ENT institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai 200031, China.
Department of Otolaryngology, Head and Neck Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Int J Biol Sci. 2024 Jan 1;20(2):784-800. doi: 10.7150/ijbs.82003. eCollection 2024.
As an anti-tumor drug widely used in the clinic, cisplatin is limited by its ototoxic side effects associated with various factors, including inflammatory responses. Receptor for Advanced Glycation Endproducts (RAGE) recognizes damage-associated molecular patterns (DAMPs) and promotes stress and inflammation. This study intended to determine the potential behavior of the HMGB1/RAGE axis after cisplatin injury and whether it has a protective effect after inhibiting this pathway. We used FPS-ZM1, a RAGE inhibitor, to modulate the axis of HMGB1/RAGE in neonatal mouse cochlear explants and C57BL/6 mice . Apoptosis was identified by Annexin V-FITC/PI assay, Cleaved Caspase-3, and TUNEL staining. Reactive oxygen species (ROS) level was assessed by MitoSOX Red and CellROX Green assay. The expression of proteins associated with the HMGB1/RAGE axis and apoptosis was observed by western blotting. The expression of inflammatory cytokines was evaluated by qPCR. The protective effect of HMGB1/RAGE knockdown was also assessed on cisplatin-induced ototoxicity. These results demonstrated that cisplatin could activate the HMGB1/RAGE pathway in cochlear hair cells and release inflammatory factors. Pretreatment with FPS-ZM1 alleviated cisplatin-induced ototoxicity and . Knocking down HMGB1 and RAGE achieved specific protective effects. Altogether, inhibiting HMGB1/RAGE axis can reverse the increase of ROS accumulation, the activation of apoptosis, and the production of inflammatory reactions after cisplatin injury. FPS-ZM1 could resist the ototoxicity of cisplatin by suppressing the HMGB1/RAGE signal pathway, and it may be considered the new otoprotective potential strategy for hearing loss.
作为一种在临床上广泛应用的抗肿瘤药物，顺铂受到其耳毒性副作用的限制，这些副作用与多种因素有关，包括炎症反应。晚期糖基化终产物受体（RAGE）识别损伤相关分子模式（DAMPs），并促进应激和炎症反应。本研究旨在确定顺铂损伤后 HMGB1/RAGE 轴的潜在行为，以及抑制该途径后是否具有保护作用。我们使用 RAGE 抑制剂 FPS-ZM1 来调节新生鼠耳蜗外植体和 C57BL/6 小鼠中 HMGB1/RAGE 轴。通过 Annexin V-FITC/PI 检测、Cleaved Caspase-3 和 TUNEL 染色鉴定细胞凋亡。通过 MitoSOX Red 和 CellROX Green 检测评估活性氧（ROS）水平。通过 Western blot 观察与 HMGB1/RAGE 轴和凋亡相关的蛋白表达。通过 qPCR 评估炎症细胞因子的表达。还评估了 HMGB1/RAGE 敲低对顺铂诱导的耳毒性的保护作用。结果表明，顺铂可激活耳蜗毛细胞中的 HMGB1/RAGE 途径并释放炎症因子。FPS-ZM1 预处理可减轻顺铂引起的耳毒性。敲低 HMGB1 和 RAGE 可实现特异性保护作用。总之，抑制 HMGB1/RAGE 轴可以逆转顺铂损伤后 ROS 积累、凋亡激活和炎症反应的增加。FPS-ZM1 通过抑制 HMGB1/RAGE 信号通路抵抗顺铂的耳毒性，可能被视为听力损失的新的潜在的耳保护策略。
