Li Meng-Ying, Guo Jian-Wen, Shao Chong-Yu, Pan Zhi-Yong, Chen Tian-Hang, Zhou Ke-Feng, Zhou Hui-Fen, Wan Hai-Tong
School of Life Sciences, Zhejiang Chinese Medical University Hangzhou 310053, China.
State Key Laboratory of Dampness Syndrome of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine Guangzhou 510120, China Department of Neurology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine Guangzhou 510120, China.
Zhongguo Zhong Yao Za Zhi. 2024 Sep;49(17):4723-4733. doi: 10.19540/j.cnki.cjcmm.20240521.401.
Zhongfeng Xingnao Decoction(ZFXN) has been utilized for treating intracerebral hemorrhage(ICH) in China, while the pharmacological mechanism of ZFXN remains unclear. Exploring the pharmacological roles of ZFXN is critical for guiding the treatment of cerebrovascular diseases. In this study, a rat model of ICH was constructed by injection of Ⅶ collagenase in the right caudate nucleus. SD rats were randomly assigned into five groups, and the neurological function of rats was evaluated based on the Bederson score. Magnetic resonance imaging(MRI) was used to assess the volume of ICH. Hematoxylin-eosin(HE) staining and transmission electron microscopy(TEM) were employed to observe the pathological and ultrastructural changes in the brain tissue. The levels of reactive oxygen species(ROS) were measured by flow cytometry. The immunofluorescence assay was employed to detect the expression of glutathione peroxidase 4(GPX4) in neurons surrounding the hematoma. Finally, Western blot was employed to determine the expression of ferroptosis-related proteins upstream frameshift 1(UPF1), ferroportin(FPN), acyl-CoA ligase 4(ACSL4), cyclooxyge-nase-2(COX-2), GPX4, NADPH oxidase 1(NOX1), and solute carrier family 7 member 11(SLC7A11) after ICH. Compared with the model(ICH) group, ZFXN treatment for 5 days attenuated neurological dysfunction, reduced the hematoma volume, and alleviated the pathological changes induced by ICH. Meanwhile, ZFXN lowered the levels of Fe~(2+) and oxidative stress and up-regulated the expression of proteins inhibiting ferroptosis. ZFXN improved the prognosis of ICH in rats by inhibiting neuronal ferroptosis, which provided a valuable guide for the clinical application of ZFXN. ZFXN may inhibit ferroptosis by promoting the expression of SLC7A11 and FPN.
中风醒脑汤（ZFXN）在中国一直用于治疗脑出血（ICH），但其药理机制尚不清楚。探索ZFXN的药理作用对于指导脑血管疾病的治疗至关重要。在本研究中，通过向右侧尾状核注射Ⅶ型胶原酶构建ICH大鼠模型。将SD大鼠随机分为五组，并根据贝德森评分评估大鼠的神经功能。采用磁共振成像（MRI）评估ICH体积。采用苏木精-伊红（HE）染色和透射电子显微镜（TEM）观察脑组织的病理和超微结构变化。通过流式细胞术检测活性氧（ROS）水平。采用免疫荧光法检测血肿周围神经元中谷胱甘肽过氧化物酶4（GPX4）的表达。最后，采用蛋白质印迹法检测脑出血后铁死亡相关蛋白上游移码1（UPF1）、铁转运蛋白（FPN）、酰基辅酶A连接酶4（ACSL4）、环氧化酶-2（COX-2）、GPX4、NADPH氧化酶1（NOX1）和溶质载体家族7成员11（SLC7A11）的表达。与模型（ICH）组相比，ZFXN治疗5天可减轻神经功能障碍，减少血肿体积，并减轻ICH诱导的病理变化。同时，ZFXN降低了Fe2+水平和氧化应激，并上调了抑制铁死亡的蛋白表达。ZFXN通过抑制神经元铁死亡改善了大鼠ICH的预后，为ZFXN的临床应用提供了有价值的指导。ZFXN可能通过促进SLC7A11和FPN的表达来抑制铁死亡。
