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Updated: May 19, 2026
A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
Published on: September 11, 2020
MicroRNA expression following activated protein C treatment during septic shock
Cathy C Moore1, Iain H McKillop, Toan Huynh
1Division of Research, Department of Surgery, Carolinas Medical Center, Charlotte, NC 28232, USA.
Background:
Sepsis induces systemic stress by augmenting inflammatory and procoagulant responses, resulting in microvascular dysfunction and end organ failure, events modulated by the protein C pathway. MicroRNAs (miRNAs) are small noncoding RNAs involved in post-transcriptional regulation of genes; yet, their role in sepsis is poorly defined. We hypothesized that activated protein C (aPC) selectively alters specific miRNA expression implicated in protection of hepatic function during septic shock.
Methods:
Male Sprague-Dawley rats underwent sham or cecal ligation and puncture surgery; 24 h later, we randomized them to aPC (1 mg/kg) or vehicle (0.9% [w/v] saline) treatment via an indwelling venous catheter (12-h intervals for 24 h). We performed gene array and quantitative reverse transcriptase-polymerase chain reaction analysis on hepatic RNA to determine miRNA expression and determined predicted mRNA targets using a bioinformatics approach. We confirmed beneficial effects of aPC treatment in the cecal ligation and puncture model of sepsis by survival and blood chemistries, and histologically.
Results:
Of 351 rat miRNAs examined, 17 were highly expressed during sepsis and restored to basal levels after aPC treatment. We confirmed expression of select miRNAs (miR-182, -199a-5p, -203, -211, -222, and -29b) using quantitative reverse transcriptase-polymerase chain reaction. In silico analysis identified nine miRNAs significantly regulating target genes of the focal adhesion pathway.
Conclusions:
These data suggest that aPC treatment coordinates beneficial cytoprotective effects during sepsis by modulating miRNA expression. Whereas translational effects remain to be fully elucidated in a clinical setting, we demonstrate here the potential experimental and computational benefits of using of microRNA analysis in sepsis.
Insights
Activated protein C (aPC) treatment in sepsis modulates microRNA (miRNA) expression, offering protective effects on liver function. This study highlights miRNA analysis as a valuable tool for understanding sepsis mechanisms and potential therapies.
Area of Science:
- Molecular biology
- Genetics
- Biochemistry
Background:
- Sepsis causes systemic stress, inflammation, and organ failure, influenced by the protein C pathway.
- MicroRNAs (miRNAs) regulate gene expression, but their role in sepsis is not well understood.
- Activated protein C (aPC) may protect liver function during sepsis by altering miRNA expression.
Purpose of the Study:
- To investigate the hypothesis that aPC selectively alters specific miRNA expression in sepsis.
- To identify miRNAs involved in the protective effects of aPC on hepatic function during septic shock.
- To explore the potential of miRNA analysis in sepsis research.
Main Methods:
- Male Sprague-Dawley rats underwent cecal ligation and puncture (CLP) surgery to induce sepsis.
- Rats were treated with aPC or vehicle, and hepatic RNA was analyzed for miRNA expression using gene arrays and qRT-PCR.
- Bioinformatics was used to predict mRNA targets of identified miRNAs.
Main Results:
- Sepsis upregulated 17 out of 351 rat miRNAs, which were normalized by aPC treatment.
- Quantitative reverse transcriptase-polymerase chain reaction confirmed the expression of specific miRNAs (miR-182, -199a-5p, -203, -211, -222, -29b).
- In silico analysis revealed nine miRNAs regulating focal adhesion pathway genes.
Conclusions:
- aPC treatment demonstrates beneficial cytoprotective effects in sepsis by modulating miRNA expression.
- This study underscores the experimental and computational utility of miRNA analysis in sepsis.
- Further clinical research is needed to fully elucidate the translational effects of these findings.
