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

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．...