Journal of Kidney

ISSN - 2472-1220


Cytotoxicity of Bothrops pauloensis Venom on Madin-Darby Canine Kidney Cells: Cell Death Mechanism

Marinho AD, Morais ICO, Lima DB, Pessoa Bezerra de Menezes RRP, Pereira GJS, Silveira JAM, Toyama MH, Orzáez M, Martins AMC and Monteiro HSA

Bothrops is a genus of pit vipers (snakes) endemic to Central and South America. Snake envenomation (Bothrops genus) specifically Bothropoides pauloensis, a venomous snake popularly known as “Jararaca pintada”, which is found in the Brazilian territory or more commonly in the southwest of the state of Sao Paulois. Acute kidney injury is one of the complications observed in Bothrops snakebite with relevant morbidity and mortality. Here, we showed the cytotoxic effect of Bothropoides pauloensis venom on MDCK cells, intracellular mitochondrial membrane potentiality (ΔΨm), BpV regulated Reactive Oxygen Species (ROS) and activation of caspases 3 and 7 in MDCK cells. MDCK cells were treated with different concentrations of B. pauloensis venom for 24 h and cell death were measured with annexin V and PI staining and detected by flow cytometry. MDCK cells were treated with two different concentrations (IC50 and 2 × IC50) of BpV after 24 h. All data are expressed as mean ± SEM of three independent experiments with six replicates (ANOVA and Dunnett test, *p<0.05). Cytotoxicity was assessed by MTT assay and the treatment with BpV caused decrease in cell viability, with an IC50 of 7.5 μg/mL. After the treatment, amounts of ROS in the BpV-treated MDCK-cells (7.5 μg/mL) showed significant right dislocation of the fluorescence peak in the histogram, when compared with the untreated control group. Caspase 3 and 7 activity was determined in presence of the fluorogenic Ac-DEVDafc substrate after treatment with Bothropoides pauloensis venom (12 h) then the result indicated apoptotic involvement in BpV-induced cell death. Similar results were also found for venoms of other Bothrops and Bothropoides genus snakes such as B. alternatus, B. insularis and B. leucurus.