AG490对大鼠脑损伤后血-脑屏障通透性及白细胞介素6和肿瘤坏死因子α表达的影响  

Impact of AG490 on blood-brain barrier permeability and expression of interleukin-6 and tumor necrosis factor-α after brain injury in rats

作　　者：张合鹏 杜爱玲[2] 李磊[1] 孟宪兵[3] 王振刚[2] 曹少鹏[1] 李广文[2] 季泰令[2] 

机构地区：[1]潍坊医学院外科学教研室,261000 [2]潍坊医学院附属医院神经科 [3]泰山医学院附属医院神经外科

出　　处：《中国脑血管病杂志》2015年第3期134-139,共6页Chinese Journal of Cerebrovascular Diseases

基　　金：山东省自然科学基金(ZR2011HM008)

摘　　要：目的探讨AG490对大鼠创伤性脑损伤(TBI)后血-脑屏障通透性以及炎性因子IL-6、TNF-α表达变化的影响。方法取健康成年雄性SD大鼠144只,按照随机数字表法分为对照组、创伤组、AG490干预组各48只,每组又分为4个亚组,分别对应伤后4 h、1 d、3 d、7 d时间点,每亚组各12只大鼠。应用液压冲击法制作脑创伤模型。应用伊文思蓝渗透法检测各组大鼠脑创伤后血-脑屏障通透性改变情况;采用实时荧光定量PCR方法检测大鼠脑组织中IL-6及TNF-αmRNA表达水平;免疫组化检测磷酸化酪氨酸激酶-2(P-JAK2)的表达。结果 (1)血-脑屏障通透性:脑损伤后4 h、1 d、3 d、7 d,创伤组血-脑屏障通透性增加[创伤脑组织伊文思蓝渗透量分别为(10.4±1.2)、(16.0±1.4)、(22.3±2.0)、(8.4±0.9)μg/g湿脑组织],与对照组相比,差异均有统计学意义(均P<0.01)。AG490干预组各时间段伊文思蓝渗透量分别为(9.1±1.0)、(12.8±1.1)、(17.5±1.4)、(7.1±0.8)μg/g湿脑组织,均明显低于创伤组(均P<0.01)。(2)IL-6 mRNA及TNF-αmRNA表达:脑创伤后4 h、1 d、3 d、7 d,创伤组IL-6 mRNA及TNF-αmRNA表达量分别为2.31±0.35、2.73±0.35、3.32±0.29、2.14±0.24和7.46±1.18、9.42±1.54、13.76±1.89、6.28±1.00,均明显高于对照组(均P<0.01);AG490干预组IL-6 mRNA及TNF-αmRNA表达量分别为1.14±0.22、1.54±0.23、1.94±0.32、1.26±0.21和5.57±0.88、7.78±1.02、11.51±1.29、5.05±0.97,均低于创伤组,但仍高于对照组,差异均有统计学意义(均P<0.01)。(3)P-JAK2的表达:脑创伤后各时间点,创伤组P-JAK2阳性细胞表达量均明显高于对照组(均P<0.01),分别为17.4±2.7、56.2±6.7、26.1±5.4、15.3±2.5;AG490干预组分别为12.2±1.4、41.5±4.6、19.4±4.1、9.6±2.0,均低于创伤组,但仍高于对照组,差异均有统计学意义(均P<0.01)。结论 TBI后急性期,AG490可通过抑制非受体型酪氨酸蛋白激酶/信号传导及转录激活因子信号通路,明显抑制脑组织炎性因Objective To investigate the impact of AG490 on the blood-brain barrier（ BBB）permeability and the expression of interleukin-6（ IL-6） and tumor necrosis factor-α（ TNF-α） after traumatic brain injury（ TBI） in rats. Methods A total of 144 healthy male SD rats were randomly divided into a control group,a trauma group,and an AG490 intervention group（ n = 48 in each group）. The rats in each group were redivided into four subgroups（ 4 h,1 d,3 d,and 7 d subgroups） according to the time points after cerebral injury（ n = 12 in each subgroup）. A brain trauma models were induced by hydraulic shock method. Evans blue was used to determine the changes of the BBB permeability after cerebral injury in each group. Real-time fluorescence quantitative PCR was to detect the expression levels of TNF-αand IL-6 mRNA in rat brain tissue. Immunohistochemistry was used to detect the expression of human phospho tyrosine kinase（ P-JAK2）. Results（ 1） The permeability of BBB： The permeability of BBB increased at 4 h,1 d,3 d and 7 d after brain injury in the trauma group（ Evans blue permeation： 10. 4 ±1. 2,16. 0 ± 1. 4,22. 3 ± 2. 0,and 8. 4 ± 0. 9 μg / g,respectively）. Compared with the control group,there were significant differences（ all P〈0. 01）. The Evans blue permeation of the AG490 intervention group were 9. 1 ± 1. 0,12. 8 ± 1. 1,17. 5 ± 1. 4 and 7. 1 ± 0. 8 μg / g,respectively at each time point,and they were all significantly lower than those of the trauma group（ all P〈0. 01）.（ 2） The expression of IL-6and TNF-α mRNA： The expression levels of IL-6 mRNA and TNF-α mRNA at 4 h,1 d,3 d and 7 d after traumatic brain injury in the trauma group were 2. 31 ± 0. 35,2. 73 ± 0. 35,3. 32 ± 0. 29,2. 14 ± 0. 24 and7. 46 ± 1. 18,9. 42 ± 1. 54,13. 76 ± 1. 89,and 6. 28 ± 1. 00,respectively,they were all significantly higher than those of the control group（ all P〈0. 01）. The expression levels of IL-6 mRNA and TNF-α mRNA of the AG490 intervention group were 1. 14

关 键 词：脑损伤 AG490 白细胞介素6 肿瘤坏死因子Α 血脑屏障 磷酸化酪氨酸激酶2 大鼠 

分 类 号：R651.15[医药卫生—外科学]