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Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction
Published on: August 27, 2020
Screening and identification of the differentially expressed proteins in neonatal rat kidney after partial unilateral
Qi Zhao1, Yansheng Xue2, Yi Yang1
1Department of Pediatric Surgery, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
Abstract:
Renal fibrosis, considered to be a common consequence of progressive renal disease, involves glomerulosclerosis and/or tubulointerstitial fibrosis. Currently, research is focused on investigating potential mechanisms to prevent or reverse the damage caused by fibrosis. Under the influence of cytokines, chemokines and other signaling molecules, the cellular interactions that regulate the development of interstitial fibrosis are complex. Epithelial‑mesenchymal transition (EMT) has emerged as an important pathway leading to the generation of matrix‑producing fibroblasts and myofibroblasts in diseased kidneys. The proteomics study compared the protein profiles between the time points of podocyte EMT and tubular cell EMT in a partial unilateral ureteral obstruction (PUUO) model in rats. Proteins isolated from the PUUO group and corresponding sham rat kidney tissues were subjected to 2‑D gel electrophoresis and were then identified by mass spectrometry. In total, 43 proteins with differential expression were identified, which were reported to be involved in the regulation of the cytoskeleton and actin, glucose metabolism, cell apoptosis, mitochondrial energy metabolism, oxidative stress and endoplasmic reticulum stress. Electron transfer flavoprotein, β polypeptide was detected by immunoblot analysis and its mRNA levels were determined in renal tissues. The results demonstrate protein alterations that reflect the pathology of the obstructed kidneys, and thus may aid in understanding the pathogenesis of obstructive nephropathy.
Insights
This study used proteomics to identify 43 differentially expressed proteins in rat kidneys with obstructive nephropathy. These proteins offer insights into the mechanisms of renal fibrosis and potential therapeutic targets.
Area of Science:
- Nephrology
- Proteomics
- Molecular Biology
Background:
- Renal fibrosis, a common outcome of kidney disease, involves glomerulosclerosis and tubulointerstitial fibrosis.
- Epithelial-mesenchymal transition (EMT) is a key pathway generating matrix-producing cells in kidney fibrosis.
- Understanding the molecular mechanisms of renal fibrosis is crucial for developing treatments.
Purpose of the Study:
- To compare protein profiles during podocyte and tubular cell EMT in a rat model of partial unilateral ureteral obstruction (PUUO).
- To identify proteins involved in the pathogenesis of obstructive nephropathy.
Main Methods:
- Proteomics analysis using 2-D gel electrophoresis and mass spectrometry on kidney tissues from PUUO and sham rats.
- Identification of differentially expressed proteins.
- Immunoblot analysis and mRNA level determination for specific proteins.
Main Results:
- Identified 43 differentially expressed proteins in obstructed kidneys compared to controls.
- These proteins are involved in cytoskeleton regulation, glucose metabolism, apoptosis, mitochondrial function, and stress responses.
- Electron transfer flavoprotein, β polypeptide was specifically analyzed.
Conclusions:
- Protein alterations in obstructed kidneys reflect the disease pathology.
- The identified proteins may contribute to understanding obstructive nephropathy pathogenesis.
- This proteomic data provides potential targets for preventing or reversing renal fibrosis.

