Purpose: To determine whether SIRT1 regulates high glucose (HG)–induced inflammation and cataract formation through modulating TXNIP/NLRP3 inflammasome activation in human lens epithelial cells (HLECs) and rat lenses.

Methods: HG stress from 25 to 150 mM was imposed on HLECs, with treatments using small interfering RNAs (siRNAs) targeting NLRP3, TXNIP, and SIRT1, as well as a lentiviral vector (LV) for SIRT1. Rat lenses were cultivated with HG media, with or without the addition of NLRP3 inhibitor MCC950 or SIRT1 agonist SRT1720. High mannitol groups were applied as the osmotic controls. Real-time PCR, Western blots, and immunofluorescent staining evaluated the mRNA and protein levels of SIRT1, TXNIP, NLRP3, ASC, and IL-1β. Reactive oxygen species (ROS) generation, cell viability, and death were also assessed.

Results: HG stress induced a decline in SIRT1 expression and caused TXNIP/NLRP3 inflammasome activation in a concentration-dependent manner in HLECs, which was not observed in the high mannitol–treated groups. Knocking down NLRP3 or TXNIP inhibited NLRP3 inflammasome-induced IL-1β p17 secretion under HG stress. Transfections of si-SIRT1 and LV-SIRT1 exerted inverse effects on NLRP3 inflammasome activation, suggesting that SIRT1 acts as an upstream regulator of TXNIP/NLRP3 activity. HG stress induced lens opacity and cataract formation in cultivated rat lenses, which was prevented by MCC950 or SRT1720 treatment, with concomitant reductions in ROS production and TXNIP/NLRP3/IL-1β expression levels.

Conclusions: The TXNIP/NLRP3 inflammasome pathway promotes HG-induced inflammation and HLEC pyroptosis, which is negatively regulated by SIRT1. This suggests viable strategies for treating diabetic cataract.

目的：确定 SIRT1 是否通过调节人晶状体上皮细胞 (HLEC) 和大鼠晶状体中的 TXNIP/NLRP3 炎性体激活来调节高葡萄糖 (HG) 诱导的炎症和白内障形成。

方法：对 HLEC 施加 25 至 150 mM 的 HG 压力，使用靶向 NLRP3、TXNIP 和 SIRT1 的小干扰 RNA (siRNA) 以及针对 SIRT1 的慢病毒载体 (LV) 进行处理。用 HG 培养基培养大鼠晶状体，添加或不添加 NLRP3 抑制剂 MCC950 或 SIRT1 激动剂 SRT1720。应用高甘露醇组作为渗透压对照。实时 PCR、蛋白质印迹和免疫荧光染色评估了 SIRT1、TXNIP、NLRP3、ASC 和 IL-1β 的 mRNA 和蛋白质水平。还评估了活性氧 (ROS) 的产生、细胞活力和死亡。

结果：HG 应激诱导 SIRT1 表达下降，并在 HLEC 中以浓度依赖性方式引起 TXNIP/NLRP3 炎性体激活，这在高甘露醇处理组中未观察到。敲低 NLRP3 或 TXNIP 可抑制 HG 胁迫下 NLRP3 炎性体诱导的 IL-1β p17 分泌。si-SIRT1 和 LV-SIRT1 的转染对 NLRP3 炎性体激活产生反向影响，表明 SIRT1 作为 TXNIP/NLRP3 活性的上游调节剂。HG 应激在培养的大鼠晶状体中诱导晶状体混浊和白内障形成，这通过 MCC950 或 SRT1720 处理得到预防，同时 ROS 产生和 TXNIP/NLRP3/IL-1β 表达水平降低。

结论：TXNIP/NLRP3 炎性体通路促进 HG 诱导的炎症和 HLEC 细胞焦亡，这受 SIRT1 负调控。这表明治疗糖尿病性白内障的可行策略。