Hu Na, Li Yunfeng, Zhang Guohong, Wang Wei, An Liping, An Ran, Liu Yu
Department of Biochemistry and Biology, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei Province, China; Hebei Key Laboratory of Chinese Medicine Research on Cardio- Cerebrovascular Disease, Shijiazhuang, Hebei Province, China.
Biomol Biomed. 2025 Apr 26;25(6):1322-1334. doi: 10.17305/bb.2024.11415.
Inhibiting ferroptosis represents a promising strategy for managing neuronal injury caused by intracerebral hemorrhage (ICH). Platelet factor 4 (PF4), a chemokine with diverse biological functions, has an unclear role in ICH and its impact on neuronal ferroptosis. To investigate this, a hemin-induced injury model was established in PC12 cells in vitro, and an ICH model was created in vivo using IV collagenase injection. Hemin-treated PC12 cells were co-cultured with recombinant mouse PF4 (Rm-PF4) protein to examine the effects of PF4 on ferroptosis. Additionally, Rm-PF4 was administered intraperitoneally to ICH mice, and its influence on neurological dysfunction, brain edema, and neuronal ferroptosis was evaluated. Western blot analysis was employed to assess PF4 levels, CXCR3/phosphatidylinositol 3-kinase (PI3K)/AKT/nuclear factor erythroid-2-related factor 2 (Nrf2) pathway activation, and ferroptosis-related protein expression. PF4 levels were found to be reduced in both perihematomal brain tissues of ICH mice and hemin-treated PC12 cells. Treatment with Rm-PF4 decreased ferrous ion, malondialdehyde (MDA), and reactive oxygen species (ROS) levels, effectively inhibiting ferroptosis in PC12 cells. Furthermore, Rm-PF4 administration alleviated neurological dysfunction, neuronal damage, and brain edema while suppressing neuronal ferroptosis in ICH mice. Mechanistically, Rm-PF4 activated the CXCR3/PI3K/AKT/Nrf2 pathway, and this protective effect was diminished by a CXCR3 antagonist in both ICH mice and hemin-treated PC12 cells. In conclusion, PF4 mitigates ICH-induced neuronal ferroptosis in mouse models and PC12 cells by activating the CXCR3/PI3K/AKT/Nrf2 pathway.
抑制铁死亡是治疗脑出血（ICH）所致神经元损伤的一种有前景的策略。血小板因子4（PF4）是一种具有多种生物学功能的趋化因子，其在ICH中的作用及其对神经元铁死亡的影响尚不清楚。为了研究这一点，体外在PC12细胞中建立了血红素诱导的损伤模型，并通过静脉注射胶原酶在体内创建了ICH模型。将经血红素处理的PC12细胞与重组小鼠PF4（Rm-PF4）蛋白共培养，以研究PF4对铁死亡的影响。此外，将Rm-PF4腹腔注射给ICH小鼠，并评估其对神经功能障碍、脑水肿和神经元铁死亡的影响。采用蛋白质印迹分析评估PF4水平、CXCR3/磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）/核因子红细胞2相关因子2（Nrf2）通路激活情况以及铁死亡相关蛋白表达。发现ICH小鼠血肿周围脑组织和经血红素处理的PC12细胞中PF4水平均降低。用Rm-PF4处理可降低亚铁离子、丙二醛（MDA）和活性氧（ROS）水平，有效抑制PC12细胞中的铁死亡。此外，给予Rm-PF4可减轻ICH小鼠的神经功能障碍、神经元损伤和脑水肿，同时抑制神经元铁死亡。机制上，Rm-PF4激活了CXCR3/PI3K/AKT/Nrf2通路，在ICH小鼠和经血红素处理的PC12细胞中，CXCR3拮抗剂均减弱了这种保护作用。总之，PF4通过激活CXCR3/PI3K/AKT/Nrf2通路减轻小鼠模型和PC12细胞中ICH诱导的神经元铁死亡。
