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Through The Pretreatment To SD Rats By Atorvastatin, To Observe The Change Of Expression Of Apoptosis Of Neurons And TGF-β1after Cerebral Ischemia Reperfusion Injury

Posted on:2013-10-21Degree:MasterType:Thesis
Country:ChinaCandidate:R L GuanFull Text:PDF
GTID:2254330398984849Subject:Neurology
Abstract/Summary:PDF Full Text Request
Objective:To observe the expression of Fas,P53and Transforming growthfactorbeta-1(TGF-β1)on focal cerebral ischemia-reperfusion injury in SD rats; toobserve the influence of the expression of Fas、P53and TGF-β1on prophylactic useatorvastatin on focal cerebral ischemia-reperfusion injury in SD rats;to disscuss whetherwith atorvastatin can relieve the injury of endothelial function on focal cerebralischemia-reperfusion and possible mechanism.Methods:90healthy male SD rats,according to the principle of randomly dividedinto three groups(n=30):sham-operated group、control group and atorvastatin calciumpretreatment group (atorvastatin group to abbreviate), thirty rats in each group. Eachgroup was divided into5subgroups according to reperfusion2h(n=6)、6h(n=6)、24h(n=6)、48h(n=6)、72h(n=6) after ischemia for2hours.Atorvastatin regimencontinued rats,preoperative14days to atorvastatin regimen continued (10mg/kg/d) lineirrigation treatment stomach,once a day;sham-operated group and control group are inpreoperative14days in the stomach with volume and0.9%sodium chloride solution,once a day.Using the improved Longa-Zea method to establish ischemia Reperfusion modelin right cerebral middle artery of SD rat (each group was made ischemia-reperfusionmodele xcept the sham-operated group). Evaluate the neurological impairment score byreferring to Zea-Longa’s standards of five grades; all removed brains was stainedrespectively with2%Tripheny1Tetrazalium Chloride(2,3,5-Triphenyl tetrazaliumchloride, TTC); Judging the ischemic location, measuring the infarctionpathomorphology through Hematoxylin-eosin(HE);Observing he expression of Fas、P53and TGF-β1with immunohistochemical techniqu. Compare respectively:(1) Compare the difference beyween sham-operated group and control group inneurological impairment score,infarction volume appear,morphology of cerebral tissueand the expression of Fas、P53and TGF-β1.(2) Compare the difference beyween control group and atorvastatin group inneurological impairment score,infarction volume appear,morphology of cerebral tissueand the expression of Fas、P53and TGF-β1.(3) Observation the difference between the close two subgroup of control group inneurological impairment score,infarction volume appear,morphology of cerebral tissueand the expression of Fas、P53and TGF-β1.(4) Observation the difference between the close two subgroup of atorvastatingroup in neurological impairment score,infarction volume appear,morphology ofcerebral tissue and the expression of Fas、P53and TGF-β1.Results:1.Changes in neurologic impairment score:The neurological impairment score in control group was higher than that in thesham-operated group at every subgroup and the result has remarkable difference(P<0.01);Zea-Longa evaluation have no difference between the atorvastatin group andthe control group except in the group of24h、48h and72h reperfusion following2hcerebral ischemia(P<0.05).2.Changes in cerebral infarction volume:Sham-operated group have no infarction area;the infarction volume appear at2hcerebral ischemia and increased gradually after reperfusion in the control group and theatorvastatin group;compare with the difference between control group group andSham-operated group,the difference was remarkable(P<0.01);the infarction volumeshowed no difference between the atorvastatin group and the control group except in thegroup of24h、48h and72h reperfusion following2h cerebral ischemia(P<0.05).3.HE dyed observation:Sham-operated group have no infarction areas and evident pathological change;the number of neuron at ischemic area decreased, nervecells shrank anddegenerated,apparent edema between tissues and vacuolization in control group;compared with control group,the number of neuron which appared degeneration andnecrosis decreased,vacuolization and inter-tissue edema became relieved in atorvastatingroup. 4.Immunohistology result:(1)The changes of Fas protein expression:we can’t see any Fas cell expression in the sham-operated group.At each time ofreperfusion,the number of Fas positive cells in control group were higher than that insham-operated group,there has Significant differences (P<0.01);In control group andatorvastatin group,Fas cells increased since6h after reperfusion and reached the peak at48h after reperfusion,then decreased gradually,compare the time of the same group,24h and48h are higher than the others(P<0.05),compare with control group,thenumber of Fas positive cells in atorvastatin group reduced,the difference at every ofreperfusion (P<0.05),significant differences at24h and48h(P<0.01).(2) The changes of P53protein expression:A few of P53positive cells can be seen in sham-operated group.At each time ofreperfusion,the number of P53positive cells in control group were higher than that insham-operated group(P<0.05),In control group and atorvastatin group,P53cellsincreased since2h after reperfusion and reached the peak at48h after reperfusion,thendecreased gradually,compare the time of the same group,each time are higher than theothers(P<0.05),Compare with control group,the number of P53positive cells inatorvastatin group reduced,the difference at every of reperfusion (P<0.05),significantdifferences at24h (P<0.01).(3) The changes of TGF-β1protein expression:There has none TGF-β1cell expression in the normal cerebral tissue.At eachtime of reperfusion, the number of TGF-β1positive cells in control group weresignificant difference than that in sham-operated group(P<0.01).In control group andatorvastatin group, few TGF-β1cells expression at the2h and6h after reperfusion,increased since6h and reached the peak at24h,then decreased gradually at72h,compare the time of6h,24h are significant difference (P<0.01),there are little differentat the time of24h、48h and72h.Compare with control group,the number of TGF-β1positive cells in atorvastatin group increased,the difference at every of reperfusion(P<0.05),significant differences at24h、48h and72h(P<0.01).Conclusions:1.The expression of Fas、P53and TGF-β1can be increased in the process ofcerebral ischemia-reperfusion injury in SD rats.2.Prophylactic use atorvastatin could increase the expression of TGF-β1,reducedthe number of Fas and P53in the ischmia tissue in SD rats. 3.Prophylactic use atorvastatin could decrease focal cerebral ischemia-reperfusioninjury,relive the neurological functional defect,reduce infracted volume,relieve theneuronal degeneration,necrosis and tissue edema,the neuroprotective mechanism ofatorvastatin may be increase the expression of TGF-β1,reduced the content of Fas andP53on focal cerebral ischemia-reperfusion injury....
Keywords/Search Tags:cerebral, ischemia-reperfusion, atorvastatin Fas, P53, TGF-β1
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