Abstract: PO0146
Microparticles Released in Response to AKI May Influence Glucose and Salt Metabolism
Session Information
- AKI Mechanisms - 1
October 22, 2020 | Location: On-Demand
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Campos, Begoña, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
- Harrison, Kathleen, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
- Kramer, Samantha M., University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
- Abu Jawdeh, Bassam G., University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
- Thakar, Charuhas V., University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
Background
Renal epithelial injury due to ischemia or toxic/inflammatory insults is the primary cause of acute kidney injury (AKI). Due to their high metabolic activity renal tubular epithelial cells (RPTEC) are especially vulnerable to damage leading to a clinical syndrome of disruption in salt, water and glucose homeostasis. We have previously shown that microparticles (MP) derived from renal epithelial cells are released in the setting of kidney injury and can be detected in vitro as well as in human plasma, and can carry the biological activity.
Methods
In this study, we evaluated the release of MP carrying SGLT2, and SLC12A3 (NCC) in response to kidney injury. Immortalized RPTEC lines were treated with Oxidative stress (H2O2) or inflammatory stress (TNFα) agents by the validated methodology to study in vitro models of AKI. Human samples were derived from a prospectively collected repository (31 cases of AKI in critically ill patients compared to 22 living kidney donor healthy controls). Samples were prepared to measure MP (standard methods), and flow cytometric analysis was evaluated using antibodies against SGLT2, and SLC12A3 (NCC). FlowJo software was used for analysis. Mann-Whitney test was used for comparisons.
Results
RPTEC models of injury (H2O2) resulted in a significant increase in the release of MP positive for SGLT2 MP 5.49 X 1010/ml vs 0.41 X 1010/ml for control cells (p=0.05) and NCC (SLC12A3) 4.2 x 105 vs 0.63 X 105 for control cells (p=0.05). Similar changes were observed when cells were treated with TNFα (p=0.05) for SGLT2 and NCC.
We also confirmed the presence of MP containing SGLT2 and NCC in AKI (104.60 X105 /ml and 1.77 X 105 /ml, respectively). However, when compared to controls, the difference was statistically similar.
Conclusion
This is one of the first reports to confirm that key transporters from renal epithelium can be released and detected as MP in both in vitro and clinical settings of AKI. These findings may provide novel insights into the mechanisms of glucose, salt and water dysregulation during kidney injury and repair.