Fed State Prior to Hemorrhagic Shock and Polytrauma in a Porcine Model Results in Altered Liver Transcriptomic Response

<div><p>Hemorrhagic shock is a leading cause of trauma-related mortality in both civilian and military settings. Resuscitation often results in reperfusion injury and survivors are susceptible to developing multiple organ failure (MOF). The impact of fed state on the overall response to shock and resuscitation has been explored in some murine models but few clinically relevant large animal models. We have previously used metabolomics to establish that the fed state results in a different metabolic response in the porcine liver following hemorrhagic shock and resuscitation. In this study, we used our clinically relevant model of hemorrhagic shock and polytrauma and the Illumina HiSeq platform to determine if the liver transcriptomic response is also altered with respect to fed state. Functional analysis of the response to shock and resuscitation confirmed several typical responses including carbohydrate metabolism, cytokine inflammation, decreased cholesterol synthesis, and apoptosis. Our findings also suggest that the fasting state, relative to a carbohydrate prefed state, displays decreased carbohydrate metabolism, increased cytoskeleton reorganization and decreased inflammation in response to hemorrhagic shock and reperfusion. Evidence suggests that this is a consequence of a shrunken, catabolic state of the liver cells which provides an anti-inflammatory condition that partially mitigates hepatocellar damage.</p></div

Graphical representation of the experimental timeline described in Methods.

<p>Graphical representation of the experimental timeline described in Methods.</p

Heatmap of log2 fold changes of genes associated with peptidase activity in carbohydrate prefed (CPF) animals at each resuscitation timepoint (2, 8, and 20 hours) relative to Baseline (B).

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes increased mRNA expression with respect to Baseline whereas red denotes the opposite.</p

Heatmap of log2 fold changes of genes associated with cytoskeleton processes between carbohydrate prefed (CPF) and fasted (FS) animals.

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes higher concentration or larger changes in mRNA concentration in CPF whereas red denotes the opposite.</p

Diagram of simplified Interleukin-10 (IL-10) pathway.

<p>A hyperosmotic condition induces cellular shrinkage and activates tyrosine phosphorylation of Janus kinases (JAK) to subsequently phosphorylate STAT3 and IL-10 production.</p

Heatmap of log2 fold changes of genes associated with cytokine related genes in carbohydrate prefed (CPF) animals at each resuscitation timepoint (2, 8, and 20 hours) relative to Baseline (B).

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes increased mRNA expression with respect to Baseline whereas red denotes the opposite.</p

Heatmap of log2 fold changes of genes associated with cytoskeleton processes in fasted (FS) animals at each resuscitation timepoint (2, 8, and 20 hours) relative to Baseline (B).

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes increased mRNA expression with respect to Baseline whereas red denotes the opposite.</p

Number of differentially expressed genes between CPF and FS animals and between timepoints within groups.

<p>Number of differentially expressed genes between CPF and FS animals and between timepoints within groups.</p

Heatmap of log2 fold changes of genes associated with carbohydrate metabolism in fasted (FS) animals at each resuscitation timepoint (2, 8, and 20 hours) relative to Baseline (B).

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes increased mRNA expression with respect to Baseline whereas red denotes the opposite.</p

Heatmap of log2 fold changes of genes associated with ion transport in fasted (FS) animals at each resuscitation timepoint (2, 8, and 20 hours) relative to Baseline (B).

<p>Rows are differentially expressed genes (DEGs) following RNA sequencing. Columns denote the respective timepoints Baseline (B), 2 hours full resuscitation change from Baseline (FR2vB), 8 hours full resuscitation change from Baseline (FR8vB), and 20 hours full resuscitation change from Baseline (FR20vB). Green denotes increased mRNA expression with respect to Baseline whereas red denotes the opposite.</p