Department of Anesthesiology, ZhuJiang Hospital, Southern Medical University, Guangdong Province, China.
State Key Laboratory of Pharmaceutical Biotechnology and Department of Anesthesiology, University of Hong Kong, Hong Kong.
Aging (Albany NY). 2021 Jan 20;13(3):4274-4290. doi: 10.18632/aging.202390.
Bupivacaine has been widely used in clinical Anesthesia, but its neurotoxicity has been frequently reported, implicating cellular oxidative DNA damage as the major underlying mechanism. However, the mechanism underlying bupivacaine-induced oxidative DNA damage is unknown. We, thus, exposed SH-SY5Y cells to 1.5mM bupivacaine to induce neurotoxicity. Then, iTRAQ proteomic analysis was used to explore the repair of neuronal oxidative DNA damage. By analyzing the STRING version 11.0 database, the bioinformatics relationship between key repair enzymes was tracked. Subsequently, immunofluorescence co-localization and immunoprecipitation were used to investigate the interaction between key repair enzymes. The iTRAQ showed that Poly [ADP-ribose] polymerase 1 (PARP-1) from the base excision repair pathway participated closely in the repair of oxidative DNA damage induced by bupivacaine, and inhibition of PARP-1 expression significantly aggravated bupivacaine-induced DNA damage and apoptosis. Interestingly, this study showed that there were interactions and co-expression between PARP-1 and XPD (xeroderma pigmentosum D), another key protein of the nucleic acid excision repair pathway. After inhibiting XPD, PARP-1 expression was significantly reduced. However, simultaneous inhibition of both XPD and PARP-1 did not further increase DNA damage. It is concluded that PARP-1 may repair bupivacaine-induced oxidative DNA damage through XPD-mediated interactions.
布比卡因被广泛应用于临床麻醉,但它的神经毒性已被频繁报道,提示细胞氧化 DNA 损伤是其主要的潜在机制。然而,布比卡因诱导氧化 DNA 损伤的机制尚不清楚。我们因此将 SH-SY5Y 细胞暴露于 1.5mM 布比卡因中,以诱导神经毒性。然后,我们使用 iTRAQ 蛋白质组学分析来探索神经元氧化 DNA 损伤的修复。通过分析 STRING 版本 11.0 数据库,追踪了关键修复酶之间的生物信息学关系。随后,我们通过免疫荧光共定位和免疫沉淀实验来研究关键修复酶之间的相互作用。iTRAQ 显示,碱基切除修复途径中的聚 ADP-核糖聚合酶 1(PARP-1)密切参与布比卡因诱导的氧化 DNA 损伤的修复,而 PARP-1 表达的抑制显著加重了布比卡因诱导的 DNA 损伤和细胞凋亡。有趣的是,本研究表明 PARP-1 与核酸切除修复途径中的另一个关键蛋白 XPD(着色性干皮病 D)之间存在相互作用和共表达。抑制 XPD 后,PARP-1 的表达显著降低。然而,同时抑制 XPD 和 PARP-1 并没有进一步增加 DNA 损伤。综上所述,PARP-1 可能通过 XPD 介导的相互作用来修复布比卡因诱导的氧化 DNA 损伤。