ABSTRACT
Objective:To investigate the effect of salvianolate lyophilized injection and Xueshuantong injection (lyophilized) on the permeability of blood-brain barrier (BBB) via inhibition of metallomatrix protease（MMPs） in cerebral ischemia/reperfusion injury rats. Method:The focal cerebral ischemia/reperfusion model in rats was built by middle cerebral artery occlusion/reperfusion (MCAO/R) technique. Male Wistar rats were randomly divided into sham operation group, ischemia/reperfusion (I/R) group, edaravone (Eda, 6 mg·kg-1) group, salvianolate lyophilized injection (SLI, 21 mg·kg-1) group, Xueshuantong (XST, 100 mg·kg-1) group and SLI combined with XST (SLI+XST, 21 mg·kg-1+100 mg·kg-1) group. Drugs were injected via tail vein for 2 d, while sham group and I/R group were injected with the same amount of normal saline. Neurological deficit score, hematoxylin-eosin (HE) staining and Nissl staining were assessed 2 d after MCAO/R. The permeability of BBB was observed by the leakage of IgG/CD31. The expressions of Claudin-5，Occludin，collagen-Ⅳ（Col- Ⅳ），Laminin，Fibronectin were observed by immunofluorescence staining，and MMP-2 and MMP-9 were observed by Western blot. Result:Compared with the I/R group, SLI group, XST group and SLI+XST group showed improvements in neurological deficit score, HE staining and Nissl staining. The leakage of IgG was alleviated; The positive expressions of Claudin-5，Occludin，Col-Ⅳ，Laminin，Fibronectin in ischemic penumbra were significantly up-regulated, while the expressions of MMP-2 and MMP-9 were down-regulated. The effect in improving SLI combined with XST was much better than a single factor. Conclusion:Salvianolate lyophilized injection and Xueshuantong injection (lyophilized) can alleviate cerebral ischemia/reperfusion injury and exert the synergistic effect when they are used in combination. The mechanisms might be associated with the improvement in the permeability of blood-brain barrier by inhibiting MMPs in cerebral ischemia/reperfusion injury rats.
ABSTRACT
Objective: Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion (I/R) injury, resulting in neuronal death. Salvianolate Lyophilized Injection (SLI) and Xueshuantong Injection (Lyophilized) (XST), which have been widely used in the treatment of acutely cerebral infarction clinically in China, exhibit various biological activities. In this study, the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion- reperfusion (MCAO/R) were investigated. Methods: In this study, male Wistar rats were subjected to 1.5 h of middle cerebral artery occlusion followed by reperfusion for 24 h. The rats were randomly divided into the following six groups: normal group (NOR), model group (MOD), SLI group (21 mg/kg, SLI), XST group (100 mg/kg, XST), SLI combined with XST (XST 100 mg/kg + SLI 21 mg/kg, 1X1S), and Edaravone (as a positive control drug, 6 mL/kg, EDI), once a day for 3 d. The neuronal injury, the expression of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (IBA-1), and the changes of pro-inflammatory mediators interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and anti-inflammatory mediator interleukin-10 (IL-10) were observed. Results: 1X1S treatment significantly increased the number of neuron, compared with the MOD group, SLI group and XST group. Gliosis (GFAP and IBA-1) and expression of pro-inflammatory mediators IL-6 and TNF-α were significantly reduced. Meanwhile, 1X1S significantly increased the expression of anti-inflammatory mediator IL-10 in the brains of MCAO/R rats, compared with the MOD group, SLI and XST groups. SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group. Conclusions: This study shows that SLI combined with XST (1X1S) can protect cerebral ischemia-reperfusion injury due to its anti-inflammatory property, and may provide a potential promising new therapeutic strategy for acute ischemic stroke.