Discovery And Preliminary Mechanism Study Of An Innovative Drug For Treating Ischemic Stroke

Ischemic stroke refers to the lack of cerebral blood supply due to various reasons,resulting in the death of neurons and glial cells,causing ischemic and hypoxic necrosis in local brain tissue,and then causing permanent infarction brain tissue.Ischemic stroke is characterized by a high rate of morbidity,recurrence,disability and mortality.The medical field has been focusing on the research of novel anti-acute ischemic stroke drugs,but the results are not satisfactory.HOPA is a new type of drug independently synthesized by our research group,which has the function of regulating blood oxygen and provides an effective lead compound for the treatment of ischemic stroke.In this study,the intraluminal suture middle cerebral artery occlusion（MCAO）model was selected to establish a mouse cerebral ischemia model and related research was carried out.This model can achieve ischemia-reperfusion and control the time of thrombectomy,which is a good simulation of clinical disease,ischemic stroke.The therapeutic effect of HOPA,a novel blood oxygen regulator,was evaluated in an ischemia-reperfusion model,and the results showed that HOPA treatment can improve neurological deficits and reduce infarct volume significantly.In the behavioral experimental,the motor coordination ability and mortality of the model mice was significantly improved,and neurological impairment,as well as weight loss of the model mice were significantly inhibited by continuous administration of HOPA for 7days.Thereafter,a series of molecular biology experiments were conducted to investigate the mechanism of HOPA preliminarily in the treatment of ischemic stroke.The expression levels of pro-inflammatory cytokines（TNF-α,IL-1β,IL-6,i NOS and COX2）and hypoxia inducible factor-1α（HIF-1α）and the activation of microglia and astrocytes were detected.It is found that HOPA treatment significantly inhibited the expression levels of pro-inflammatory cytokines and HIF-1α and reduced the activation of microglia and astrocytes.In addition,2,5-dihydroxybenzoic acid（DHB）,the possible metabolite of HOPA,was selected to perform the same behavioral and molecular biology experiments to test the mode of action of the HOPA compound in the treatment of ischemic stroke.The results show that DHB has a certain therapeutic effect to improve motor coordination ability and neurological damage in the model mice,but the therapeutic effect is not more significant than that of HOPA.To a certain extent,HOPA is indeed a directly acting component,rather than its metabolite.In summary,the middle cerebral artery occlusion model established by the monofilament method were used as ischemic stroke models in vivo.The behavioral experiments,including neurobehavioral injury score and fatigue rotating bar experiment,show that HOPA can significantly improve the behavioral ability of the damaged mice,block the progress of injury,and achieve certain therapeutic effects.Meanwhile,based on the inflammatory response occurring in ischemic stroke,a preliminary study was conducted to investigate the mechanism for treatment of ischemic stroke with HOPA by detecting the changes of astrocytes,microglia and some pro-inflammatory factors in brain tissue through molecular biology experiments.The results suggest that HOPA have potential clinical application value in the treatment of ischemic stroke and provide a new strategy for the design and development of novel drugs for the treatment of ischemic stroke.