Sun Haiyan, Zhao Huahui, Yan Zhipeng, Liu Xiaokun, Yin Pengfei, Zhang Jun
Ophthalmology Department, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, P.R. China.
Exp Ther Med. 2021 Aug;22(2):833. doi: 10.3892/etm.2021.10265. Epub 2021 Jun 3.
Diabetic retinopathy (DR) is a major complication of diabetes mellitus that may cause severe visual impairment. It has been reported that the levels of nesfatin-1 in the serum and vitreous humor were negatively correlated with DR; however, its role in DR has not been fully elucidated. Therefore, the present study was performed to investigate the effect of nesfatin-1 on high glucose-treated human retinal epithelial cells (ARPE-19) and explore the underlying mechanism. The effects of nesfatin-1 on cell viability, inflammation, oxidative stress and apoptosis were examined under high glucose conditions. The Cell Counting Kit-8 assay was used to determine cell viability. The levels of inflammatory cytokines were evaluated using ELISA kits. The reactive oxygen species and malondialdehyde content was estimated using commercial assay kits. Flow cytometry was performed to detect apoptotic cells and western blot analysis was employed to evaluate the expression of apoptosis-associated proteins. Moreover, the levels of NF-κB, NACHT, LRR and PYD domains-containing protein 3 (NLRP3) and high-mobility group protein B1 (HMGB1) were determined via western blot analysis. The results revealed that nesfatin-1 enhanced cell viability and suppressed inflammation, oxidative stress and apoptosis in the presence of high glucose concentration. Moreover, the activation of the NF-κB/NLRP3 inflammasome signaling and the expression of HMGB1 were inhibited by nesfatin-1. Furthermore, HMGB1 overexpression partially abrogated the inactivation of the NF-κB/NLRP3 inflammasome pathway caused by nesfatin-1. Taken together, these findings demonstrated that nesfatin-1 inhibited the activation of the NF-κB/NLRP3 inflammasome signaling via modulating HMGB1 and exerted a protective effect on ARPE-19 cells against high glucose-induced inflammation, oxidative stress and apoptosis.
糖尿病性视网膜病变(DR)是糖尿病的一种主要并发症,可能导致严重的视力损害。据报道,血清和玻璃体液中nesfatin-1的水平与DR呈负相关;然而,其在DR中的作用尚未完全阐明。因此,本研究旨在探讨nesfatin-1对高糖处理的人视网膜上皮细胞(ARPE-19)的影响,并探索其潜在机制。在高糖条件下检测了nesfatin-1对细胞活力、炎症、氧化应激和细胞凋亡的影响。使用细胞计数试剂盒-8检测法测定细胞活力。使用酶联免疫吸附测定试剂盒评估炎性细胞因子的水平。使用商业检测试剂盒估计活性氧和丙二醛含量。进行流式细胞术检测凋亡细胞,并采用蛋白质免疫印迹分析评估凋亡相关蛋白的表达。此外,通过蛋白质免疫印迹分析测定核因子κB(NF-κB)、含NACHT、亮氨酸重复序列和PYD结构域蛋白3(NLRP3)和高迁移率族蛋白B1(HMGB1)的水平。结果显示,在高糖浓度存在的情况下,nesfatin-1增强了细胞活力,抑制了炎症、氧化应激和细胞凋亡。此外,nesfatin-1抑制了NF-κB/NLRP3炎性小体信号的激活以及HMGB1的表达。此外,HMGB1过表达部分消除了nesfatin-1引起的NF-κB/NLRP3炎性小体途径的失活。综上所述,这些发现表明,nesfatin-1通过调节HMGB1抑制NF-κB/NLRP3炎性小体信号的激活,并对ARPE-19细胞免受高糖诱导的炎症、氧化应激和细胞凋亡发挥保护作用。