Thrombus-Targeted and Reactive Oxygen Species-Responsive Apigenin Liposomes for Ischemic Stroke

Abstract Ischemic stroke remains a leading cause of mortality and long-term disability, while current therapies inadequately address oxidative stress, mitochondrial dysfunction, and blood–brain barrier (BBB) disruption during ischemia and reperfusion. Here, we developed DPCT@LIP-AP, a reactive oxygen species (ROS) responsive liposomal formulation of apigenin incorporating cysteine-arginine-glutamate-lysine-alanine (CREKA) and a ROS-cleavable thioketal linker. DPCT@LIP-AP exhibited favorable colloidal stability, ROS-dependent drug release, enhanced binding to fibrin clots, and preferential accumulation in ischemic brain tissue. In HT22 cells subjected to oxygen-glucose deprivation and reoxygenation (OGD/R), DPCT@LIP-AP reduced intracellular ROS accumulation, preserved mitochondrial membrane potential, restored adenosine triphosphate production, and suppressed the expression of IL-1β, IL-6, and TNF-α. These effects were accompanied by increased Nrf2 and HO-1 expression. In a mouse model of middle cerebral artery occlusion and reperfusion (MCAO/R), intravenous DPCT@LIP-AP reduced infarct volume and brain edema, preserved BBB and neuronal integrity, attenuated neuronal apoptosis, maintained cerebral microvascular structures, and improved neurological and behavioral outcomes. Competitive blockade with free CREKA reduced cerebral accumulation, supporting a CREKA-dependent contribution to lesion-directed delivery. Collectively, the integration of CREKA-mediated fibrin recognition with ROS-responsive apigenin release provides a coordinated strategy for protecting the neurovascular unit after cerebral ischemia and reperfusion. These findings support the further development of DPCT@LIP-AP as a nanotherapeutic platform for ischemic stroke.