Abstract: PO0421
Proteogenomic Effects of Environmental and Uremic Toxin Acrolein on Mouse Kidneys
Session Information
- AKI: Repair and Progression
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Rane, Sanjana, University of Louisville, Louisville, Kentucky, United States
- Jin, Shunying, University of Louisville, Louisville, Kentucky, United States
- Isaacs, Susan M., University of Louisville, Louisville, Kentucky, United States
- Barati, Michelle T., University of Louisville, Louisville, Kentucky, United States
- Merchant, Michael, University of Louisville, Louisville, Kentucky, United States
- Pan, Jianmin, University of Louisville, Louisville, Kentucky, United States
- Rai, Shesh, University of Louisville, Louisville, Kentucky, United States
- Srivastava, Sanjay, University of Louisville, Louisville, Kentucky, United States
- Rane, Madhavi J., University of Louisville, Louisville, Kentucky, United States
Background
Acrolein is present in the environment, water, food and is a uremic toxin endogenously produced through lipid peroxidation and polyamine oxidation. Its mechanism of action involves cellular thiol reactivity and glutathione depletion-induced oxidative stress. However, direct effects of acrolein on kidneys are not known. The current study conducted RNA-seq and proteomic analysis on mouse kidneys exposed to acrolein.
Methods
C57BL/6 mice were subjected to filtered-air (control) or inhaled-acrolein (1.0 ppm), 6h/day, 5 days/week for 12 weeks. Total-RNA and kidney homogenates from all mice were subjected to RNA-seq and proteomic analysis. Immunoblotting, H&E and Sirius-Red staining of kidneys was performed.
Results
RNA-seq analysis detected activation of profibrotic pathways including TGFb/TGFbR/SMAD3/TEAD4/Autotaxin and down-regulation of anti-fibrotic pathways including STAT5A/FGF21/PPARb/IL-22. Proteomic analysis identified fibrotic pathways, activation of oxidative stress, mitochondrial dysfunction, proteasomal degradation, apoptosis, and inflammatory pathways. Proteomic analysis demonstrated acrolein-decreased expression of antioxidant proteins, non-erythroid hemoglobin and glutathione synthetase (GSS), targets of transcription factor Nuclear Factor-Erythroid derived-2 (NF-E2). Accordingly, acrolein decreased renal NF-E2 expression and increased cleaved caspase-3 and profibrotic connective tissue growth factor (CTGF) expression. H&E staining demonstrated damaged tubules while Sirius-Red staining demonstrated increased collagen deposition in kidneys of acrolein exposed mice. Over-expression of NF-E2 in immortalized human renal proximal tubule (HK-11) cells, inhibited acrolein-induced CTGF expression and caspase-3 cleavage.
Conclusion
Our studies demonstrate activation of oxidative stress, apoptotic and profibrotic pathways in acrolein treated kidneys. Chronic exposures of acrolein may lead to progressive fibrosis-induced End Stage Renal Disease (ESRD). In vivo modulation of NF-E2 expression may slow-down progressive fibrosis-induced ESRD.
Funding
- Other NIH Support