Impression And Effectiveness Of Hephaestin After Middle Cerebral Artery Ischemis-reperfusion In Rats

PartⅠThe establishment of the middle cerebral artery occlusionmodel modelNowadays,the cerebral vascular disease is becoming the third most common death in our country. The disease is a severe disease with high morta-lity and disability,and the low cure rate. Eespecially,alarge area, basal nuclei cerebral infarction has caused very serious infulence to the work and life of the patients.So it is very important to prevention and cure these disease, it can help to early treatment and convalescence in the clinical.So the reliable and reroducible animal models of cerebral ischemia is seriously needed,because it can simulate clinical pathophysiology process.In rencent years,the focal ischemic model is frequently used, it can simulate the process of clinical pathophysiology,thereby it can get the purpose of scientific research,and middle cerebral artery occlusion is the most application in the experimentation. And the model was modified by method of Zealonga.In the experiment,the rabbit beards which was dealt with by polyvinyl alcohol was used to make MCAO. The polyvinyl alcohol was inflation when meet the rat's blood,so it could fit different rat with different body weight and different thickness blood vessel,and the blood flow was blocked more completely. Simultaneously,it can elevate the achievement ratio of the model. The common carotid artery embolism method and the external carotid artery embolism method was used,and through the nerve muscle score, vestibular function score, cerebral infarction volum,to compare the superiority of the two method.Objectiveto make focal cerebral ischumic model of middle cerebral artery in Wistar rats by two methods, and to compare the methods of occlusion of common carotid artery and external carotid artery. And to offer the theory gist for diagnosis and treatment of the ischemic cerebrovascular diseaseMethodHere to selective 70 clearing Wistar rat, and the embolism reperfusion model was divided randomly into 4 groups, control group(5), sham operation group(5), common carotid artery group(30), external carotid artery group(30), and the last two groups were divided randomly into 6 subsections, every subsections has 5 rats,that is 4h,8h,16h,32h,64h,128h after ischemic reper-fusion.The ischemic refusion model was established.The middle artery ischemia model of Wistar rats were made by the modified method of Zealonga.In the experiment,the rabbit beards which was dealt with by polyvinyl alcohol was used to make MCAO via the methods of common carotid artery and external carotid artery. The reperfusion was done after 90min of ischemic.The nerve muscle score method and the vestibule motor function score method was used to score the rat's motor function after 4hour,8hour,16hour,32hour,64hour and 128hour of reperfusion.After 3days,the rats were put to death,and the infarction volume was deterined and compared.ResultThe nerve muscle scorings in common carotid artery group and external carotid artery were 4.1±0.645 and 5.1±0.546, respectively. The scorings between the two methods was significantly different (p<0.05).The vestibule motor function scorings of the two methods were 2.6±0.576 and 3.6±0.519 respectively, The scorings between the two methods was significantly different too, (P<0.05). The infarction volume of the two methods was 22.9%±4.6% and 30.5%±3.5%, and also there was significantly different in the infarction volume between the two groups, (p<0.05).ConclusionBoth the common carotid artery group and external carotid artery were appeared nerve functional impairment and ischemic infarction of brain tissue. Compared with the common carotid artery occlusion group, the neurological behavior scoring of external carotid artery group was higher, the infarction volume was larger, and the stability was better. PARTⅡImpression and effectiveness of Hephaestin after middle cerebral artery ischemis-reperfusion in ratsIron is the sine qua non trace meter in all life,also it is the most affluent transition metal in the human body.The original study discovers that the importance of function of iron that people to know more than people can imagine. There is iron in the every parts of brain,but the distribution is not uniformity.Iron reside in many cell of nervous system,such as neurone, microglia, oligodendrocyte and part of astrocyte,and the oligodendrocyte has the more iron than any other part of brain.The iron in the brain to rise to get up very important physiological effectiveness,it is electron transporter and the important matter of mitochondrion oxidizing reaction and oxygen transport.It participate the synthesize of DNA,RNA, protein and myelin,and it also participate the development and genesis of nerve myelin sheath. Meanwhile,it is indispensable in the synthesis of neurotransmitter,such as dopamine and so on.It is very important to maintain the homeostasis of organism.More and more study has discovered that the homeostasis disequilibrium after acute cerebral ischemia.The redundant redundant iron can mediate a series of patho- physio logy course,and can induce various kinds of disease.Generally,the absorption of iron in the brain tissue via the classic transferrin and transferrin receptor. But recentlt's study shows that the mechanism to take in iron in brain is more complicated than periphery,and the equilibrium mechanism of iron in brain is more complicated than out brain.Besides the main path of transferrin that is mediated by transferrin receptor,the new discovered iron metabolism proteinum such as DMT 1 and Fpl also participate iron metabolism in brain.Hephaestin is a kind of new iron conveying associated protein,it was discovered when studying sex-linked anemic mouse.The expression of hephaestin has conspicuous tissue specificity, the expression in intestinum is height.The experimental result indicated that the expression of Hephaestin mRNA and proteinum were abundance in cerebral cortex, hippocampus, choroid plexus, corpora striata and substantia nigra of rat and mouse.And the latest report say that the people's Hp also expresses in the nervous system of intestinal tract and theβcells of abdominal salivary gland. Hephaestin and Ceruloplasmin are in the same family,which is a kind of ferrous oxidase of cuprine, it can transform the noxious bivalence iron into atoxigenic Ferric.By far,it has reported the changes of expression of Hephaestin after cerebral ischemia of rat.The objective of this study is to observe diversity the diversity law of Hephaestin in different rat and in different time of ischemia reperfusion,and to approach the role in tissue of acute cerebral ischemia and hypoxia.ObjectiveTo study the dynamic state changes of Hephaestin in cerebral cortex, corpus striatum and hippocampus after cerebral ischemia reperfusion injury and the relationship between it and reperfusion time. Preliminary discussion on the effectiveness in brain iron metabolism of Hephaestin.MethodHere to selective 40 clearing Wistar rat, and the embolism reperfusion model was divided randomly into 8 groups, control group(5), sham operation group(5), ischemic reperfusion group(30), and the last groups were divided randomly into 6 subsections,every subsections has 5 rats,that is 4h,8h,16h,32h, 64h,128h after ischemic reperfusion.The ischemic refusion model was established, immunohistochemistry staining, and the Leica Q550IW system was used to analyze the picture,the mean optical density values in cerebral cortex, corpus striatum was worked out in different reperfusion time.ResultHephaestin was expressed in cerebral cortex,corpus striatum in normal rat.After 8 hours of cerebral ischemia reperfusion,the expression of Hephaestin was increasing obviously,and it could last out 64 hours after reperfusion.At 16 to 32 hours after reperfusion it get to peak(P<0.05),and at 128 hours it was diminished greatly (P<0.05).ConclusionThe change of expression of hephaestin in the cortex and striatum during acute cerebral ischemia and reperfusion was greatly.From the study we can see the expression of Hephaestin was most obviously after reperfusion 16hour to 32hour,and after reperfusion 128hour the expression appeared the lowest leval. and the change may play an important role in the pathphysiology after acute cerebral ischemic.It is possible that the expression of Hephaestin is closely related to the degree of injury of brain tissue and the level of iron in the cerebral tissue after cerebral injury.