Safety And Efficacy Of Ginkgolide Injection Combined With Alteplase In Thromboembolic Stroke Rats

Background and objective:Currently,the treatment of acute ischemic stroke（AIS）focuses on revascularization and neuroprotection.According to the recommendations of the《Chinese guidelines for diagnosis and treatment of acute ischemic stroke 2018》and the 《European Stroke Organisation（ESO）guidelines on intravenous thrombolysis for acute ischaemic stroke 2021》,recombinant tissue plasminogen activator（rtPA）intravenous thrombolytic therapy is still the most recommended treatment for patients with acute cerebral infarction.However,only about 50% of patients with acute cerebral infarction have a good prognosis,and a large number of patients still do not benefit from rtPA.Therefore,drug combination application to improve the effect of rtPA intravenous thrombolysis is a feasible research direction.Pharmacological studies have shown that ginkgo biloba extract has anti-inflammatory and other neuroprotective effects in various cardiovascular and nervous system diseases.However,further studies are needed to determine whether intravenously Ginkgolide Injection（GI）can improve the prognosis of thrombolysis.In this study,we intend to establish a rat of thromboembolic stroke model（TSM）,investigate the safety and efficacy of GI combined with rtPA in the treatment of TSM rats,and further explore its possible mechanism,in order to improve the prognosis of patients receiving intravenous thrombolysis with rtPA.Methods:Seventy-four male SD rats（300-350 g）were included and randomly divided into Sham group,TSM group,TSM+GI group,TSM+rtPA group,and TSM+rtPA+GI group.The rats in the TSM+GI group were treated with ginkgolide injection for 7 days after successful modeling,the TSM+rtPA group received rtPA thrombolysis after successful modeling,and the TSM+rtPA+GI group received rtPA thrombolysis and ginkgolide injection for 7 days after successful modeling.The neurobehavioral scores of the rats in each group were measured by Zea Longa score at 1,3,and 7 days after the modeling;the hemorrhagic transformation of the rats after thrombolysis was assessed by measuring the hemoglobin level at 24 hours after the modeling and the blood-brain barrier permeability of the rats after thrombolysis was measured by Evan’s blue.The brain infarct volume was measured by TTC staining,the histological changes in the cortex were assessed by HE staining,and the expression of NLRP3 and cleaved caspase-1 protein in each group was measured by Western blot.Results:At 1,3,and 7 days after modeling,the neurobehavioral scores were significantly lower in the TSM+GI group,TSM+rtPA group,and TSM+rtPA+GI group compared with the TSM group（P<0.05）,and at 7 days after modeling,the neurobehavioral scores were significantly lower in the TSM+rtPA+GI group compared with the TSM+rtPA group（P<0.01）;At 24 hours after modeling,there was no significant difference between the TSM+rtPA+GI group and the TSM+rtPA group in terms of hemoglobin volume and Evan’s blue content（P>0.05）;At 7 days after modeling,the cerebral infarct volume in the TSM+rtPA+GI group was significantly lower than that in the TSM+rtPA group（P<0.001）,and the histomorphological improvement of the cerebral cortex in the TSM+rtPA+GI group was more obvious than that in the TSM+rtPA group.NLRP3 and cleaved caspase-1 protein levels were significantly lower in the TSM+rtPA+GI group than in the TSM+rtPA group（P<0.001）.Conclusion:1,Ultra-early GI combined with rtPA treatment in TSM rats is safe;ultra-early GI combined with rtPA treatment significantly improves neurological function and reduces final brain infarct volume in TSM rats.2,GI can improve the prognosis of TSM rats after intravenous thrombolysis by inhibiting NLRP3/cleaved caspase-1 signaling pathway.