Calycosin-7-O-β-D-Glucoside Facilitates Axonal Regrowth and Functional Recovery via Rho/ROCK Pathway Inhibition After...

Calycosin-7-O-β-D-glucoside (CG), a bioactive compound extracted from the traditional Chinese herb Astragalus (AR), exhibits diverse biological activities, including anti-oxidative and anti-inflammatory effects, and has shown protective properties in ischemia-reperfusion (I/R) injury. While previous studies have demonstrated that CG mitigates I/R injury primarily through its anti-oxidative and anti-inflammatory actions, its potential role in promoting neuroregeneration-a critical process for stroke recovery-remains unclear, and the underlying mechanisms have yet to be elucidated. In this study, an ischemic stroke model was established in rats via middle cerebral artery occlusion (MCAO). Seven days after CG treatment, cerebral infarct volume was assessed using triphenyltetrazolium chloride (TTC) staining, while neurological function was evaluated through behavioral tests. Nissl staining and Bielschowsky silver staining were employed to examine neuronal damage and axonal loss, and immunofluorescence was used to assess axonal regeneration. The expression of key proteins in the Rho/ROCK signaling pathway was analyzed by Western blotting (WB) and quantitative real-time PCR (qRT-PCR). CG treatment significantly reduced infarct volume, promoted axonal regeneration, improved neurological outcomes, and modulated the expression of RGMa, Rho, ROCK, and CRMP2. Collectively, these findings provide the first evidence that CG facilitates axonal regeneration and neurological recovery after cerebral ischemia, at least in part by inhibiting activation of the Rho/ROCK pathway, highlighting its potential as a therapeutic agent for ischemic stroke.