Exploring the neuroprotective mechanism of lumbrokinase against ischemic stroke based on network pharmacology,...

Ischemic stroke is an acute cerebrovascular disease that remains a major cause of death and long-term disability worldwide. In China and across East Asia, Lumbricus (earthworm) has long been used in traditional medicine for the management of stroke and related cerebrovascular disorders. As a major bioactive component of Lumbricus, lumbrokinase exhibits thrombolytic and anticoagulant activities. The present study investigated whether LK exerts neuroprotective effects in ischemic stroke by modulating TLR4/NF-κB-related inflammatory signaling.

This study aimed to elucidate the neuroprotective mechanism of lumbrokinase against ischemic stroke.

Candidate targets and signaling pathways were predicted by network pharmacology, and binding interactions were assessed by molecular docking. A rat middle cerebral artery occlusion (MCAO) model was then established to evaluate the neuroprotective efficacy of lumbrokinase. Neurological deficits, infarct volume, pathway-related proteins, and inflammatory cytokines were evaluated by behavioral tests, TTC staining, Western blot, and ELISA.

Network analysis identified 49 overlapping inflammation-related targets significantly enriched in TLR4/NF-κB-related pathways. Molecular docking revealed favorable binding affinities (all < -5.0 kcal/mol) between lumbrokinase and core inflammatory mediators, including PTGS2, TRAF6, MyD88, and TLR4. In the MCAO model, lumbrokinase improved neurological function and reduced infarct volume across three doses, with the medium dose (12,000 U/kg) providing the most consistent benefit. Lumbrokinase also decreased the expression of TLR4, MyD88, TRAF6, TAK1 and p-NF-κB/NF-κB, as well as brain levels of TNF-α, IL-1β and IL-6.

Lumbrokinase may confer neuroprotection in ischemic stroke, partly in association with attenuation of TLR4/NF-κB-related neuroinflammation in the ischemic brain.