Department of Toxicological Biochemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Medyczna 9, 30-688 Kraków, Poland.
Department of Neurology, Faculty of Medicine, Jagiellonian University Medical College, Botaniczna 3, 31-503 Kraków, Poland.
Int J Mol Sci. 2021 Jul 22;22(15):7816. doi: 10.3390/ijms22157816.
Ischemic stroke is the third leading cause of death in the world, which accounts for almost 12% of the total deaths worldwide. Despite decades of research, the available and effective pharmacotherapy is limited. Some evidence underlines the beneficial properties of hydrogen sulfide (HS) donors, such as NaSH, in an animal model of brain ischemia and in in vitro research; however, these data are ambiguous. This study was undertaken to verify the neuroprotective activity of AP39, a slow-releasing mitochondria-targeted HS delivery molecule. We administered AP39 for 7 days prior to ischemia onset, and the potential to induce brain tolerance to ischemia was verified. To do this, we used the rat model of 90-min middle cerebral artery occlusion (MCAO) and used LC-MS/MS, RT-PCR, Luminex assays, Western blot and immunofluorescent double-staining to determine the absolute HS levels, inflammatory markers, neurotrophic factor signaling pathways and apoptosis marker in the ipsilateral frontal cortex, hippocampus and in the dorsal striatum 24 h after ischemia onset. AP39 (50 nmol/kg) reduced the infarct volume, neurological deficit and reduced the microglia marker (Iba1) expression. AP39 also exerted prominent anti-inflammatory activity in reducing the release of Il-1β, Il-6 and TNFα in brain areas particularly affected by ischemia. Furthermore, AP39 enhanced the pro-survival pathways of neurotrophic factors BDNF-TrkB and NGF-TrkA and reduced the proapoptotic proNGF-p75-sortilin pathway activity. These changes corresponded with reduced levels of cleaved caspase 3. Altogether, AP39 treatment induced adaptative changes within the brain and, by that, developed brain tolerance to ischemia.
缺血性中风是世界上第三大致死原因，占全球总死亡人数的近 12%。尽管经过几十年的研究，可用的有效药物治疗方法仍然有限。一些证据强调了硫化氢（HS）供体，如 NaSH，在脑缺血动物模型和体外研究中的有益特性；然而，这些数据存在歧义。本研究旨在验证 AP39 的神经保护活性，AP39 是一种缓慢释放的靶向线粒体的 HS 递药分子。我们在缺血发作前给予 AP397 天，并验证其诱导脑对缺血的耐受能力。为此，我们使用 90 分钟大脑中动脉闭塞（MCAO）大鼠模型，并用 LC-MS/MS、RT-PCR、Luminex 检测、Western blot 和免疫荧光双重染色来确定同侧额皮质、海马体和背侧纹状体中的绝对 HS 水平、炎症标志物、神经营养因子信号通路和凋亡标志物在缺血发作后 24 小时。AP39（50 nmol/kg）减少了梗死体积、神经功能缺损和小胶质细胞标志物（Iba1）的表达。AP39 还通过减少受缺血影响特别严重的脑区中 Il-1β、Il-6 和 TNFα 的释放发挥了显著的抗炎活性。此外，AP39 增强了神经营养因子 BDNF-TrkB 和 NGF-TrkA 的促生存途径，并降低了促凋亡 proNGF-p75-sortilin 途径的活性。这些变化与 cleaved caspase 3 水平的降低相对应。总之，AP39 治疗诱导了大脑内的适应性变化，并由此产生了脑对缺血的耐受能力。
