Simvastatin and Benznidazole-Mediated Prevention of Trypanosoma cruzi-Induced Endothelial Activation: Role of 15-epi-lipoxin A4 in the Action of Simvastatin

Abstract

Trypanosoma cruzi is the causal agent of Chagas Disease that is endemic in Latin American, afflicting more than ten million people approximately. This disease has two phases, acute and chronic. The acute phase is often asymptomatic, but with time it progresses to the chronic phase, affecting the heart and gastrointestinal tract and can be lethal. Chronic Chagas cardiomyopathy involves an inflammatory vasculopathy. Endothelial activation during Chagas disease entails the expression of cell adhesion molecules such as E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) through a mechanism involving NF-κB activation. Currently, specific trypanocidal therapy remains on benznidazole, although new triazole derivatives are promising. A novel strategy is proposed that aims at some pathophysiological processes to facilitate current antiparasitic therapy, decreasing treatment length or doses and slowing disease progress. Simvastatin has anti-inflammatory actions, including improvement of endothelial function, by inducing a novel pro-resolving lipid, the 5-lypoxygenase derivative 15-epi-lipoxin A4 (15-epi-LXA4), which belongs to aspirin-triggered lipoxins. Herein, we propose modifying endothelial activation with simvastatin or benznidazole and evaluate the pathways involved, including induction of 15-epi-LXA4. The effect of 5 μM simvastatin or 20 μM benznidazole upon endothelial activation was assessed in EA.hy926 or HUVEC cells, by E-selectin, ICAM-1 and VCAM-1 expression. 15-epi-LXA4 production and the relationship of both drugs with the NFκB pathway, as measured by IKK-IKB phosphorylation and nuclear migration of p65 protein was also assayed. Both drugs were administered to cell cultures 16 hours before the infection with T. cruzi parasites. Indeed, 5 μM simvastatin as well as 20 μM benznidazole prevented the increase in E-selectin, ICAM-1 and VCAM-1 expression in T. cruzi-infected endothelial cells by decreasing the NF-κB pathway. In conclusion, Simvastatin and benznidazole prevent endothelial activation induced by T. cruzi infection, and the effect of simvastatin is mediated by the inhibition of the NFκB pathway by inducing 15-epi-LXA4 production.

Simvastatin Attenuates Endothelial Activation through 15-Epi-Lipoxin A4 Production in Murine Chronic Chagas Cardiomyopathy

“…Recently, our group reported that aspirin decreases EC activation and myocardial inflammation in a T. cruzi-infected murine model via the production of 15-epi-LXA4 (16). In addition, simvastatin and benznidazole prevent EC activation in human umbilical vein endothelial cells (HUVECs), decreasing ECAM expression on the cell surface, an effect mediated by 15-epi-LXA4 (17). However, the role of simvastatin in endothelial activation and its link with 15-epi-LXA4 in vivo remain to be studied.…”

Notch receptor expression in Trypanosoma cruzi‐infected human umbilical vein endothelial cells treated with benznidazole or simvastatin revealed by microarray analysis

“…These manifestations are due to a chronic inflammatory process (Rassi et al, 2017). As part of these inflammatory processes, endothelial activation is observed, along with high expression of endothelial adhesion molecules that facilitate the recruitment of leukocytes that infiltrate the cardiac muscle (Campos‐Estrada et al, 2015). In addition, there is an increase in the production of thromboxane and platelet aggregation and the generation of local microthrombi and focal areas of ischemia, which may contribute to the appearance of fibrous tissue by myofibroblasts.…”

“…Previously, our group reported that acetylsalicylic acid (ASA) and simvastatin (Simv) modulate the expression of adhesion molecules in endothelial cells infected with T. cruzi (Molina‐Berrios et al, 2013a, b; Campos‐Estrada et al, 2015). Through this effect, the recruitment of inflammatory cells in infected tissues can be reduced.…”

“…In that model, inflammatory cell infiltration decreased when the mice were treated with these two drugs. However, it is not clear whether ASA or Simv, alone or in combination with Bz, affects the expression of endothelial adhesion molecules by modulating gene expression or by post‐transcriptional effects (Campos‐Estrada et al, 2015). Moreover, treating human umbilical vein endothelial cells (HUVECs) with anti‐inflammatory or trypanocidal drugs before the T. cruzi challenge allows us to determine, to some extent, the conditions during parasite dissemination throughout treatment; such conditions might occur after a late diagnosis of the disease (Campos‐Estrada et al, 2015).…”

“…However, it is not clear whether ASA or Simv, alone or in combination with Bz, affects the expression of endothelial adhesion molecules by modulating gene expression or by post‐transcriptional effects (Campos‐Estrada et al, 2015). Moreover, treating human umbilical vein endothelial cells (HUVECs) with anti‐inflammatory or trypanocidal drugs before the T. cruzi challenge allows us to determine, to some extent, the conditions during parasite dissemination throughout treatment; such conditions might occur after a late diagnosis of the disease (Campos‐Estrada et al, 2015). Throughout the infective process, during the acute and chronic phases, the cells are infected in a nonuniform manner; however, there is an inconstant but permanent spread of the parasite, from infected to uninfected cells, which may be near or far.…”