Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Abstract

BackgroundGlobally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes.MethodsHuman erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0–640 μg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination.ResultsLow concentrations of venom (<10 μg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 μg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 μg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm−1) and lipid peroxidation (2960, 2920 and 1740 cm−1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom.Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.

The Urgent Need to Develop Novel Strategies for the Diagnosis and Treatment of Snakebites

“…This produces an α-keto acid, ammonia and hydrogen peroxide, all of which can have cytotoxic effects. The hydrogen peroxide produced may additionally lead to the oxidation of haemoglobin seen as a result of some viper venoms [84,85]. They may also induce oedema [86] and apoptosis [87], as well as acting as anti-coagulants via the inhibition of factor IX [88].…”

Impact of Naja nigricollis Venom on the Production of Methaemoglobin

“…The ability of venoms to oxidise Hb into MetHb has been reported within the family of Viperidae [15,16,17,26]. Here, we screened 47 different viper (30) and elapid (17) venoms from the targeted venom discovery array for the cardiovascular system (T-VDA CV ) in order to determine the impact of these on MetHb production.…”

“…A previous study using Crotalus molossus nigrescens venom reported that high concentrations of this venom cause some level of proteolytic degradation to Hb [26]. The same study postulated that the venom’s conversion of Hb to MetHb is due to the oxidative stress induced by H 2 O 2 which is produced by venom LAAO [26]. It may be possible that following this, H 2 O 2 then reacts with the ferrous Hb producing further reactive species via the Fenton reaction [31].…”

Evaluation of Thiol-dependent Enzymes on the Pharmacological Effects Induced by the Catalytically Active PLA2 from Bothrops jararacussu

“…Figure 3 displays that erythrocytes treated with ADNT are highly susceptible to the hemolytic action of BthTx-II and this is since snake venoms and their fractions such as PLA2 can cause hemolysis by increasing cellular oxidative stress [56,57]. In addition, ADNT activity on these already well-known antioxidants thiol-dependent system and their inhibition effects reveal their crucial participation in the erythrocytes protection against the effects of oxidative stress by these proteins [58,43,59,14].…”