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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.
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Objective::To study the degradation of salvianolate lyophilized injection (SLI) and establish a stability-indicating analysis method. Method::UPLC-Q-TOF-MS/MS was used to conduct a qualitative study on the main components of SLI, and a stability-indicating analysis method was established for simultaneous determination of the original components of SLI and its degradation products. The stability of SLI were systematically assessed under physicochemical conditions of high temperature, oxidation, metal ions. Result::Totally 13 main active ingredients in SLI were identified, and a semi-quantitative analysis was performed. Under the conditions of high temperature, oxidation, light, trivalent ion and divalent ion, 6, 4, 3, 4 and 1 new degradation products were added respectively. The established stability-indicating analysis method can simultaneously determine the degradation products of the main components and their active components in SLI, with a good separation effect. Conclusion::According to the degradation mechanism of the main ingredients in SLI, macromolecular polyphenol acid compounds are degraded into small molecular compounds, such as tanshinol and protocatechu aldehyde by a series of reactions, like benzofuran open-loop, hydrolysis of ester bond and removal of DSS. The stability-indicating analysis method can be used for the stability quality control of traditional Chinese medicine Salvianolate Lyophilized Injection (SLI).
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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.
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To investigate the effect of Salvianolate Lyophilized Injection (SLI) combined with Xueshuantong Injection (XST) on expression of astrocytes and microglia in cerebral ischemia-reperfusion injury rats. Methods The Wistar rats (250-300 g, male) were randomly divided into six groups: control group, model group, Edaravone group (6 mg/kg, EDI), SLI group (21 mg/kg), XST group (100 mg/kg), and SLI+XST group (1X1S, 21 mg/kg and 100 mg/kg). Rat model of cerebral ischemia-reperfusion injury was created by the middle cerebral artery occlusion (MCAO) by longa method. Drugs were administered tail intravenous injection once a day for 3 d starting from 24 h after reperfusion. The body weight, neurological deficit scores and survival percentage were observed in 3 d after the cerebral ischemia. The expression of GFAP and IBA-1 was determined at 3 d by immunofluorescence. The complicated compound-target-stroke network of SLI and XST was constructed and analyzed by IPA. Results Compared with the model group, 1X1S could significantly improve the neurological dysfunction, increase the body weight, and inhibit the expression of GFAP and IBA-1 in ischemic penumbra (P < 0.01), IPA reveals the molecular mechanism of SLI and XST in the active ingredient and related targets. Conclusion 1X1S has significant protection on cerebral ischemia reperfusion injury in rats, which may be related to the inhibition of the expression of GFAP and IBA-1 and high mobility group box-1 signaling and Interleukine-8 signaling.
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Aim To investigate the effect of salvianolate syophilized injection on brain tissue gene expression profiles in stroke of diabetic rats.Methods T1DM was induced in adult male Wistar rats by injecting streptozotocin.T1DM rats were then subjected to 90 minutes of middle cerebral artery occlusion(MCAO).The rats were randomly assigned to sham group(DM+Sham),ischemia-reperfusion group(DM+ MCAO/R),edaravone group(6 mg·kg-1,ED)and salvianolate lyophilized injection treatment group(5.25,10.5,21 mg·kg-1,SLI)with 13 rats in each group.Drugs were administered by tail vein injection 3 hours after MCAO/R,daily and lasting for 14 days.Infarct volume and gene expression in the brain tissue were detected by TTC staining and the gene chip technique.Results Compared with DM+Sham group,67 differential expressed genes were detected in the DM+MCAO/R group,among which 41 genes were up-regulated and 26 genes were down-regulated.Compared with DM+MCAO/R group,59 differential expressed genes were detected in the SLI(21 mg·kg-1)group,among which 45 genes were up-regulated and 14 genes were down-regulated.Hierarchical cluster results suggested that a number of genes were significantly changed in T1DM rats,such as Ly6i,Pax7 and Irx2.Effects of SLI on the stroke in T1DM rats were majorly related to coagulation and hemostasis system,inflammatory cytokines,oxidative stress,substance metabolism,angiogenesis and signal transduction.Conclusion Salvianolate lyophilized injection protects against focal cerebral ischemia/reperfusion injury in type 1 diabetic rats through regulation of the coagulation and hemostasis system,inflammatory cytokines,oxidative stress,substance metabolism,angiogenesis and signal transduction.
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Objective To investigate the effects of salvianolate lyophilized injection(SLI) on neural functional recovery and the expression of microtubule associated protein-2(MAP2) after focal cerebral ischemia-reperfusion injury in the diabetic rats.Methods Diabetes model was made by intraperitoneal injection of streptozotocin (STZ) and cerebral ischemia-reperfusion model was developed by Longa suture occluded method in the middle cerebral artery of diabetic rats.The rats were randomly divided into five groups: sham operation group, model group, SLI (21.0 mg.kg-1,10.5 mg.kg-1) treatment groups, and edaravone (6 mg.kg-1) treatment group.3 hours after ischemia,rats were respectively given normal saline or drugs followed by the injection once a day for 14 days and the neurological impairment was assessed.2 h after the last injection,the rats were decapitated and the brains were collected.The expression of MAP2 protein and mRNA in the bilateral hippocampal ischemia and infarcted area was detected with immunohistochemistry and RT-PCR.Results Severe neurological dysfunction was found in diabetic rats that had been subjected to cerebral ischemic injury (1.850±0.457).A significant improvement on neurological function was found in the SLI treatment groups (1.581 ± 0.314, 1.345 ± 0.425) compared with model group(P＜0.01, P＜0.05).Moreover,the expression of MAP2 in ischemia bilateral hippocampal CA1 and penumbra was represented by the average optical density value respectively (0.743±0.250,0.561± 0.224).In the hippocampal CA1 region, the number of MAP2-positive cells (0.781 ± 0.420 , 0.851 ± 0.136) in the treatment group showed significant increase than those in model group (P＜0.01, P＜0.05).In the ischemic penumbra region,the number of MAP2-positive cells (0.753±0.235,1.203±0.326) in the treatment group showed significant increase than those in model group (P＜0.01, P＜0.05).Conclusion The SLI can promote the post-injury neurocognitive function in diabetic rats.The increase of MAP2 expression may be involved in the mechanisms.