Department of Ophthalmology, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Taizhou School of Clinical Medicine, Nanjing Medical University, Taizhou, Jiangsu, China.
Department of Ophthalmology, Lixiang Eye Hospital of Soochow University, Suzhou, Jiangsu, China.
Curr Eye Res. 2024 Dec;49(12):1285-1294. doi: 10.1080/02713683.2024.2378037. Epub 2024 Aug 5.
Diabetic retinopathy (DR) is one of the most severe and common complications caused by diabetic mellites. Inhibiting NLRP3 inflammasome activation displays a crucial therapeutic value in DR. Studies have shown that KCNQ1OT1 plays a critical role in regulating NLRP3 inflammasome activation and participates in the pathogenesis of diabetic complications. The present study aims to explore the role, and the potential mechanism of KCNQ1OT1 in regulating the activation of NLRP3 inflammasome in DR.
qRT-PCR was used to detect the expression of KCNQ1OT1, miR-17-5p, TXNIP, NLRP3, ASC, caspase-1 and IL-1β. Western blot was performed to detect the expression of NLRP3, ASC, caspase-1, IL-1β and TXNIP. Immunohistochemistry and immunostaining were performed to detect the expression of caspase-1. The levels of the inflammatory cytokine IL-1β were determined by ELISA assay. FISH was used to detect the subcellular localisation of KCNQ1OT1. Bioinformatic analysis, luciferase reporter assay and studies were performed to elucidate the mechanism of KCNQ1OT1-mediated dysfunction.
The expression of KCNQ1OT1 and the activation of NLRP3 inflammasome were increased in experimental DR models. KCNQ1OT1 knockdown alleviated NLRP3 inflammasome-associated molecules expression. In addition, KCNQ1OT1 was found to be localized mainly in the cytoplasm of Müller cells and facilitated TXNIP expression by acting as a miR-17-5p sponge. KCNQ1OT1 promoted the activation of NLRP3 inflammasome through miR-17-5p/TXNIP axis.
In conclusion, it was found in this study that KCNQ1OT1 promoted the activation of NLRP3 inflammasome both and , which was mediated by miR-17-5p/TXNIP axis. KCNQ1OT1 might be an effective interference target for the prevention and treatment of DR.
糖尿病视网膜病变（DR）是糖尿病最严重和常见的并发症之一。抑制 NLRP3 炎性小体的激活在 DR 中具有重要的治疗价值。研究表明，KCNQ1OT1 在调节 NLRP3 炎性小体激活中起关键作用，并参与糖尿病并发症的发病机制。本研究旨在探讨 KCNQ1OT1 在调节 DR 中 NLRP3 炎性小体激活中的作用及其潜在机制。
采用 qRT-PCR 检测 KCNQ1OT1、miR-17-5p、TXNIP、NLRP3、ASC、caspase-1 和 IL-1β 的表达。采用 Western blot 检测 NLRP3、ASC、caspase-1、IL-1β 和 TXNIP 的表达。采用免疫组化和免疫染色检测 caspase-1 的表达。采用 ELISA 法检测炎性细胞因子 IL-1β 的水平。采用 FISH 检测 KCNQ1OT1 的亚细胞定位。通过生物信息学分析、荧光素酶报告基因检测和实验研究阐明了 KCNQ1OT1 介导的功能障碍的机制。
在实验性 DR 模型中，KCNQ1OT1 的表达和 NLRP3 炎性小体的激活增加。KCNQ1OT1 敲低减轻了 NLRP3 炎性小体相关分子的表达。此外，发现 KCNQ1OT1 主要定位于 Müller 细胞的细胞质中，并通过作为 miR-17-5p 的海绵发挥作用来促进 TXNIP 的表达。KCNQ1OT1 通过 miR-17-5p/TXNIP 轴促进 NLRP3 炎性小体的激活。
综上所述，本研究发现 KCNQ1OT1 通过 miR-17-5p/TXNIP 轴促进 NLRP3 炎性小体的激活，从而促进 NLRP3 炎性小体的激活。KCNQ1OT1 可能是预防和治疗 DR 的有效干预靶点。
