Profiling Circulating MicroRNA Expression in Experimental Sepsis Using Cecal Ligation and Puncture

Abstract

The levels of circulating microRNAs (miRNAs) in mice with experimental sepsis induced by cecal ligation and puncture (CLP) were determined using whole blood samples obtained from C57BL/6 mice at 4, 8, and 24 h after CLP; miRNA expression analysis was performed in these samples using an miRNA array. Microarray analysis revealed upregulation of 10 miRNA targets (miR-16, miR-17, miR-20a, miR-20b, miR-26a, miR-26b, miR-106a, miR-106b, miR-195, and miR-451). The expression of these miRNA targets in the whole blood, serum, and white blood cells (WBCs) of CLP mice was quantified using quantitative real-time PCR; these values were compared to those in sham-operated C57BL/6 mice, and the results indicated that these miRNA targets were significantly up-regulated in the whole blood and serum but not in the WBCs. In addition, the levels of these 10 miRNA targets in the serum of Tlr2−/−, Tlr4−/−, and NF-κB−/− mice at 8 h after CLP did not decrease significantly., which indicated that the transcription of these miRNAs was not directly mediated by the TLR2/NF-κB or TLR4/NF-κB pathway, and pathways induced by exposure to the gram-positive or gram-negative bacteria. Immunoprecipitation with the Argonaute 2 ribonucleoprotein complex revealed significantly increased expression of the 10 miRNA targets in the serum of mice after CLP, and the levels of 6 (miR-16, miR-17, miR-20a, miR-20b, miR-26a, and miR-26b) of these 10 miRNA targets increased significantly in exosomes isolated using ExoQuick precipitation solution. In this study, we identified circulating miRNAs that were up-regulated after CLP and determined the increase in the levels of these miRNAs, and our results suggest that circulating Ago2 complexes and exosomes may be responsible for the stability of miRNAs in the serum.

Extracellular microRNA 130b‐3p inhibits eCIRP‐induced inflammation

“…These miRNAs are preserved in circulation and are quantifiable, which supports the potential role as diagnostic or progression biomarkers in various diseases [13,24,25]. These miRNAs are preserved in circulation and are quantifiable, which supports the potential role as diagnostic or progression biomarkers in various diseases [13,24,25].…”

Proteomics study of serum exosomes in Kawasaki disease patients with coronary artery aneurysms

“…Exosomes were separated from the sera of all participants using ExoQuick precipitation (System Biosciences Inc., Palo Alto, CA, USA) according to the manufacturer instructions [13,14].…”

Identification and validation of microRNAs as endogenous controls for quantitative polymerase chain reaction in plasma for stable coronary artery disease

“…miR-16 is also a universal reference gene because of its high expression level. But some recent studies reported up-regulation of miR-16 in plasma in some diseases, such as autoimmune thyroid disease [26], experimental sepsis [27] and gastric cancer [28]. Other research demonstrated that miR-16 is highly expressed in red blood cells so that any hemolysis of blood samples strongly affects their level in plasma [29].…”