脑缺血再灌注损伤中 FK506结合蛋白51对半胱氨酸蛋白酶－3及大鼠海马 CA1区神经元坏死的影响  

Effects of FKBP51 acting on Caspase-3 and rat hippocampal CA1 area neuronal necrosis in cerebral ischemia reperfusion injury

作　　者：王凯[1] 魏秀娥[1] 荣良群[1] 张清秀[1] 

机构地区：[1]徐州医学院第二附属医院神经内科,江苏省徐州221006

出　　处：《中国基层医药》2014年第24期3681-3685,共5页Chinese Journal of Primary Medicine and Pharmacy

基　　金：江苏省徐州市科技计划项目（XF10C030,2010）

摘　　要：目的：探讨脑缺血再灌注损伤中FK506结合蛋白51（FKBP51）对半胱氨酸蛋白酶－3（Caspase－3）及大鼠海马CA1区神经元坏死的影响。方法将清洁级SD大鼠随机分成假手术组（ Sham组）、缺血再灌注组（ I／R组）、溶剂对照组（ TE组）、FKBP51反义寡核苷酸组（ FKBP51 ASODN组）和FKBP51错义寡核苷酸组（ FKBP51 MSODN组），采用四血管阻塞模型大鼠全脑缺血模型造模。应用免疫印迹法测定FKBP51蛋白表达、FKBP51 ASODN对FKBP51蛋白水平的表达和Caspase－3活性的影响；采用苏木精－伊红溶液染色法测定FKBP51 ASODN对大鼠海马CA1区神经元坏死的影响。结果（1）在Sham组和I／R组（缺血再灌注0 min、15 min、30 min、1 h、3 h、6 h、1 d、3 d）中，FKBP51蛋白均有表达，并且表达量间的差异无统计学意义（F＝0．64，P＞0．05）。（2）FKBP51 ASODN组的FKBP51蛋白表达量明显减少，与Sham组差异有统计学意义（t＝8．21，P＜0．05）。（3）Sham组活化Caspase－3的表达量明显低于其他各组，差异有统计学意义（F＝12．31，P＜0．05）；与FKBP51 MSODN组比较，FKBP51 ASODN组的表达量相对较低，差异有统计学意义（ t＝9．71，P＜0．05）。（4）Sham组海马CA1区锥体细胞数量多[（186．3±2．5）个]并且排列密集，核大且圆，核仁明显；I／R 5 d组[（15．4±2．6）个]、TE组[（18．5±2．2）个]和FKBP51 MSODN组[（17．5±1．8）个]海马CA1区锥体细胞几乎完全消失，仅残留少数细胞，大量变性锥体细胞核固缩深染，胞浆嗜伊红，胞膜破裂，胞内容物释放， Sham组与其余各组之间的差异有统计学意义（χ^2＝81．91，P＜0．05）；FKBP51 ASODN组[（92．8±2．6）个]海马CA1区锥体细胞存活数增加，与其余各组的差异有统计学意义（χ^2＝52．36，P＜0．05）。结论在脑缺血再灌注损伤中，FKBP51可以增强活化的Caspase－3的表达并抑制神经元的存�Objective To explore the effect of FKBP51 acting on Caspase-3 and hippocampal CA1 area neu-ronal necrosis in cerebral ischemia reperfusion injury of rat.Methods SD rats were randomly divided into Sham group,ischemia reperfusion group （ I/R group ） , TE buffer group （ TE group ） , FKBP51 antisense oligonucleotide group （FKBP51 ASODN group） and FKBP51 missense oligonucleotide group （FKBP51 MSODN group）.Transient global cerebral ischemia rats models were made by four-vessel method.We used Western blot to detect the expression of FKBP51,the effect of FKBP51 ASODN to FKBP51 expression and Caspase-3 activity;while we used HE staining technique to detect FKBP51 ASODN effect to rat hippocampal CA1 area neuronal necrosis.Results （1） In Sham group and I/R group （0min,15min,30min,1h,3h,6h,1d,3d）,FKBP51 expressed,and the difference among the groups was no statistical significance （F=0.64,P〉0.05）.（2）The expression of FKBP51 in FKBP51 ASODN group was obviously reduced, and the difference was statistically significant compared with Sham group （ t =8.21, P 〈0.05）.（3）The expression of Cleaved-Caspase-3 in Sham group obviously declined than the other groups,the differ-ence between them was statistically significant （F=12.31,P〈0.05）;The expression of FKBP51 in FKBP51 ASODN＆amp;nbsp;group was decreasing compared with FKBP51 MSODN group,and the difference was statistical significance（t=9.71, P〈0.05）.（4）HE staining showed：the number of Sham group （186.3 ±2.5） hippocampal CA1 pyramidal cells was most.The cells arranged densely,and nucleoli were large and round,the difference was statistically significant com-pared with the other groups （χ^2 =81.91,P〈0.05）;The hippocampal CA1 pyramidal cells of I/R group （15.4 ± 2.6）,TE group （18.5 ±2.2） and FKBP51 MSODN group （17.5 ±1.8） were almost completely disappeared,only left a few residual cells,a great quantity of denaturated cells which presented karyopykosis,tinctorialed endochylema, ruptured of membran

关 键 词：全脑缺血 FK506结合蛋白51 反义寡核苷酸 半胱氨酸蛋白酶-3 ' 

分 类 号：R743.3[医药卫生—神经病学与精神病学]