Effect of nitro fatty acids in LPS challenged primary human renal tubular epithelial cells

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Introduction
Nitro-oleic acids are endogenous lipid products with potent anti-inflammatory properties mediating diverse cell signaling and pathologic processes in amongst others renal diseases. Previous studies suggested that NFAs exert a renoprotective effect during inflammatory processes also in acute kidney injury, which occurs pathophysiologically mainly within the renal tubular system. Therefore, we investigated the regulatory effect of NFAs on primary human renal tubular epithelial cells (TEC) in vitro based on an acute damage model with LPS.
Material and Methods
TECs were isolated from healthy renal tissue from four patients undergoing tumor nephrectomies and used in passage 1-4. After 3h starving, cells were pretreated for 30 min with either oleic acid (OA) as control substance or 9-nitro oleic acid (9-NO) at 1µM and 5µM. Hereafter TECs were exposed to LPS (1µg/ml) and harvested subsequent to 3h of incubation.
Results
We first evaluated adequate dosing of lipopolysaccharides (LPS) and 9-NO on TECs, which provokes damage but also allows a subsequent cellular regeneration. In the XTT assay we observed a significant reduced TEC viability along with increased LDH in the supernatant at higher 9-NO concentrations and concurrent LPS. This was confirmed by reduced impedance measurement using iCelligence and obvious rarefication of TECs in immunofluorescence Hoechst and Phalloidin staining. Regarding NFA specific signaling, Hemoxygenase-1 (HO-1), Cyclooxygenase-1 (COX-1), 5-lipoxygenase (5-LO) mRNA was regulated in TECs upon 9-NO and LPS treatment.
Conclusion
Our hitherto existing data indicate that NFAs and their signaling pathway are involved in acute renal tubular injury.
OriginalsprogEngelsk
Udgivelsesår2019
StatusUdgivet - 2019

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