Transcriptome Analysis of Kidneys in a Mouse Model of Sepsis-Induced Acute Kidney Injury
This study delves into the pathophysiological underpinnings of sepsis-associated acute kidney injury (SA-AKI), a critical and often fatal complication arising from the systemic inflammatory response and immune dysregulation triggered by sepsis. By employing the cecal ligation and puncture (CLP) method to simulate sepsis-induced AKI in mice, we conducted comprehensive transcriptome profiling using RNA sequencing (RNA-seq) to explore the molecular dynamics and identify potential new mechanisms underlying SA-AKI. The comparative analysis between the CLP-induced AKI model and control subjects aims to unravel the complex interplay of genes and signaling pathways involved in kidney damage and failure during sepsis. Through this approach, our research seeks to contribute to the broader understanding of SA-AKI's molecular basis. Overall design: In this experiment, a total of six C57BL/6 mice were equally divided into two groups: a Cecal Ligation and Puncture (CLP) group and a Sham (control) group, each containing three mice. The objective was to investigate the transcriptomic changes in the kidneys of mice subjected to sepsis-induced acute kidney injury (AKI) compared to a control condition. For the induction of sepsis, the mice in the CLP group underwent a surgical procedure under aseptic conditions, where they were first anesthetized with 2% to 4% isoflurane. A midline incision of approximately 2 cm was made below the diaphragm to expose the cecum, which was then partially ligated (two-thirds) using a 5-0 silk suture and punctured twice with a 22-gauge needle to simulate sepsis. A gentle squeeze was applied to the cecum to ensure a small amount of feces was expelled through the puncture sites, effectively mimicking a sepsis condition. Following the procedure, the mice received 1 mL of saline subcutaneously for resuscitation. The Sham group underwent identical surgical procedures, with the exception of the cecal ligation and puncture, to serve as a control for surgical stress without inducing sepsis. This design allowed for the isolation of transcriptomic changes attributable to sepsis-induced AKI from those due to surgical intervention alone. Twenty-four hours post-operation, all mice were euthanized, and their kidneys were harvested for RNA sequencing analysis.