非诺贝特通过抑制NLRP3炎性小体减轻高糖驱动的人视网膜微血管内皮细胞功能障碍。
Fenofibrate mitigates the dysfunction of high glucose-driven human retinal microvascular endothelial cells by suppressing NLRP3 inflammasome.
作者信息
Shi Yi, Chen Hao-Min, Liu Ai-Hua, Li Xiao-Rong
机构信息
Surgical Retina, Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin Branch of National Clinical Research Center for Ocular Disease, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin 300384, China.
Department of Glaucomatology, Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin Branch of National Clinical Research Center for Ocular Disease, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin 300384, China.
出版信息
Int J Ophthalmol. 2025 May 18;18(5):792-801. doi: 10.18240/ijo.2025.05.04. eCollection 2025.
AIM
To determine the therapeutic benefits of fenofibrate (Feno) on the dysfunction of high glucose (HG)-induced human retinal microvascular endothelial cells (HRMECs) and to elucidate the underlying molecular mechanism.
METHODS
HRMEC dysfunction model was established by 48h glucose (30 mmol/L) treatment and treated with Feno/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activator (Nigericin). Cell viability/apoptosis were assessed by cell counting kit-8 (CCK-8)/terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry assays. Levels of apoptosis- (Bcl-2-associated X protein, Bax/B-cell lymphoma 2, Bcl-2), vascular permeability-(vascular endothelial growth factor, VEGF) and inflammasome activation-related proteins (NLRP3/cleaved caspase-1/apoptosis-associated speck-like protein containing a CARD, ASC), as well as inflammatory factors (interleukin, IL-6/IL-1β/tumor necrosis factor, TNF-α/IL-18) were determined with Western blot/enzyme linked immunosorbent assay (ELISA). Cell permeability/reactive oxygen species (ROS) level/superoxide dismutase (SOD) activity/malondialdehyde (MDA) content were assessed by Evans blue staining/2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe/SOD kit/MDA kit.
RESULTS
HRMEC dysfunction was successfully induced by HG, evidenced by decreased viability (<0.001), increased apoptosis (<0.001), permeability (<0.001), and inflammatory factor levels (<0.001). Feno treatment significantly ameliorated HG-induced HRMEC dysfunction (<0.01). Meanwhile, HG induction increased ROS production (<0.001) and MDA content (<0.001) in HRMECs, while reducing SOD activity (<0.001), indicative of oxidative stress. This was, however, abolished by Feno (<0.05). Moreover, Feno eliminated activation of NLRP3 inflammasomes (<0.05) in HG-induced HRMECs. Strikingly, activation of NLRP3 inflammasomes partially averted the inhibition of Feno on HG-induced HRMEC dysfunction (<0.05).
CONCLUSION
Feno represses oxidative stress and NLRP3 inflammasome activation, consequently alleviating HG-induced HRMEC dysfunction.
目的
确定非诺贝特（Feno）对高糖（HG）诱导的人视网膜微血管内皮细胞（HRMECs）功能障碍的治疗益处，并阐明其潜在的分子机制。
方法
通过48小时葡萄糖（30 mmol/L）处理建立HRMEC功能障碍模型，并用Feno/核苷酸结合寡聚化结构域样受体热蛋白结构域相关蛋白3（NLRP3）炎性小体激活剂（尼日利亚菌素）进行处理。通过细胞计数试剂盒-8（CCK-8）/末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法（TUNEL）染色和流式细胞术检测细胞活力/凋亡。用蛋白质免疫印迹法/酶联免疫吸附测定（ELISA）测定凋亡相关蛋白（Bcl-2相关X蛋白、Bax/B细胞淋巴瘤-2、Bcl-2）、血管通透性相关蛋白（血管内皮生长因子、VEGF）和炎性小体激活相关蛋白（NLRP3/裂解的半胱天冬酶-1/含CARD的凋亡相关斑点样蛋白、ASC）以及炎性因子（白细胞介素，IL-6/IL-1β/肿瘤坏死因子，TNF-α/IL-18）的水平。通过伊文思蓝染色/2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）荧光探针/超氧化物歧化酶（SOD）试剂盒/丙二醛（MDA）试剂盒评估细胞通透性/活性氧（ROS）水平/SOD活性/MDA含量。
结果
HG成功诱导了HRMEC功能障碍，表现为活力降低（<0.001）、凋亡增加（<0.001）、通透性增加（<0.001）和炎性因子水平增加（<0.001）。Feno处理显著改善了HG诱导的HRMEC功能障碍（<0.01）。同时，HG诱导增加了HRMECs中的ROS产生（<0.001）和MDA含量（<0.001），同时降低了SOD活性（<0.001），表明存在氧化应激。然而，Feno消除了这种情况（<0.05）。此外，Feno消除了HG诱导的HRMECs中NLRP3炎性小体的激活（<0.05）。引人注目的是，NLRP3炎性小体的激活部分抵消了Feno对HG诱导的HRMEC功能障碍的抑制作用（<0.05）。
结论
Feno可抑制氧化应激和NLRP3炎性小体激活，从而减轻HG诱导的HRMEC功能障碍。
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