中风醒脑液通过抑制神经元铁死亡发挥抗脑出血损伤的作用机制研究  

Zhongfeng Xingnao Decoction alleviates intracerebral haemorrhage-induced brain injury via inhibiting neuronal ferroptosis

作　　者：李孟英 郭建文 邵翀羽 泮智勇 陈天杭 周科峰 周惠芬 万海同[1,3,4] LI Meng-ying;GUO Jian-wen;SHAO Chong-yu;PAN Zhi-yong;CHEN Tian-hang;ZHOU Ke-feng;ZHOU Hui-fen;WAN Hai-tong(the First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China;School of Life Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Provincial Key Laboratory of Traditional Chinese Medicine Cardiovascular and Cerebrovascular Diseases,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)

机构地区：[1]浙江中医药大学第一附属医院,浙江杭州310006 [2]浙江中医药大学生命科学学院,浙江杭州310053 [3]浙江中医药大学基础医学院,浙江杭州310053 [4]全省中医心脑血管病重点实验室,浙江杭州310053 [5]广州中医药大学第二附属医院省部共建中医湿证国家重点实验室,广东广州510120 [6]广州中医药大学第二附属医院脑病中心,广东广州510120

出　　处：《中国中药杂志》2024年第17期4723-4733,共11页China Journal of Chinese Materia Medica

基　　金：广东省重点领域研发计划项目(2020B1111100009);国家自然科学基金项目(81974559);广东省自然科学基金项目(2020A1515010992)。

摘　　要：中风醒脑液在临床上已被用于治疗脑出血。然而,目前对中风醒脑液的药理作用尚不明确。探讨中风醒脑液的药理作用对指导脑血管疾病的治疗至关重要。向大鼠右侧尾状核注射Ⅶ型胶原酶建立体内脑出血模型。将SD大鼠随机分成5组,用Bederson评分法评估大鼠术后的神经功能缺损情况。核磁共振成像(MRI)用于评估脑出血的体积。苏木精-伊红(HE)染色观察脑组织病理改变,透射电镜(TEM)观察脑组织超微结构变化。流式细胞术测定活性氧(ROS)水平。免疫荧光双染法检测脑组织血肿周围神经元中铁转运蛋白标记物谷胱甘肽过氧化物酶4(GPX4)的表达。Western blot法检测脑出血后RNA解旋酶和ATP酶1(UPF1)、铁泵蛋白(FPN)、酰基辅酶A连接酶4(ACSL4)、环氧化酶2(COX2)、GPX4、NADPH氧化酶1(NOX1)和胱氨酸/谷氨酸逆向转运蛋白(SLC7A11)的表达。与模型组相比,中风醒脑液治疗5 d后大鼠神经功能障碍明显减轻,血肿体积减小,脑出血病理损伤减轻;与此同时,Fe^(2+)和氧化应激水平显著降低,抑制铁死亡的蛋白表达量显著增加。综上,中风醒脑液可通过显著抑制神经元铁死亡改善大鼠脑出血的预后,为中风醒脑液的临床应用提供了有价值的指导。中风醒脑液的抗铁死亡作用可能与促进膜转运蛋白SLC7A11和FPN的表达有关。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 ofⅦcollagenase 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),cyclooxygenase-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 Fe2+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.

关 键 词：脑出血 铁死亡 中风醒脑液 神经元死亡 

分 类 号：R285[医药卫生—中药学]