Use of dapagliflozin in Sepsis-induced Acute Kindey Injury

Abstract

Objective: to study the possible protection or not of dapagliflozin (SGLT2 inhibitor) in acute kidney injury induced by sepsis in non-diabetic animals. Glomerular and tubular functions will be assessed by creatinine clearance, ion excretion fractions, urinary osmolality, and urinary volume. Background: sepsis is a challenging, heterogeneous, and very varied clinical condition, being the main cause of death in intensive care units (ICU). The sepsis trigger is an infection that modifies the local and systemic functioning of immune-inflammatory reactions, resulting in multiple organ dysfunction and with high rates of mortality and morbidity. One of these dysfunctions, acute kidney injury (ARI), occurs in 50% of patients with sepsis, and the development of ARI consists of a poor prognosis for them, with consequent mortality in about 70%, doubling the mortality from sepsis. In sepsis-induced ARF, a series of evidence indicates that renal injury occurs without the presence of global or local hypoperfusion, but with increased renal blood flow and that it is mainly characterized by localized areas of slow peritubular blood flow and oxidative stress from cells tubular. In this context, it has been theorized that SGLT2 inhibitors (co-transporter of sodium and glucose in the renal proximal tubule) could reduce acute renal injury, in view of their renal protection effects already proven in both acute and chronic injuries, mainly by reducing glomerular filtration rate and oxidative stress reduction. Methods: the experimental procedures will be developed with the approval of the Ethics Committee on the Use of Animals (FMUSP - CAPPESQ) of the Faculdade de Medicina da Universidade de São Paulo, Brazil, in the use of animals. 24 male Wistar rats weighing between 200 - 250g and around 8 weeks of age will be used, which will be obtained from the Central Animal Hospital of FMUSP. The animals will be kept separate, in an environment with controlled temperature (22 - 24oC) and light (12 hours light / dark cycle) and will have free access to water and the AIN-93G standard diet. The animals will be randomly divided into three groups listed below. All treated animals will receive 5 mg/kg/day for gavage for 8 days and will be separated into three groups:3.1.1) Control: animals placed in a metabolic cage for 24 hours to collect blood, urine and remove the kidneys;3.1.2) Cecum ligation and perforation (CLP): they will receive 7 days of distilled water by gavage in the same amount as the dilution of dapagliflozin, on the 7th day there will be a surgical procedure for ligation and perforation of the cecum. After the procedure, they will be placed in a metabolic cage for 12h to collect blood, urine, and kidney removal;3.1.3) Cecum ligation and perforation being treated with dapagliflozin (CLP + DAPA) for 8 days: they will receive dapagliflozin for 7 days by gavage diluted in distilled water, on the 7th-day surgical procedure for ligation and perforation of the caecum and after that, they will be placed in a metabolic cage for 12 hours to collect blood, urine and remove the kidneys. The following parameters and procedures will be analyzed and performed: biochemical determinations (serum urea, serum/urinary sodium, dosage and fractions of cholesterol and serum / urinary creatinine), extraction of total proteins and renal tissue membrane (Western Blot), expression of proteins in kidney tissue, immunohistochemistry, electron microscopy in kidney tissue, quantitative and qualitative ultrastructural analysis, kidney injury score, the study of TUNEL cells in the kidney, histological evaluation of tubular injury in kidney tissue, mortality curve, and statistical analysis. (AU)