Choi Seung Gyu, Shin Juhee, Lee Ka Young, Park Hyewon, Kim Song I, Yi Yoon Young, Kim Dong Woon, Song Hee-Jung, Shin Hyo Jung
Department of Medical Science, Chungnam National University, Daejeon, Republic of Korea.
Department of Anatomy and Cell Biology, Chungnam National University, Daejeon, Republic of Korea.
Glia. 2023 May;71(5):1294-1310. doi: 10.1002/glia.24339. Epub 2023 Jan 18.
PTEN-induced kinase 1 (PINK1) is a well-known critical marker in the pathway for mitophagy regulation as well as mitochondrial dysfunction. Evidence suggests that mitochondrial dynamics and mitophagy flux play an important role in the development of brain damage from stroke pathogenesis. In this study, we propose a treatment strategy using nanoparticles that can control PINK1. We used a murine photothrombotic ischemic stroke (PTS) model in which clogging of blood vessels is induced with Rose Bengal (RB) to cause brain damage. We targeted PINK1 with poly(lactic-co-glycolic acid) (PLGA)-based nanoparticles loaded with PINK1 siRNA (PINK1 NPs). After characterizing siRNA loading in the nanoparticles, we assessed the efficacy of PINK1 NPs in mice with PTS using immunohistochemistry, 1% 2,3,5-triphenyltetrazolium chloride staining, measurement of motor dysfunction, and Western blot. PINK1 was highly expressed in microglia 24 h after PTS induction. PINK1 siRNA treatment increased phagocytic activity, migration, and expression of an anti-inflammatory state in microglia. In addition, the PLGA nanoparticles were selectively taken up by microglia and specifically regulated PINK1 expression in those cells. Treatment with PINK1 NPs prior to stroke induction reduced expression of mitophagy-inducing factors, infarct volume, and motor dysfunction in mice with photothrombotic ischemia. Experiments with PINK1-knockout mice and microglia depletion with PLX3397 confirmed a decrease in stroke-induced infarct volume and behavioral dysfunction. Application of nanoparticles for PINK1 inhibition attenuates RB-induced photothrombotic ischemic injury by inhibiting microglia responses, suggesting that a nanomedical approach targeting the PINK1 pathway may provide a therapeutic avenue for stroke treatment.
PTEN诱导激酶1（PINK1）是线粒体自噬调节途径以及线粒体功能障碍中一个众所周知的关键标志物。有证据表明，线粒体动力学和线粒体自噬通量在中风发病机制导致的脑损伤发展中起重要作用。在本研究中，我们提出了一种使用可调控PINK1的纳米颗粒的治疗策略。我们使用了一种小鼠光血栓性缺血性中风（PTS）模型，其中用孟加拉玫瑰红（RB）诱导血管阻塞以导致脑损伤。我们用负载有PINK1小干扰RNA（PINK1 NPs）的聚乳酸-羟基乙酸共聚物（PLGA）基纳米颗粒靶向PINK1。在表征纳米颗粒中的小干扰RNA负载情况后，我们使用免疫组织化学、1% 2,3,5-三苯基氯化四氮唑染色、运动功能障碍测量和蛋白质免疫印迹法评估了PINK1 NPs对PTS小鼠的疗效。PTS诱导后24小时，PINK1在小胶质细胞中高表达。PINK1小干扰RNA处理增加了小胶质细胞的吞噬活性、迁移以及抗炎状态的表达。此外，PLGA纳米颗粒被小胶质细胞选择性摄取，并特异性调节这些细胞中PINK1的表达。在中风诱导前用PINK1 NPs处理可降低光血栓性缺血小鼠中自噬诱导因子的表达、梗死体积和运动功能障碍。用PINK1基因敲除小鼠和用PLX3397清除小胶质细胞的实验证实了中风诱导的梗死体积和行为功能障碍的减少。应用纳米颗粒抑制PINK1可通过抑制小胶质细胞反应减轻RB诱导的光血栓性缺血损伤，这表明靶向PINK1途径的纳米医学方法可能为中风治疗提供一条治疗途径。
