Jiang Hongda, Liu Yuting, Yu Yinggui, Yan Yu
Department of Laboratory Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, China.
Department of Ophthalmology, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, China.
J Biochem Mol Toxicol. 2025 Feb;39(2):e70150. doi: 10.1002/jbt.70150.
Age-related cataracts (ARCs) are associated with increased oxidative stress and cellular senescence. Our objective is to investigate the function of Sirtuin 1 (SIRT1) within ARCs. In ARCs tissues and HO-treated lens epithelial cells (LECs), the expression levels of SIRT1 were examined. Senescence-associated β-galactosidase (SA-β-gal) staining was employed to evaluate cellular senescence. The Cell Counting Kit-8 assay was employed to measure viability. A wound healing assay was performed to assess migratory capacity in LECs. Oxidative stress-related indicators were determined by enzyme-linked immunosorbent assay kits. Additionally, the Coxpresdb and GeneCards databases were utilized to identify downstream pathways of SIRT1 in ARCs. The expression levels of protein and mRNA were detected using western blot and real-time quantitative polymerase chain reaction, respectively. The expression of SIRT1 was downregulated in ARCs tissues with an increase in reactive oxygen species. In HO-induced LECs, SIRT1 was downregulated and its overexpression inhibited oxidative stress and cellular senescence while promoting viability and migration. Furthermore, FoxO1/TLR4 pathway was screened out as the key pathway of SIRT1, which was activated in HO-induced LECs senescence. Overexpression of SIRT1 suppressed FoxO1/TLR4 pathway. Further research demonstrated that the activation of FoxO1/TLR4 pathway reversed the inhibitory role of SIRT1 in oxidative stress-induced cellular senescence and the promotion effect of SIRT1 on viability and migration in HO-induced LECs. SIRT1 inhibits oxidative stress-induced cellular senescence and promotes the viability and migration in HO-induced LECs via suppressing FoxO1/TLR4 pathway.
年龄相关性白内障（ARCs）与氧化应激增加和细胞衰老相关。我们的目的是研究沉默调节蛋白1（SIRT1）在年龄相关性白内障中的作用。检测了年龄相关性白内障组织和经过氧化氢（HO）处理的晶状体上皮细胞（LECs）中SIRT1的表达水平。采用衰老相关β-半乳糖苷酶（SA-β-gal）染色评估细胞衰老。采用细胞计数试剂盒-8法检测细胞活力。进行伤口愈合试验以评估晶状体上皮细胞的迁移能力。通过酶联免疫吸附测定试剂盒测定氧化应激相关指标。此外，利用Coxpresdb和GeneCards数据库确定年龄相关性白内障中SIRT1的下游通路。分别采用蛋白质印迹法和实时定量聚合酶链反应检测蛋白质和mRNA的表达水平。在活性氧增加的年龄相关性白内障组织中，SIRT1的表达下调。在HO诱导的晶状体上皮细胞中，SIRT1表达下调，其过表达可抑制氧化应激和细胞衰老，同时促进细胞活力和迁移。此外，筛选出FoxO1/TLR4通路作为SIRT1的关键通路，该通路在HO诱导的晶状体上皮细胞衰老中被激活。SIRT1的过表达抑制了FoxO1/TLR4通路。进一步研究表明，激活FoxO1/TLR4通路可逆转SIRT1对氧化应激诱导的细胞衰老的抑制作用以及SIRT1对HO诱导的晶状体上皮细胞活力和迁移的促进作用。SIRT1通过抑制FoxO1/TLR4通路抑制氧化应激诱导的细胞衰老，并促进HO诱导的晶状体上皮细胞的活力和迁移。
