抑制可溶性环氧物酶对局灶性脑缺血再灌注大鼠的神经保护作用  

Neuroprotective effect of inhibition of soluble epoxide hydrolase on focal cerebral ischemia/repedusion in Rats

作　　者：原美艳[1] 朱雨岚[1] 

机构地区：[1]哈尔滨医科大学附属第二临床医学院神经内科,150086

出　　处：《国际脑血管病杂志》2013年第2期108-113,共6页International Journal of Cerebrovascular Diseases

摘　　要：目的探讨可溶性环氧物酶（soluble epoxide hydrolase，sEH）抑制剂12-（3-金刚烷-1-基脲基）-十二烷酸[12—（3-adamantan-1-yl—ureido）-dodec—anoicacid，AUDA]对局灶性脑缺血再灌注大鼠的神经保护作用和机制。方法60只雄性Sprague—Dawley大鼠随机分为假手术组、生理盐水对照组以及小剂量（0．157ml／kg）、中剂量（0．235ml／kg）和大剂量（0．314ml／kg）AUDA处理组（每组12只），各组随机取4只大鼠分别用于梗死体积、细胞凋亡和p-Akt免疫组织化学检测。线栓法建立大脑中动脉闭塞再灌注模型。各AUDA处理组和生理盐水对照组均在再灌注前分别经腹腔给予相应剂量的AUDA或等体积生理盐水。再灌注24h时进行神经功能缺损评分。2,3,5-氯化三苯基四氮唑染色法检测脑梗死体积。原位缺口末端标记法（TdT—mediated dUTP nick end labeling,TUNEL）检测梗死周围区脑组织细胞凋亡。免疫组织化学法检测梗死周围区脑组织p-Akt表达。结果TTC染色显示，假手术组未见梗死。生理盐水对照组以及小剂量、中剂量和大剂量AUDA组梗死体积分别为（254．146±25．481）、（212．679±7．514）、（150．188±33．997）和（99．563±3．415）mm3，存在显著性差异（F=39．637，P=0．000）。各剂量AUDA组均显著性小于对照组（P均=0．000）。中剂量AUDA组显著性小于小剂量AUDA组（P=0．002），而大剂量AUDA也显著性小于小剂量AUDA组（P=0．000）和中剂量AUDA组（P=0．006）。TUNEL染色法显示，假手术组仅可见少量凋亡细胞[（6．400±1．477）个／高倍视野]。生理盐水对照组以及小剂量、中剂量和大剂量AUDA组凋亡细胞数量分别为（57．550±13．067）、（47．030±8．423）、（34．530±4．393）和（26．400±2．683）个／N倍视野，各剂量AUDA组显著性少于生理盐水对照组（P均〈0．01），中剂量和大剂量AUDA组显著性少于小剂量AUDA组（P均�Object±ive To investigate the neuroprotective effect of soluble epoxide hydrolase （sEH） inhibitor 12-（3-adamantan-l-yl-ureido） dodecanoic acid （AUDA） on focal cerebral ischemiaJreperfusion in rats and its mechanisms. Methods Sixty male Sprague-Dawley rats were randomly divided into sham operation and saline control groups, as well as low-dose （0. 157 ml/kg）, medium-dose （0.235 ml/kg） and high-dose （0. 314 ml/kg） AUDA groups （n = 12 in each group）. Four rats in each group were selected for infarct volume, cell apoptosis and p-Akt immunohistochemistry detection. A model of middle cerebral artery isctaemia/ reperfusion was induced by the suture method. The corresponding dose AUDA or equal volume of saline was injected intraperitoneally before reperfusion in each AUDA group and the saline control group. Neurological deficit scores were performed at 24 h of reperfusion. 2,3,5 triphenyltetrazofium chloride （TTC） staining was used to detect infarct volume. TdT-medlated dUTP nick end labeling （TUNEL） was used to detect apoptotic cells of brain tissue in the pefiinfarction area. Immunohistochemical method was used to detect p-Akt expression of brain tissue in the pefi-infarction area. Results TTC staining showed no infarction was observed in the sham operation group. The infarction volumes in the saline control group as well as the low-dose, medium-dose and high-dose AUDA groups were 254. 146 ± 25. 481, 212. 679 ± 7. 514, 150. 188 ± 33. 997, and 99. 563 ± 3.415 mm3, respectively. There were significant differences （F = 39. 637, P = 0. 000）. The each dose AUDA group was significant less than the control group （all P = 0. 000）. The medium-dose AUDA group was sigfificantly less than the low-dose AUDA group （P = 0. 002）, and the high-dose AUDA group was also significantly less than the low-dose AUDA group （P = 0. 000） and medium-dose AUDA group （P = 0. 006）. TUNEL staining showed that a small number of apoptotic cells （6. 400 ± 1.477/high-power field） were obse

关 键 词：脑缺血 环氧水解酶类 酶抑制剂 细胞凋亡 原癌基因蛋白质c—akt 疾病模型 动物 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]