Conclusion
This study confirmed that NAT10 inhibits RTECs oxidative stress and cell pyrodeath through the enhanced ac4C modification of ULK1 and impedes kidney stone progression.
Material and methods
A kidney-stone cell model was simulated using calcium oxalate monohydrate (COM) in vitro. Western blot analysis of NAT10 expression and N4-acetylcytidine RNA immunoprecipitation verified the regulatory efficacy of NAT10 in Unc-51 like autophagy activating kinase 1 (ULK1) ac4C modification. The luciferase reporter gene assay further verified the interaction between NAT10 and ULK1. A kidney stone model was established using BALB/c mice injected with glyoxylic acid.
Methods
A kidney-stone cell model was simulated using calcium oxalate monohydrate (COM) in vitro. Western blot analysis of NAT10 expression and N4-acetylcytidine RNA immunoprecipitation verified the regulatory efficacy of NAT10 in Unc-51 like autophagy activating kinase 1 (ULK1) ac4C modification. The luciferase reporter gene assay further verified the interaction between NAT10 and ULK1. A kidney stone model was established using BALB/c mice injected with glyoxylic acid.
Objective
Among the most common chronic kidney diseases, kidney stones are second only to hypertension. Kidney stones pose a public health threat due to their increased incidence, high recurrence rate, and heavy economic burden. In this work, we investigated the potential mechanism of N-acetyltransferase 10 (NAT10) in oxidative stress and pyroptosis of renal tubular epithelial cells (RTECs). Material and
Results
COM can dose-dependently suppressed the cell viability and superoxide dismutase activity of HK-2 cells and promoted the release of lactate dehydrogenase and malondialdehyde levels (P < 0.05). COM also promoted apoptosis in HK-2 cells, upregulated the protein levels of caspase-1 and gasdermin D-N, and simultaneously enhanced the HK-2 cell secretion of interleukin-1b (IL-1b) and IL-18 (P < 0.05). The overexpression of NAT10 in HK-2 cells reversed the aforementioned effects, and that of NAT10 upregulated the messenger RNA (mRNA) levels of ULK1 and increased ac4C modification (P < 0.01). Furthermore, only the luciferase activity of the wild-type ULK1 containing NAT10 binding sites was enhanced with the upregulation of NAT10 (P < 0.001). Actinomycin D treatment showed that NAT10 overexpression extended the half-life of ULK1 mRNA (P < 0.01). Silencing of ULK1 neutralized the effects of NAT10 overexpression on COM-induced cell injury (P < 0.05). In addition, the increased expression of NAT10 inhibited crystal deposition, oxidative stress, and apoptosis in vivo (P < 0.05).