NEAT1 Regulates Oxidative Stress Induced By Calcium Oxalate Crystals In Renal Tubular Epithelial Cells Via IRF1/TLR4 And Promotes Crystal Deposition

Aims: During the process of urinary stone formation,including crystal nucleation,crystal aggregation,crystal retention,and crystal growth,and the adhesion of calcium crystals in the tubular lumen can induce tubular epithelial damage;this tubular epithelial damage can promote processes such as crystal adhesion and aggregation,accelerating stone formation.We found increased expression of long-stranded non-coding RNA NEAT1 in renal tubular epithelium with calcium oxalate deposition,along with simultaneous upregulation of inflammatory regulators of IRF1 and TLR4/NF-κB-related pathways.As a star molecule of inflammatory regulation,we hypothesized that NEAT1 is involved in calcium oxalate crystal-induced renal tubular epithelial cell injury,but the role of NEAT1 in this crystal deposition-related The regulation of NEAT1 in this crystal deposition-related injury has not been reported so far,and we focused on the regulatory mechanism of NEAT1.Methods: We used calcium oxalate crystals co-cultured with renal tubular epithelial cells HK-2 to create a cellular model,and animal models were created by administering glyoxalate to C57 male mice.Alterations in cellular injury and oxidative stress markers were observed by Western-Blot,RT-PCR,and cellular fluorescence staining.The damage induced by Ca Ox crystal deposition in cells and animal models was observed by tissue staining and scanning electron microscopy.Lastly,the targeting relationship and binding sites of NEAT1-mi R130-IRF1 were observed by in situ hybridization with nucleic acid probes and dual luciferase reporter gene assay.We inferred the regulatory relationship between NEAT1 and IRF1/TLR4/NF-κB,and explored potential drug targets and their effects in animal models in vivo experiments.Results: In the renal tissues of mice with renal calcium deposition injury model,NEAT1 expression was significantly elevated and positively correlated with IRF1,TLR4 and NF-κB.NEAT1 expression was always negatively correlated with mi R-130,and NEAT1 could inhibit its effect by adsorbing mi R-130.mi R-130 could bind to IRF1 3’-UTR and inhibit its translation process.When we inhibited the expression of NEAT1,IRF1,TLR4 and NF-κB were also inhibited to different degrees,while oxidative damage to renal tubular epithelial cells was also reduced.When we inhibited both NEAT1 and mi R-130 expression,the damage in mouse kidney tissue was again aggravated.Conclusion: In renal tubular injury induced by calcium oxalate crystal deposition,NEAT1 could release IRF1 through adsorption of mi RNA-130a-3p,and promote oxidative tubular epithelial damage during renal calcium deposition injury,and accelerate Ca Ox crystal deposition through TLR4/NF-κB and other pathways.NEAT1 inhibitors exhibit some pharmacotherapeutic effects,inhibiting TLR4 and other pathways that alleviate oxidative stress,but the effect on crystal deposition remains limited.