Preliminary profiling of blood transcriptome in a rat model of hemorrhagic shock

Abstract

Hemorrhagic shock is a leading cause of morbidity and mortality worldwide. Significant blood loss may lead to decreased blood pressure and inadequate tissue perfusion with resultant organ failure and death, even after replacement of lost blood volume. One reason for this high acuity is that the fundamental mechanisms of shock are poorly understood. Proteomic and metabolomic approaches have been used to investigate the molecular events occurring in hemorrhagic shock but, to our knowledge, a systematic analysis of the transcriptomic profile is missing. Therefore, a pilot analysis using paired-end RNA sequencing was used to identify changes that occur in the blood transcriptome of rats subjected to hemorrhagic shock after blood reinfusion. Hemorrhagic shock was induced using a Wigger's shock model. The transcriptome of whole blood from shocked animals shows modulation of genes related to inflammation and immune response (Tlr13, Il1b, Ccl6, Lgals3), antioxidant functions (Mt2A, Mt1), tissue injury and repair pathways (Gpnmb, Trim72) and lipid mediators (Alox5ap, Ltb4r, Ptger2) compared with control animals. These findings are congruent with results obtained in hemorrhagic shock analysis by other authors using metabolomics and proteomics. The analysis of blood transcriptome may be a valuable tool to understand the biological changes occurring in hemorrhagic shock and a promising approach for the identification of novel biomarkers and therapeutic targets. Impact statement This study provides the first pilot analysis of the changes occurring in transcriptome expression of whole blood in hemorrhagic shock (HS) rats. We showed that the analysis of blood transcriptome is a useful approach to investigate pathways and functional alterations in this disease condition. This pilot study encourages the possible application of transcriptome analysis in the clinical setting, for the molecular profiling of whole blood in HS patients.

Transcriptome Characterization of Short Distance Transport Stress in Beef Cattle Blood

“…To ensure the quality of RNA in samples, the following steps were performed as quickly as possible. ACK lysis buffer lysed erythrocytes, karyocytes were harvested following centrifugation at 4,000 rpm for 6 min at 4°C, and then transfer into eppendorf tubes containing trizol, and finally stored in liquid nitrogen ( Braga et al, 2017 ; Huang et al, 2017 ). Total RNA from the blood samples ( n = 7) were isolated from leukocytes using the RNAiso Plus Kit (Takara, Tokyo, Japan).…”

Human cell-camouflaged nanomagnetic scavengers restore immune homeostasis in a rodent model with bacteremia

“…We also tested whether the acquisition of antibiotic resistance in gram-positive ( S. aureus , MRSA, and vancomycin-intermediate Staphylococcus aureus (VISA)) and gram-negative ( E. coli , ESBL-positive E. coli , and carbapenem-resistant (CR) E. coli ) bacteria compromises the capturing efficiency of MNVs due to alteration of the recognized molecules present on the bacterial cell wall. Despite the reported modification of PBP2a on MRSA and cell wall peptidoglycan on VISA, [65,66] which was induced by the acquisition of antibiotic resistance, the capturing efficiency of hRBC-MNVs to all three strains was retained (Figure 3c), which could be attributed to the interaction between the glycophorins and the mucin-binding proteins in S. aureus . [67-69] Similar binding characteristics were also observed for the gram-negative bacteria in which E. coli , ESBL-positive E. coli , and CR E. coli were depleted in the blood at the same rate (Figure 3d).…”

“…Notably, 83 genes among the 420 significantly downregulated genes were associated with sepsis, such as S100a8, -9, [84] Lcn2, [85] Pf4, [86] Gzma, Gzmb, [87] and Olfm4, [88] which are known to be upregulated in septic or severe trauma patients; in contrast, only one gene out of the 10 significantly upregulated genes was identified as sepsis-associated by the Open Targets Platform, which was RT1-Da. [89] Similar to RT1-Da, several other upregulated genes were found to be downregulated in septic patients, including CD74, [90] Rps27, Rpl17, and Rpl3 (Figure 5i). [91,92] Moreover, for the top 10 terms of gene ontology analysis in the downregulated biological process, differentially expressed genes with the highest significance in comparison to those in the untreated animals ( P < 0.001) were associated with the response to external stimulus, response to stress, cell death and biological process involved in interspecies between organisms, supporting the potential clinical impact of our blood-cleansing system, which not only improves mortality but also reinstates the homeostasis of immunity (Figure 5j).…”

Human Cell‐Camouflaged Nanomagnetic Scavengers Restore Immune Homeostasis in a Rodent Model with Bacteremia

“…Notably, 83 genes among the 420 significantly downregulated genes were associated with sepsis, such as S100a8, ‐9, [ 78 ] Lcn2, [ 79 ] Pf4, [ 80 ] Gzma, Gzmb, [ 81 ] and Olfm4, [ 82 ] which are known to be upregulated in septic or severe trauma patients; in contrast, only one gene out of the 10 significantly upregulated genes was identified as sepsis‐associated by the Open Targets Platform, which was RT1‐Da. [ 83 ] Similar to RT1‐Da, several other upregulated genes were found to be downregulated in septic patients, including CD74, [ 84 ] Rps27, Rpl17, and Rpl3 (Figure 5i). [ 85,86 ] Moreover, for the top 10 terms of gene ontology analysis in the downregulated biological process, differentially expressed genes with the highest significance in comparison to those in the untreated animals ( P < 0.001) were associated with the response to external stimulus, response to stress, cell death and biological process involved in interspecies between organisms, supporting the potential clinical impact of our blood‐cleansing system, which not only improves mortality but also reinstates the homeostasis of immunity (Figure 5j).…”