermal stress of the process. One of them is sucrose, which is associated, when administered intravenously, with the induction of acute kidney injuries. Thus, this study aimed to evaluate the renal effects caused by a lyophilized polyspecific antivenom formulated with two different concentrations of sucrose. For this, two models were used: in vitro, which uses renal proximal tubule cells, belonging to the LLC-MK2 lineage, and in vivo, which uses female Swiss mice, which were injected intravenously with specific doses of antivenom. As results of in vitro tests, changes in cell viability, changes in oxidative stress markers (in both models studied) and morphological changes in renal architecture were observed, similar to osmotic nephrosis induced by exogenous solutes. Proteomic mapping of kidney tissue was also carried out to observe the proteins involved in the changes found. On the other hand, antivenoms, even showing signs of alteration in renal homeostasis, possibly associated with the presence of sucrose, were able to neutralize the action of the total venom of the Bothrops asper snake in the in vitro model. The action of the isolated antivenin in the in vivo model showed results similar to those found in in vitro tests. However, the formulation with 5% sucrose showed an action similar to that of the antivenom formulated without sucrose and without much associated damage. As future perspectives, a study is needed to evaluate the joint action of poison and antivenom, in order to complement and clarify the results obtained so far, thus attesting to the safety and effectiveness of antivenom in the regression of poisoning symptoms.