补阳还五汤对小窝蛋白-1基因敲除小鼠脑缺血后炎性因子表达的影响  

Effects of Buyang Huanwu decoction on expressions of inflammatory cytokines after cerebral ischemia in Caveolin-1 knockout mice

作　　者：黄素芬[1] 易健[1] 周胜强[1] 罗东[1] 刘柏炎[1] 

机构地区：[1]湖南中医药大学,湖南长沙410007

出　　处：《中国中西医结合急救杂志》2016年第6期636-639,共4页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：国家自然科学基金（81273989）;湖南省自然科学基金（2015JJ2105）

摘　　要：目的 探讨小窝蛋白-1 （Cavl）在脑缺血后炎症反应中的作用及补阳还五汤对Cav1基因敲除小鼠脑缺血后脑组织肿瘤坏死因子-α（TNF-α）、细胞间黏附分子^-1（ICAM-1）表达水平的影响.方法 将Cav1基因敲除小鼠（KO）60只及同源野生小鼠（WT） 60只分别按随机数字表法分为假手术组、模型组、补阳还五汤组.采用大脑中动脉闭塞（MCAO）法复制小鼠局灶性脑缺血模型;制模后2h各组分别给予灌胃预处理,补阳还五汤组给予补阳还五汤煎液（由黄芪120 g,当归尾6 g,川芎3 g,红花3 g,赤芍4.5 g,桃仁3 g,地龙3 g组成）5 g/kg（临床等效剂量）,每日2mL;其他各组动物均给予等体积无菌生理盐水.于制模后3、7、10、14 d各组处死5只小鼠取脑组织,采用免疫组化法观察脑组织TNF-α、ICAM-1蛋白表达水平.结果 TNF-α、ICAM-1蛋白阳性细胞在整个大脑组织中均有表达,但主要表达在大脑皮质和海马区,TNF-α表达在细胞核、细胞质、细胞膜等整个细胞结构中,ICAM-1表达在细胞膜上.KO和WT两个假手术组小鼠脑组织均有TNF-α和ICAM-1蛋白的低水平表达;各时间点KO和WT两个模型组TNF-α、ICAM-1阳性细胞数均较同鼠种假手术组明显增多,制模后3d均明显增加[KO组：TNF-α（个/mm^2）为102.67±22.84比38.67±8.80,ICAM-1（个/mm^2）为47.33±3.89比19.33±3.14;WT组：TNF-α（个/mm^2）为70.00±17.72比45.83±10.53,ICAM-1（个/mm^2）为46.67±5.43比20.50±3.21],7d达高峰[KO组：TNF-α（个/mm^2）为127.50±18.06比34.83±8.77,ICAM-1（个/mm^2）为76.67±14.79比20.17±3.92;WT组：TNF-α（个/mm^2）为89.67±2.58比29.67±4.80,ICAM-1（个/mm^2）为48.33±8.09比15.50±1.52],10 d逐渐下降,14d达最低[KO组：TNF-α（个/mm^2）为99.67±18.33比34.00±8.10,ICAM-1（个/mm^2）为47.50±8.20比12.83±2.71;WT组：TNF-α（个/mm^2）为59.67±17.07比31.17±9.70,ICAM-1（个/mm^2）为44.33±7.15比19.67±3.56].KO和WT两个补阳还五汤组脑组�Objective To investigate the role of Caveolinl （Cavl） gene in the inflammatory response after cerebral ischemia and effects of Buyang Huanwu decoction on expressions of tumor necrosis factor-α （TNF-α ） and intercellular cell adhesion molecule^-1 （ICAM-1） after cerebral ischemia in Cavl gene knockout mice. Methods Carl knockout （KO） 60 mice and its homologous wild （WT） 60 mice were respectively divided into control group, model group and Buyang Huanwu decoction group according to random number table method. Middle cerebral artery embolism （MCAO） method was used to copy the model of focal cerebral ischemia in mice. Each group was given by gastric gavage pretreatment 2 hours after the modeling, Buyang Huanwu decoction group was given Buyang Huanwu decoction （astragalus membranaceus 120 g, angelica sinensis 6 g, rhizoma chuanxiong 3 g, safflower 3 g, radix 4.5 g, peach kernel 3 g and lumbricus 3 g） 5 g/kg （the clinical equivalent dose）, 2 mL per day, and all other animals were given equal volume of sterile saline. Five rats in each group were sacrificed on each of the following time points, the 3 rd, 7th, 10th, 14th day, and then the expressions of TNF-α and ICAM-1 protein in brain tissue were detected by immunohistochemistry. Results TNF-α and ICAM-1 protein positive cells expressed in the whole brain tissue, but mainly in the cerebral cortex and hippocampus region. TNF-α expression in the nucleus, cytoplasm, cell membrane and other cell structure, yet ICAM-1 in the cell membrane. The expression levels of TNF-α and ICAM-1 protein in the brain tissues of KO and WT sham-operated mice were all low. The numbers of TNF-α and ICAM-1 positive cells in KO group and WT model groups were significantly higher at each time point than those in sham-operated group （P 〈 0.01）, which were obviously increased on the 3rd day after modeling [KO group： TNF-α （pcs/mm^2） was 102.67±22.84 vs. 38.67±8.80, ICAM-1 （pcs/mm^2） was 47.33±3.89 vs. 19.33±3.14; WT group： TNF-α （pcs/m

关 键 词：小窝蛋白-1 补阳还五汤 脑缺血 肿瘤坏死因子-α 细胞间黏附分子-1 

分 类 号：R743[医药卫生—神经病学与精神病学]