Neuroprotective Effect And Its Mechanism Of Cattle Encephalon Glycoside And Ignotin In Intracerebral Hemorrhage Rats

BackgroundThe morbidity,mortality,and disability associated with intraventricular hemorrhage(IVH)secondary to intracerebral hemorrhage(ICH)represent a global burden.To date,there is no effective therapy for ICH other than supportive care.The results of several clinical trials for cerebral hemorrhage surgery and blood pressure management were negative results.Furthermore,neuroprotective therapies in ICH from bench to bedside were declared failure.At present,basic research also focused on pure cerebral hemorrhage,for cerebral hemorrhage into the ventricle of this serious cerebral hemorrhage subtype of the study is still a blank.Facing the complex pathological damage mechanism after cerebral hemorrhage,the current single target,single molecular pathway of neuroprotective drug research is difficult to be effective.Cattle encephalon glycoside and ignotin(CEGI)is a complex multi-target neuroprotective drug with a high degree of blood-brain barrier permeability.Its main components are polypeptides,various gangliosides,free amino acids,hypoxanthine and so on.CEGI is used in clinical treatment of central nervous system and peripheral nervous system diseases.Basic research has also confirmed that it has neuroprotective effects in various animal models such as Alzheimer's disease,traumatic brain injury,and retinal degeneration,but its neuroprotective effects and mechanisms in hemorrhagic stroke are still unclear.In the first part of this study,we established a standardized model of IVH/ICH and study the neuroprotective effects of different doses of CEGI on the recovery of neurological function after intracerebral hemorrhage in rats.In the second part,we study the possible mechanism and potential target of neuroprotective effect of CEGI in a rat model of IVH/ICH,and provide a new method for the clinical treatment of IVH/ICH.Part ? Established the rat model of IVH/ICH by Type IV collagenase and explored the optimal therapeutic dose of CEGIObjective Through the improvement of traditional rat intracerebral hemorrhage model,we established the rat standardized model of IVH/ICH.And we evaluated the neuroprotective effect of different doses of multi-target compound neuroprotective drugs on the recovery of neurological function after IVH/ICH in rats by behavior assessments.Methods Based on method proposed by Rosenberg et al and our previous work,we adjusted the injection coordinates and 0.2U type IV collagenase was infused into the right basal ganglia(coordinates: 0.2 mm posterior,3.0 mm lateral,and 6.0 mm depth to the bregma).3 days after that,we observe whether the hematoma to break into the ventricle by general specimen.The rats were randomly divided into five treatment groups: sham,ICH + vehicle(4 ml/kg/d),ICH + CEGI(1 ml/ kg/d),ICH + CEGI(4 ml/kg/d),and ICH + GM-1(50 g/kg/d).Neurological deficits were evaluated on days 3,7,and 14 post-ICH induction using an 18-point score system named the modified Garcia Scale and a percentage system named the Corner turn test.Results 1.3 days after surgery,hematoma formed and broke into the lateral ventricle with ventricular dilatation;2.Ganglioside and 4ml / kg CEGI can significantly improve the prognosis of IVH/ICH in rats;3.There was no significant difference in the neurological deficit between the ICH + CEGI(1 ml/ kg/d)group and the ICH + vehicle(4 ml/kg/d)group.Conclusion The effect of the method which established the rat model of IVH/ICH was exact.And successfully simulated the clinical IVH/ICH which is the most severe hemorrhagic stroke subtype.Part ? Study on the mechanism of CEGI in the treatment of IVH/ICH in rat Objective In the first part of the study,we found that the neurological deficits of IVH/ICH in rat were significantly improved by 4ml/kg CEGI intraperitoneal injection for 14 days.Recent literatures indicate that several major constituents of the CEGI have antioxidant effects,axonal regeneration,and myelination.Therefore,this study intends to explore the possible mechanism of CEGI in the treatment of IVH/ICH in rat.Methods Rats were randomly assigned into three treatment groups: sham,ICH + vehicle(4 ml/kg/d)and ICH + CEGI(4 ml/kg/d).The first administration was given intraperitoneally 2 hours after surgery and for 14 days.The changes of hematoma and lateral ventricle volume were observed by microsatellite 7.0T nuclear magnetic resonance imaging at 3 days,7 days and 14 days after operation.At 7 days and 14 days after operation,laser confocal microscopy and transmission electron microscopy were used to observe the white matter injury around hematoma and at cortex.Results 1.The absorption rate of the hematoma was significantly higher in the ICH + CEGI 4ml/kg group than in the ICH group at day 14.2.At day 14,the volume of the lateral ventricles in the ICH + CEGI 4 ml/kg group was significantly smaller compared with that in the ICH group.The long-term occurrence of severe hydrocephalus was significantly decreased in the ICH + CEGI 4 ml/kg group compared with the ICH group.3.The expression of myelin basic protein in the cortex was significantly increased at 7 and 14 days after operation,and the ultrastructural damage of the white matter fibers around the hematoma was improved.Conclusion CEGI treatment significantly ameliorated white matter fiber damage,promoted hematoma absorption,reduced the lateral ventricular enlargement,reduce d the incidence of long-term severe hydrocephalus and alleviated the neurobehavioral dysfunction post-ICH induction.Our results demonstrate that CEGI has significant neuroprotective effects in a rat model of IVH/ICH.Therefore,it can be used as a candidate drug for the clinical treatment of IVH/ICH.