Evaluation of the therapeutic effect of methylene blue on the liver of rats submitted to ischemia and reperfusion.

Orlando Castro-E-Silva,Marina Rodrigues Garcia Silveira,Patricia Zorzi,Daniel Tófoli Queiroz Campos,Karina Dal Sasso Mendes,João Paulo Victorino,Maria Cecília Jordani,Luiz Augusto Carneiro D’Albuquerque,Paulo Roberto Barbosa Évora,Jordan Bistafa Liu

doi:10.1590/s0102-865020180120000001

Orlando Castro-E-Silva, Marina Rodrigues Garcia Silveira + Show 8 more

Open Access

https://doi.org/10.1590/s0102-865020180120000001

Copy DOI

Abstract

To analyze the effect of methylene blue (MB) therapy during the liver ischemia-reperfusion injury (I/R) process. Thirty-five male Wistar rats were used, (70%) submitted to partial ischemia (IR) or not (NIR) (30%) were obtained from the same animal. These animals were divided into six groups: 1) Sham (SH), 2) Sham with MB (SH-MB); 3) I/R, submitted to 60 minutes of partial ischemia and 15 minutes of reperfusion; 4) NI/R, without I/R obtained from the same animal of group I/R; 5) I/R-MB submitted to I/R and MB and 6) NI/R-MB, without I/R. Mitochondrial function was evaluated. Osmotic swelling of mitochondria as well as the determination of malondialdehyde (MDA) was evaluated. Serum (ALT/AST) dosages were also performed. MB was used at the concentration of 15mg/kg, 15 minutes before hepatic reperfusion. Statistical analysis was done by the Mann Whitney test at 5%. State 3 shows inhibition in all ischemic groups. State 4 was increased in all groups, except the I/R-MB and NI/R-MB groups. RCR showed a decrease in all I/R and NI/R groups. Mitochondrial osmotic swelling showed an increase in all I/R NI/R groups in the presence or absence of MB. About MDA, there was a decrease in SH values in the presence of MB and this decrease was maintained in the I/R group. AST levels were increased in all ischemic with or without MB. The methylene blue was not able to restore the mitochondrial parameters studied. Also, it was able to decrease lipid peroxidation, preventing the formation of reactive oxygen species.

Highlights

In several situations, the liver is submitted to periods of ischemia, as in hepatic trauma, transplantation, partial liver resections and circulatory shock
Thirty - five male Wistar rats weighing 200-300g were used, and in one mouse samples of blood and hepatic tissues were submitted to ischemia-reperfusion (I / R) and samples not submitted to ischemia-reperfusion (NI/R)
State 3 showed inhibition in all ischemic groups about the Sham group, which did not occur in the non-ischemic contralateral portion of the same liver, demonstrating inhibition of the flow of electrons in the respiratory chain

Summary

Introduction

The liver is submitted to periods of ischemia, as in hepatic trauma, transplantation, partial liver resections and circulatory shock. The ischemia and reperfusion injury (I/R) is related to local and systemic damage, which can result in loss or dysfunction of the organ, increased morbidity, and mortality of patients submitted to it. The damage caused by ischemia is related to the depletion of adenosine triphosphate (ATP), with activation of anaerobic metabolic pathways and the development of metabolic acidosis. Intracellular calcium accumulation, lysosomal rupture, and cell swelling occur. Reperfusion injury involves, in its early stage, mitochondrial dysfunction with the membrane permeability transition (TPM) of this organelle. Activation of the Kupffer cells occurs, with the release of more free radicals and pro-inflammatory cytokines. The lesion is marked by the activation of neutrophils[3,4,5,6]

Methods

Results

Conclusion