Eupatilin alleviates inflammatory response after subarachnoid hemorrhage by inhibition of TLR4/MyD88/NF-κB axis

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 mu M Oxyhemoglobin (OxyHb) for 24 h, followed by 50 mu M 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-kappa B 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-1 beta (IL-1 beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha), and suppressed the expression levels of MyD88, TLR4, and p-NF-kappa B p65 in the SAH rats' cerebral tissues. Eupatilin treatment also reduced the levels of IL-1 beta, IL-6, and TNF-alpha, and repressed the expression levels of MyD88, TLR4, and p-NF-kappa B 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-kappa B pathway in rat model.

Automated summary

This study aims to validate whether eupatilin, the key bioactive component of Artemisia asiatica Nakai, can attenuate early brain injury (EBI) caused by subarachnoid hemorrhage (SAH) and decipher its mechanism. The results showed that eupatilin can alleviate neurological injury, reduce brain edema and blood-brain barrier (BBB) injury, and suppress inflammatory responses. It was also found that eupatilin improves SAH-induced EBI through modulating the TLR4/MyD88/NF-kappa B pathway.