Abstract: PO0710
Downregulation of Ehhadh and Tubular Dysfunction in Diabetic Nephropathy
Session Information
- Diabetic Kidney Disease: Basic - II
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Diabetic Kidney Disease
- 601 Diabetic Kidney Disease: Basic
Authors
- Zhong, Jianyong, Vanderbilt University Medical Center, Nashville, Tennessee, United States
- Yang, Haichun, Vanderbilt University Medical Center, Nashville, Tennessee, United States
- Harris, Raymond C., Vanderbilt University Medical Center, Nashville, Tennessee, United States
- Fogo, Agnes B., Vanderbilt University Medical Center, Nashville, Tennessee, United States
Background
As a peroxisomal protein, enoyl-CoA hydratase and 3-hydroxyacyl CoA dehydrogenase (Ehhadh) catalyzes the second and third committed steps in the peroxisomal beta-oxidation pathway. Ehhadh also interacts with catalase and peroxisomal biogenesis factor 5 (PEX5), which decomposes hydrogen peroxide and regulates peroxisomal biogenesis, respectively. Previously we detected reduced tubular Ehhadh expression in human and mouse diabetic nephropathy. This study aims to investigate the potential impact of Ehhadh on peroxisomal/mitochondria functional change in response to high glucose in vitro.
Methods
Primary cultured proximal tubular epithelial cells (PTC) were exposed to high glucose, mannitol or control medium. Ehhadh subcellular localization and peroxisome quantitation (area per cell) were analyzed by confocal microscopy. Ehhadh, catalase, PEX5, ACOX1, Hsd17d4, scp2, ACAA1, cpt1a, Acadm, AHDHB, Acat1 and ACAA2 mRNA expression were assessed by qPCR. Peroxidase activity and oxidase stress were also analyzed.
Results
Ehhadh transcription and protein were significantly downregulated in PTC under high glucose conditions. Ehhadh was localized mostly to peroxisomes and rarely in mitochondria. Key enzymes for beta-oxidation in peroxisomes (ACOX1, Hsd17d4, scp2 and ACAA1) and mitochondria (cpt1a, Acadm, AHDHB, Acat1 and ACAA2) were not changed under high glucose conditions. Catalase transcription and peroxidase activity were reduced in high glucose vs control. PEX5 was also reduced, but peroxisome quantitation was increased 39.6% under high glucose conditions. Oxidative stress was also increased 7.6% in high glucose vs control.
Conclusion
Ehhadh downregulation is associated with reduced peroxidase activity, increased peroxisomal biogenesis and oxidative stress in PTC. Whether altering Ehhadh can impact such dysfunction awaits further study.
Funding
- NIDDK Support