Ginsenoside Rh2-modified liposomes for targeted delivery of Puerarin alleviate brain ischemia-reperfusion injury

Brain ischemia poses a significant unmet medical need, demanding novel therapeutic approaches. Puerarin (Pue), despite its potential for treating brain disorders, suffers from poor blood-brain barrier (BBB) permeability due to its low oil/water partition coefficient. To overcome this, we developed a novel ginsenoside Rh2-based liposome formulation (Rh2-Pue-LP) to enhance Pue delivery to the ischemic brain. A rat model of middle cerebral artery occlusion-reperfusion (MCAO/R) was established. Neurological deficits were evaluated using the Longa scoring system 24 h post-MCAO. After seven days of tail-vein administration of Rh2-Pue-LP, the following analyses were performed: TTC staining to assess cerebral infarct volume, HE and TUNEL staining to examine hippocampal histopathology, ELISA to quantify serum levels of IL-1β, TNF-α, and IL-6, and an LDH assay to measure serum lactate dehydrogenase activity reflecting cellular injury and pyroptosis, immunofluorescence to detect NLRP3 expression, and immunohistochemistry to evaluate the activation of JAK2-STAT3 and expression of inflammatory cytokines. Additionally, this study was conducted to further verify the targeting ability and safety of the formulation. Our results showed Rh2-Pue-LP treatment reduced infarct volume, improved neurological function, and decreased serum levels of inflammatory cytokines (IL-1β, TNF-α, IL-6), and markedly reduced serum LDH release Histological examination revealed better-preserved hippocampal neurons. Rh2-Pue-LP inhibited the JAK2-STAT3 signaling pathway and NLRP3 inflammasome expression, suppressing microglial activation and neuronal apoptosis. Additionally, Rh2-Pue-LP exhibited stronger brain targeting ability with no significant biotoxicity in vivo. Rh2-Pue-LP represents a promising strategy for treating ischemic stroke by enhancing Pue delivery and exerting potent neuroprotective effects.