Chen Junren, Wu Liujun, Xie Xiaofang, Peng Cheng
State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Front Pharmacol. 2024 Dec 24;15:1504109. doi: 10.3389/fphar.2024.1504109. eCollection 2024.
Stroke is a debilitating disease and the second leading cause of death worldwide, of which ischemic stroke is the dominant type. L., also known as safflower, has been used to treat cerebrovascular diseases, especially ischemic stroke in many Asian countries. However, the underlying mechanisms of safflower in preventing ischemic stroke remains elusive. This study aims to elucidate the potential of safflower as a drug candidate for the prevention of ischemic stroke and to delineate its protective effects and potential mechanisms in a rat model of cerebral ischemia-reperfusion injury (CI/RI).
The aqueous extract of safflower (AESF) was verified using HPLC-UV, HPLC-MS, and TLC. The inhibitory effect of AESF on platelet aggregation was detected and in zebrafish and mice. A CI/RI model in rats was established by middle cerebral artery occlusion and reperfusion to study the protective effect of AESF on ischemic stroke. 2,3,5-triphenyltetrazolium chloride, hematoxylin and eosin, and Nissl's staining were employed to evaluate the pathological changes of brain tissue. In addition, metabolomics, ELISA, and Western blot were used to uncover the molecular alteration induced by AESF.
AESF significantly inhibited platelet aggregation , reduced the thrombogenesis in zebrafish, and prolonged clotting time in mice. In addition, AESF alleviated neurological dysfunction, cerebral oedema, cerebral infarct size, cerebral histopathological damage induced by ischemia-reperfusion, improved neuronal survival, increased serum levels of SOD and CAT, and decreased levels of iNOS and NO. Metabolomics revealed that AESF attenuated the metabolic disturbances in brain caused by I/R injury via regulating 38 metabolites particularly related to the arachidonic acid (AA) metabolism. Moreover, AESF elevated the serum levels of 6-keto-PGF, a pivotal metabolite of AA, downregulated the protein expression of p53, Bax, cleaved caspase-9, cleaved caspase-3, and cleaved caspase-8, and upregulated that of Bcl-2.
AESF mitigated CI/RI through preventing platelet aggregation, alleviating oxidative stress, and suppressing apoptosis partially via modulating AA metabolism/p53-mediated apoptosis axis.
中风是一种使人衰弱的疾病，是全球第二大死因，其中缺血性中风是主要类型。红花在许多亚洲国家已被用于治疗脑血管疾病，尤其是缺血性中风。然而，红花预防缺血性中风的潜在机制仍不清楚。本研究旨在阐明红花作为预防缺血性中风候选药物的潜力，并在大鼠脑缺血再灌注损伤（CI/RI）模型中描述其保护作用和潜在机制。
采用高效液相色谱-紫外检测法（HPLC-UV）、高效液相色谱-质谱联用法（HPLC-MS）和薄层色谱法（TLC）对红花水提取物（AESF）进行验证。检测AESF对斑马鱼和小鼠血小板聚集的抑制作用。通过大脑中动脉闭塞和再灌注建立大鼠CI/RI模型，以研究AESF对缺血性中风的保护作用。采用2,3,5-三苯基氯化四氮唑、苏木精-伊红染色和尼氏染色评估脑组织的病理变化。此外，利用代谢组学、酶联免疫吸附测定法（ELISA）和蛋白质免疫印迹法（Western blot）揭示AESF诱导的分子改变。
AESF显著抑制血小板聚集，减少斑马鱼体内血栓形成，并延长小鼠凝血时间。此外，AESF减轻了缺血再灌注诱导的神经功能障碍、脑水肿、脑梗死体积、脑组织病理损伤，改善了神经元存活，提高了血清超氧化物歧化酶（SOD）和过氧化氢酶（CAT）水平，并降低了诱导型一氧化氮合酶（iNOS）和一氧化氮（NO）水平。代谢组学显示，AESF通过调节38种与花生四烯酸（AA）代谢特别相关的代谢物，减轻了I/R损伤引起的脑内代谢紊乱。此外，AESF提高血清中AA的关键代谢物6-酮-前列腺素F1α（6-keto-PGF）水平，下调p53、Bax、裂解的半胱天冬酶-9、裂解的半胱天冬酶-3和裂解的半胱天冬酶-8的蛋白表达，并上调Bcl-2的蛋白表达。
AESF通过预防血小板聚集、减轻氧化应激以及部分通过调节AA代谢/p53介导的凋亡轴抑制凋亡，从而减轻CI/RI。
