The Department of Anesthesiology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, China.
Biol Res. 2024 Oct 5;57(1):71. doi: 10.1186/s40659-024-00547-5.
Stroke is a leading cause of death worldwide, with oxidative stress and calcium overload playing significant roles in the pathophysiology of the disease. Ozone, renowned for its potent antioxidant properties, is commonly employed as an adjuvant therapy in clinical settings. Nevertheless, it remains unclear whether ozone therapy on parthanatos in cerebral ischemia-reperfusion injury (CIRI). This study aims to investigate the impact of ozone therapy on reducing parthanatos during CIRI and to elucidate the underlying mechanism.
Hydrogen peroxide (HO) was utilized to mimic the generation of reactive oxygen species (ROS) in SH-SY5Y cell reperfusion injury in vitro, and an in vivo ischemic stroke model was established. Ozone saline was introduced for co-culture or intravenously administered to mice. Apoptosis and oxidative stress were assessed using flow cytometry and immunofluorescence. Western blotting was utilized to examine the expression of parthanatos signature proteins. The mechanism by which ozone inhibits parthanatos was elucidated through inhibiting PPARg or Nrf2 activity.
The findings demonstrated that ozone mitigated HO-induced parthanatos by either upregulating nuclear factor erythroid 2-related factor 2 (Nrf2) or activating peroxisome proliferator-activated receptorg (PPARg). Furthermore, through the use of calcium chelators and ROS inhibitors, it was discovered that ROS directly induced parthanatos and facilitated intracellular calcium elevation. Notably, a malignant feedback loop between ROS and calcium was identified, further amplifying the induction of parthanatos. Ozone therapy exhibited its efficacy by increasing PPARg activity or enhancing the Nrf2 translation, thereby inhibiting ROS production induced by HO. Concurrently, our study demonstrated that ozone treatment markedly inhibited parthanatos in stroke-afflicted mice. Additionally, ozone therapy demonstrated significant neuroprotective effects on cortical neurons, effectively suppressing parthanatos.
These findings contribute valuable insights into the potential of ozone therapy as a therapeutic strategy for reducing parthanatos during CIRI, highlighting its impact on key molecular pathways associated with oxidative stress and calcium regulation.
脑卒中是全球范围内的主要致死病因，氧化应激和钙超载在疾病的病理生理学中发挥着重要作用。臭氧以其强大的抗氧化特性而闻名，常被用作临床辅助治疗。然而，臭氧治疗是否能减轻脑缺血再灌注损伤（CIRI）中的细胞调亡途径仍不清楚。本研究旨在探讨臭氧治疗减轻 CIRI 中细胞调亡途径的作用，并阐明其潜在机制。
采用过氧化氢（HO）模拟体外 SH-SY5Y 细胞再灌注损伤中活性氧（ROS）的产生，建立体内缺血性脑卒中模型。臭氧生理盐水用于共培养或静脉注射到小鼠体内。采用流式细胞术和免疫荧光法评估细胞凋亡和氧化应激。采用 Western blot 检测细胞调亡途径特征蛋白的表达。通过抑制过氧化物酶体增殖物激活受体γ（PPARg）或核因子红细胞 2 相关因子 2（Nrf2）活性，阐明臭氧抑制细胞调亡途径的机制。
结果表明，臭氧通过上调核因子红细胞 2 相关因子 2（Nrf2）或激活过氧化物酶体增殖物激活受体γ（PPARg）减轻 HO 诱导的细胞调亡途径。此外，通过使用钙螯合剂和 ROS 抑制剂，发现 ROS 直接诱导细胞调亡途径，并促进细胞内钙升高。值得注意的是，ROS 和钙之间存在一个恶性反馈环，进一步放大了细胞调亡途径的诱导。臭氧治疗通过增加 PPARg 活性或增强 Nrf2 翻译来发挥其疗效，从而抑制 HO 诱导的 ROS 产生。同时，我们的研究表明，臭氧治疗显著抑制了脑卒中小鼠的细胞调亡途径。此外，臭氧治疗对皮质神经元具有显著的神经保护作用，有效抑制细胞调亡途径。
这些发现为臭氧治疗作为减轻 CIRI 中细胞调亡途径的治疗策略提供了有价值的见解，强调了其对与氧化应激和钙调节相关的关键分子途径的影响。
