Objective To explore the mechanism by which Taohong Siwu Decoction (桃红四物汤, THSWD) alleviates cerebral ischemia-reperfusion injury (CIRI) through protecting the integrity and functionality of glycocalyx of brain microvascular endothelial cells. Methods A model of middle cerebral artery occlusion-reperfusion (MCAO/R) was established to simulate CIRI injury. SD rats were randomly divided into sham group, model group, THSWD low-, medium-, high-dose (4.5, 9.0, 18.0 g/kg) groups and nimodipine (20 mg/kg) group, and drugs were given for intervention. After 7 d of modeling, Longa grading system was used for neurological function evaluation, 2,3,5-triphenyltetrazolium chloride (TTC) staining was used to assess cerebral infarction volume. The motion function was evaluated through corner experiments and balance beam experiments. Laser speckle and small animal super-resolution imaging were used to detect cerebral blood flow and cerebral vascular density, Evans blue leak assay combined with Western blotting were to evaluate blood-brain barrier integrity. Glycocalyx morphology was observed using transmission electron microscopy. The expression of glycocalyx-related indicator such as syndecan-1 (SDC-1), hyaluronic acid (HA), heparan sulfate (HS), chondroitin sulfate (CS) and core metabolic enzymes of glycocalyx such as heparanase (HPSE), hyaluronidase 2 (Hyal2), neuraminidase 1 (Neu1), matrix metalloproteinase-9 (MMP-9) were detected by immunofluorescence, immunohistochemistry, Western blotting and ELISA, and the effects of HPSE and MMP-9 were verified by enzyme inhibitors. Results Compared with model group, THSWD significantly reduced the cerebral infarction volume in MCAO/R rats (P < 0.05, 0.001), improved neurological function and limb coordination ability (P < 0.05, 0.01, 0.001), alleviated weight loss, improved cerebral blood flow and microvascular generation in the infarct area, reduced blood-brain barrier leakage (P < 0.01, 0.001), regulated vascular dilation and contraction function (P < 0.01, 0.001). Meanwhile, THSWD could significantly reduce the shedding of core components of glycocalyx (CS, HA, HS, SDC-1) into the serum (P < 0.01, 0.001), and maintain the integrity of glycocalyx by inhibiting the activities of HPSE and MMP-9. Conclusion THSWD reduces the shedding of glycocalyx by inhibiting the activity of HPSE and MMP-9, thereby maintaining the integrity of blood-brain barrier and minimizing stroke damage.

Taohong Siwu Decoction  /  ischemic stroke  /  glycocalyx  /  blood-brain barrier  /  heparanase  /  matrix metalloproteinase-9  /  hydroxysafflor yellow A  /  amygdalin  /  paeoniflorin  /  acteoside  /  ferulic acid  /  ligustilide