Gene Microarray Integrated with High-Throughput Proteomics for the Discovery of Transthyretin in Rhabdomyolysis-Induced Acute Kidney Injury

Abstract

Background/Aims: Rhabdomyolysis, one of the leading causes of acute kidney injury (AKI), develops after trauma, drug toxicity, infections, burns, and physical exertion. The aim of this study was to investigate differences in gene and protein expression to elucidate the pathogenesis of rhabdomyolysis (RM)-induced AKI. Methods: In this study, we used glycerol induced renal injury as a model of RM-induced AKI. Affymetrix U133 plus 2.0 microarrays were used to perform gene microarray analysis. Isobaric tagging with related and absolute quantitation (iTRAQ) labeling mass spectrometry (MS) was applied to screen and identify differentially expressed proteins between RM-induced AKI and normal murine renal tissue. Verification experiments included immunohistochemistry (IHC), real-time PCR, Western blotting, and the measurement of ATP and ROS production. HK-2 cells were incubated in vitro with ferrous myoglobin and pcDNA-TTR, followed by assays to detect cell proliferation, ROS and apoptosis. Results: According to gene microarray and iTRAQ-MS analysis, we screened 17 common elements. After multiple analyses, we selected transthyretin (TTR) as our focus and investigated TTR in the kidney. Verification experiments with IHC confirmed differential expression levels of TTR proteins. Furthermore, Western blotting showed a stepwise decrease in TTR in AKI renal tissues. Cell-based experiments showed that overexpression of TTR could improve HK-2 cell viability and inhibit apoptosis. TTR reduced apoptosis by decreasing the accumulation of reactive oxygen species (ROS). Conclusion: This study reports a possible mechanism for RM-induced AKI and suggests that reductions in TTR could increase the generation of ROS and induce apoptosis. TTR may be a potentially valuable target for RM-induced AKI.

Mitophagy Plays a Protective Role in Iodinated Contrast-Induced Acute Renal Tubular Epithelial Cells Injury

“…Acute kidney injury (AKI) is a clinical syndrome with abnormal renal function or structure that occurs within 48 h and the leading clinical cause of chronic kidney disease, with high morbidity and mortality [1-3]. With the widely utilization of contrast media in the angiography and percutaneous coronary intervention, contrast-induced acute kidney injury (CI-AKI) has generally been considered as the third most common cause of hospital-acquired acute kidney injury (AKI) [4].…”

Sequential Organ Failure Assessment Score for Prediction of Mortality of Patients With Rhabdomyolysis Following Exertional Heatstroke: A Longitudinal Cohort Study in Southern China

“…In addition, some nuclear proteins are released by muscle cell injuries, such as histone 3 ( 19 , 20 ) and HMGB1 ( 21 ), which can activate platelets and then lead to the occurrence and development of DIC and ALI. Previous studies have found that the renal tubular injury in RM caused by crush syndrome is mainly caused by apoptosis ( 22 ), which shows that RM is prone to AKI. The mechanism of its occurrence may include the following two aspects or the result of a combination: the deficiency of effective circulating blood volume caused by fluid loss and dehydration ( 23 ), and the mechanical obstruction of renal tubules caused by MB released RM ( 24 ).…”

Exogenous biological renal support improves kidney function in rhabdomyolysis-induced acute kidney injury in mice

“…Tubular changes are mainly sublethal 24 hours after the onset of RM [28,29]. By 72 hours, the glyceroltreated mice developed AKI and exhibited signi cantly increased Scr and BUN levels and signi cant morphological changes [25].…”