Department of Neurosurgery, Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan, China.
J Biochem Mol Toxicol. 2023 May;37(5):e23317. doi: 10.1002/jbt.23317. Epub 2023 Mar 5.
Early brain injury (EBI) is associated with the adverse prognosis of subarachnoid hemorrhage (SAH) patients. The key bioactive component of the Chinese herbal medicine Artemisia asiatica Nakai (Asteraceae) is eupatilin. Recent research reports that eupatilin suppresses inflammatory responses induced by intracranial hemorrhage. This work is performed to validate whether eupatilin can attenuate EBI and deciphers its mechanism. A SAH rat model was established by intravascular perforation in vivo. At 6 h after SAH in rats, 10 mg/kg eupatilin was injected into the rats via the caudal vein. A Sham group was set as the control. In vitro, BV2 microglia was treated with 10 μM Oxyhemoglobin (OxyHb) for 24 h, followed by 50 μM eupatilin treatment for 24 h. The SAH grade, brain water content, neurological score, and blood-brain barrier (BBB) permeability of the rats were measured 24 h later. The content of proinflammatory factors was detected via enzyme-linked immunosorbent assay. Western blot analysis was conducted to analyze the expression levels of TLR4/MyD88/NF-κB pathway-associated proteins. In vivo, eupatilin administration alleviated neurological injury, and decreased brain edema and BBB injury after SAH in rats. Eupatilin markedly reduced the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), and suppressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in the SAH rats' cerebral tissues. Eupatilin treatment also reduced the levels of IL-1β, IL-6, and TNF-α, and repressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in OxyHb-induced BV2 microglia. Additionally, pyrrolidine dithiocarbamate or resatorvid enhanced the suppressive effects of eupatilin on OxyHb-induced inflammatory responses in BV2 microglia. Eupatilin ameliorates SAH-induced EBI via modulating the TLR4/MyD88/NF-κB pathway in rat model.
早期脑损伤（EBI）与蛛网膜下腔出血（SAH）患者的不良预后相关。中草药茵陈蒿（菊科）的关键生物活性成分是芹菜素。最近的研究报告表明，芹菜素可抑制颅内出血引起的炎症反应。本工作旨在验证芹菜素是否能减轻 EBI，并解析其机制。通过体内血管穿孔建立 SAH 大鼠模型。在大鼠 SAH 后 6 小时，通过尾静脉向大鼠注射 10mg/kg 的芹菜素。假手术组作为对照。在体外，用 10μM 氧合血红蛋白（OxyHb）处理 BV2 小胶质细胞 24 小时，然后用 50μM 芹菜素处理 24 小时。24 小时后测量大鼠的 SAH 分级、脑水含量、神经评分和血脑屏障（BBB）通透性。通过酶联免疫吸附试验检测促炎因子的含量。通过 Western blot 分析检测 TLR4/MyD88/NF-κB 通路相关蛋白的表达水平。体内，芹菜素给药可减轻 SAH 后大鼠的神经损伤，降低脑水肿和 BBB 损伤。芹菜素显著降低 SAH 大鼠脑组织中白细胞介素-1β（IL-1β）、IL-6 和肿瘤坏死因子-α（TNF-α）的水平，并抑制 SAH 大鼠脑组织中 MyD88、TLR4 和 p-NF-κB p65 的表达水平。芹菜素处理还降低了 OxyHb 诱导的 BV2 小胶质细胞中 IL-1β、IL-6 和 TNF-α的水平，并抑制了 OxyHb 诱导的 BV2 小胶质细胞中 MyD88、TLR4 和 p-NF-κB p65 的表达水平。此外，吡咯烷二硫代氨基甲酸盐或 resatorvid 增强了芹菜素对 OxyHb 诱导的 BV2 小胶质细胞炎症反应的抑制作用。芹菜素通过调节 TLR4/MyD88/NF-κB 通路改善 SAH 诱导的 EBI。
