Interdisciplinary Graduate Program in Advanced Convergence Technology & Science, Jeju National University, Jeju, Jeju Self-Governing Province, Republic of Korea.
Department of Urology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
Pharmacol Rep. 2022 Oct;74(5):1041-1053. doi: 10.1007/s43440-022-00403-x. Epub 2022 Aug 22.
Kidney ischemia reperfusion injury (IRI) is characterized by tubular cell death. DNA double-strand breaks is one of the major sources of tubular cell death induced by IRI. 2-Mercaptoethanol (2-ME) is protective against DNA double-strand breaks derived from calf thymus and bovine embryo. Here, we sought to determine whether treatment with 2-ME attenuated DNA double-strand breaks, resulting in reduced kidney dysfunction and structural damage in IRI.
Kidney IRI or sham-operation in mice was carried out. The mice were treated with 2-ME, Ras-selective lethal 3, or vehicle. Kidney function, tubular injury, DNA damage, antioxidant enzyme expression, and DNA damage response (DDR) kinases activation were assessed.
Treatment with 2-ME significantly attenuated kidney dysfunction, tubular injury, and DNA double-strand breaks after IRI. Among DDR kinases, IRI induced phosphorylation of ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and Rad3 related (ATR), but IRI reduced phosphorylation of other DDR kinases including ataxia telangiectasia and Rad3 related, checkpoint kinase 1 (Chk1), Chk2, and Chinese hamster cells 1 (XRCC1). Treatment with 2-ME enhanced phosphorylation of ATM and ATM-mediated effector kinases in IRI-subjected kidneys, suggesting that 2-ME activates ATM-mediated DDR signaling pathway. Furthermore, 2-ME dramatically upregulated glutathione peroxidase 4 (GPX4) in IRI-subjected kidneys. Inhibition of GPX4 augmented adverse IRI consequences including kidney dysfunction, tubular injury, DNA double-strand breaks, and inactivation of ATM-mediated DDR signaling pathway after IRI in 2-ME-treated kidneys.
We have demonstrated that exogenous 2-ME protects against DNA double-strand breaks after kidney IRI through GPX4 upregulation and ATM activation.
肾缺血再灌注损伤（IRI）的特征是肾小管细胞死亡。DNA 双链断裂是 IRI 诱导的肾小管细胞死亡的主要来源之一。2-巯基乙醇（2-ME）可防止小牛胸腺和牛胚胎来源的 DNA 双链断裂。在这里，我们试图确定 2-ME 治疗是否能减轻 DNA 双链断裂，从而减少 IRI 引起的肾功能障碍和结构损伤。
在小鼠中进行肾 IRI 或假手术。用 2-ME、Ras 选择性致死 3 或载体处理小鼠。评估肾功能、肾小管损伤、DNA 损伤、抗氧化酶表达和 DNA 损伤反应（DDR）激酶激活。
2-ME 治疗可显著减轻 IRI 后肾功能障碍、肾小管损伤和 DNA 双链断裂。在 DDR 激酶中，IRI 诱导共济失调毛细血管扩张突变（ATM）和共济失调毛细血管扩张和 Rad3 相关（ATR）的磷酸化，但 IRI 降低了其他 DDR 激酶如共济失调毛细血管扩张和 Rad3 相关、检查点激酶 1（Chk1）、Chk2 和中国仓鼠细胞 1（XRCC1）的磷酸化。2-ME 治疗增强了 IRI 肾脏中 ATM 和 ATM 介导的效应激酶的磷酸化，表明 2-ME 激活了 ATM 介导的 DDR 信号通路。此外，2-ME 在 IRI 肾脏中显著上调了谷胱甘肽过氧化物酶 4（GPX4）。在 2-ME 处理的肾脏中，抑制 GPX4 后，IRI 引起的肾功能障碍、肾小管损伤、DNA 双链断裂和 ATM 介导的 DDR 信号通路失活等不良后果加剧。
我们已经证明，外源性 2-ME 通过上调 GPX4 和激活 ATM 来保护肾 IRI 后的 DNA 双链断裂。
