Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu, 610041, China.
Department of Cerebrovascular Disease, The People's Hospital of Leshan, Leshan, 614000, China.
Curr Neurovasc Res. 2024;21(1):74-85. doi: 10.2174/0115672026295640240212095049.
Intracerebral hemorrhage (ICH) is one of the most common subtypes of stroke.
This study aimed to investigate the mechanism of Astragaloside IV (AS-IV) on inflammatory injury after ICH.
The ICH model was established by the injection of collagenase and treated with ASIV (20 mg/kg or 40 mg/kg). The neurological function, water content of the bilateral cerebral hemisphere and cerebellum, and pathological changes in brain tissue were assessed. The levels of interleukin-1 beta (IL-1β), IL-18, tumor necrosis factor-alpha, interferon-gamma, and IL-10 were detected by enzyme-linked immunosorbent assay. The levels of Kruppel-like factor 2 (KLF2), NOD-like receptor family pyrin domain containing 3 (NLRP3), GSDMD-N, and cleaved-caspase-1 were detected by reverse transcription-quantitative polymerase chain reaction and Western blot assay. The binding relationship between KLF2 and NLRP3 was verified by chromatin-immunoprecipitation and dual-luciferase assays. KLF2 inhibition or NLRP3 overexpression was achieved in mice to observe pathological changes.
The decreased neurological function, increased water content, severe pathological damage, and inflammatory response were observed in mice after ICH, with increased levels of NLRP3/GSDMD-N/cleaved-caspase-1/IL-1β/IL-18 and poorly-expressed KLF2 in brain tissue. After AS-IV treatment, the neurological dysfunction, high brain water content, inflammatory response, and pyroptosis were alleviated, while KLF2 expression was increased. KLF2 bonded to the NLRP3 promoter region and inhibited its transcription. Down-regulation of KLF2 or upregulation of NLRP3 reversed the effect of AS-IV on inhibiting pyroptosis and reducing inflammatory injury in mice after ICH.
AS-IV inhibited NLRP3-mediated pyroptosis by promoting KLF2 expression and alleviated inflammatory injury in mice after ICH.
脑出血(ICH)是中风最常见的亚型之一。
本研究旨在探讨黄芪甲苷(AS-IV)对脑出血后炎症损伤的作用机制。
采用胶原酶注射法建立 ICH 模型,并给予 AS-IV(20mg/kg 或 40mg/kg)治疗。评估神经功能、双侧大脑半球和小脑含水量以及脑组织病理变化。通过酶联免疫吸附试验检测白细胞介素-1β(IL-1β)、IL-18、肿瘤坏死因子-α、干扰素-γ和 IL-10 水平。通过逆转录定量聚合酶链反应和 Western blot 检测 Kruppel 样因子 2(KLF2)、NOD 样受体家族含 pyrin 结构域蛋白 3(NLRP3)、Gasdermin D-N(GSDMD-N)和切割-caspase-1 的水平。通过染色质免疫沉淀和双荧光素酶报告基因实验验证 KLF2 与 NLRP3 之间的结合关系。通过在小鼠中抑制 KLF2 或过表达 NLRP3 观察病理变化。
ICH 后小鼠神经功能下降,脑含水量增加,病理损伤严重,炎症反应增强,脑组织中 NLRP3/GSDMD-N/cleaved-caspase-1/IL-1β/IL-18 水平升高,KLF2 表达降低。AS-IV 治疗后,神经功能障碍、脑含水量高、炎症反应和细胞焦亡减轻,KLF2 表达增加。KLF2 与 NLRP3 启动子区域结合并抑制其转录。下调 KLF2 或上调 NLRP3 逆转了 AS-IV 对抑制小鼠 ICH 后细胞焦亡和减轻炎症损伤的作用。
AS-IV 通过促进 KLF2 表达抑制 NLRP3 介导的细胞焦亡,减轻 ICH 后小鼠的炎症损伤。