Li Rong, Chen Lin, Yao Guo-Min, Yan Hong-Lin, Wang Li
Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an 710077, Shaanxi Province, China.
Department of Pharmacy, the First Affiliated Hospital of Xi'an Medical University, Xi'an 710077, Shaanxi Province, China.
Int J Ophthalmol. 2021 Jan 18;14(1):42-49. doi: 10.18240/ijo.2021.01.06. eCollection 2021.
To investigate the effects of quercetin on diabetic retinopathy (DR) and its association with nucleotide-binding oligomerization domain-like receptors 3 (NLRP3) inflammasome and autophagy using retinal endothelial cell as an experimental model.
Human retinal microvascular endothelial cells (HRMECs) were cultured and assigned into the control group, high-glucose (HG) group, and HG+different concentrations of quercetin groups. Cellular viability, migration, and tube formation in these groups was detected by MTT, transwell and matrigel assay, respectively. Expressions of NLRP3, apoptosis-associated speck-like protein (ASC), cysteiny aspartate-specific protease-1 (Caspase-1) as well as microtubule-related protein 1 light chain 3 (LC3) and Beclin-1 were detected by Western blotting. Expressions of IL-1β and IL-18 were detected by ELISA and cellular autophagy was detected by Cyto-ID autophagy detection kit.
Under an HG condition, the viability, migration, tube formation of HRMECs, and the protein expressions of NLRP3, ASC, Caspase-1, IL-1β, IL-18, LC3, and Beclin-1 as well as autophagy were all increased. Quercetin inhibited angiogenesis of HRMECs as well as the expressions of NLRP3, ASC, Caspase-1, IL-1β, IL-18, LC3, Beclin-1, and autophagy of HRMECs under a HG condition. The inhibitory effects of quercetin on angiogenesis, NLRP3 inflammasome and autophagy increased with the increase of its concentration.
The therapeutic potential of quercetin in retinal neovascularization of DR, and inhibition of NLRP3 inflammasome and autophagy signaling pathway may be involved.
以视网膜内皮细胞为实验模型,研究槲皮素对糖尿病视网膜病变(DR)的影响及其与核苷酸结合寡聚化结构域样受体3(NLRP3)炎性小体和自噬的关系。
培养人视网膜微血管内皮细胞(HRMECs),分为对照组、高糖(HG)组和HG+不同浓度槲皮素组。分别采用MTT法、Transwell法和基质胶实验检测各组细胞活力、迁移能力和成管能力。通过蛋白质免疫印迹法检测NLRP3、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸特异性蛋白酶-1(Caspase-1)以及微管相关蛋白1轻链3(LC3)和Beclin-1的表达。采用酶联免疫吸附测定(ELISA)法检测白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)的表达,并用Cyto-ID自噬检测试剂盒检测细胞自噬。
在HG条件下,HRMECs的活力、迁移能力、成管能力以及NLRP3、ASC、Caspase-1、IL-1β、IL-18、LC3、Beclin-1的蛋白表达和自噬均增加。槲皮素抑制HG条件下HRMECs的血管生成以及NLRP3、ASC、Caspase-1、IL-1β、IL-18、LC3、Beclin-1的表达和HRMECs的自噬。槲皮素对血管生成、NLRP3炎性小体和自噬的抑制作用随其浓度增加而增强。
槲皮素在DR视网膜新生血管形成中具有治疗潜力,可能涉及抑制NLRP3炎性小体和自噬信号通路。