Cao Xin, Di Yue, Tian Ya-Jing, Huang Xiao-Bo, Zhou Yue, Zhang Dong-Mei, Song Yu
Department of Ophthalmology, The Second Affiliated Hospital of Nantong University, No. 666, Shengli Road, Nantong, 226001, Jiangsu Province, People's Republic of China.
Int Ophthalmol. 2025 Mar 18;45(1):108. doi: 10.1007/s10792-025-03458-w.
To determine the impact of sodium butyrate on the activation of the reactive oxygen species (ROS)/nuclear factor kappa B (NF-κB)/NLR family pyrin domain containing 3 (NLRP3) signaling pathway and angiogenesis in human retinal microvascular endothelial cells (HRMECs) caused by high glucose (HG).
HRMECs were grown for 24 h or 72 h in HG solution (30 mmol/L D-glucose) with 5 mM NaB. Using Cell Counting Kit-8, the effects of HG and NaB levels on the viability of HRMECs were examined. Using various kits, intracellular ROS levels, lactate dehydrogenase (LDH), and Malondialdehyde (MDA) in cell supernatants were measured. Western blot, Immunofluorescence, and Real-time quantitative polymerase chain reaction were employed to quantify protein and messenger RNA expression. Using wound-healing and tube formation tests, the migratory proficiency and angiogenesis of HRMECs were evaluated.
NaB demonstrated a reduction in ROS production, as well as the release of LDH and MDA in HG-induced HRMECs. Additionally, NaB led to a decrease in protein expression of phosphorylation of NF-κB, NLRP3, Caspase 1, interleukin, vascular cell adhesion molecule-1 and intercellular adhesion molecule-1. The impact of HG on zonula occluden-1, a tight junction protein, was attenuated by NaB. Furthermore, NaB inhibited the migration and tube formation of HRMECs partly by ROS/NF-κB/NLRP3 pathway.
NaB suppresses the activation of ROS/NF-κB/NLRP3 signaling pathway and angiogenesis in HRMECs induced by HG.
确定丁酸钠对高糖（HG）诱导的人视网膜微血管内皮细胞（HRMECs）中活性氧（ROS）/核因子κB（NF-κB）/含NLR家族吡咯结构域3（NLRP3）信号通路激活及血管生成的影响。
将HRMECs在含5 mM NaB的HG溶液（30 mmol/L D-葡萄糖）中培养24小时或72小时。使用细胞计数试剂盒-8检测HG和NaB水平对HRMECs活力的影响。使用各种试剂盒测量细胞内ROS水平、细胞上清液中的乳酸脱氢酶（LDH）和丙二醛（MDA）。采用蛋白质印迹法、免疫荧光法和实时定量聚合酶链反应来定量蛋白质和信使核糖核酸表达。使用伤口愈合和管腔形成试验评估HRMECs的迁移能力和血管生成。
NaB可降低HG诱导的HRMECs中ROS的产生，以及LDH和MDA的释放。此外，NaB导致NF-κB、NLRP3、半胱天冬酶1、白细胞介素、血管细胞黏附分子-1和细胞间黏附分子-1磷酸化的蛋白质表达降低。NaB减弱了HG对紧密连接蛋白闭合蛋白-1的影响。此外，NaB部分通过ROS/NF-κB/NLRP3途径抑制HRMECs的迁移和管腔形成。
NaB抑制HG诱导的HRMECs中ROS/NF-κB/NLRP3信号通路的激活及血管生成。
