Nesfatin‑1 Deficiency Exacerbates Calcium Oxalate Monohydrate-Induced Cellular Injury in Renal Epithelial Cells.

Abstract

Calcium oxalate monohydrate (COM) crystals are major contributors to nephrolithiasis-associated renal tubular injury, primarily through mechanisms involving oxidative stress, inflammation, apoptosis, and epithelial-mesenchymal transition (EMT). While nesfatin-1 is known for its antioxidant and cytoprotective actions, the specific role of endogenous nesfatin-1 in regulating COM-induced epithelial injury has not been fully defined. In this study, nesfatin-1 was silenced using antisense oligonucleotides in NRK-52E and primary renal tubular epithelial cells to examine how loss of endogenous nesfatin-1 peptide influences cellular susceptibility to COM. COM exposure reduced nesfatin-1 levels, and this decline was markedly greater in the nesfatin-1 silenced + COM group. Silencing nesfatin-1 intensified COM-driven oxidative stress, as reflected by reduced catalase activity and increased levels of malondialdehyde, nitric oxide, and ROS. Pro-inflammatory markers including NF-κB, TNF-α, IL-6, and IL-1β were upregulated, whereas IL-10 was decreased. Apoptotic indicators such as elevated intracellular Ca2+, diminished Bcl-2, increased caspase-3 activation, and higher proportions of apoptotic and necrotic cells were also evident. EMT and fibrotic markers (TGF-β1, α-SMA, Smad2/3, Smad4, vimentin) were increased, alongside reduced E-cadherin expression. Exogenous nesfatin-1 mitigated oxidative, inflammatory, apoptotic, and fibrotic responses in COM-treated cells; however, these protective effects were minimal in nesfatin-1 silenced + COM cells. Overall, the findings demonstrate that endogenous nesfatin-1 is indispensable for maintaining epithelial stability under COM-induced stress, and its absence amplifies cellular injury while diminishes responsiveness to exogenous peptide. Nesfatin-1 thus emerges as a potential therapeutic target for limiting crystal-induced renal cells damage.