醒脑静注射液对缺氧缺血性脑损伤大鼠S100B蛋白及低氧诱导因子-1α和血管内皮生长因子表达的影响

Effect of Xingnaojing on the expressions of S100B,hypoxia inducible factor-1α and vascular endothelial growth factor in rats with hypoxic-ischemic brain damage

目的探讨缺氧缺血性脑损伤(hypoxic ischemic brain damage,HIBD)大鼠应用醒脑静注射液治疗后血清S100B蛋白、低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达水平变化及意义。方法 7日龄Wistar大鼠96只,随机分为假手术组、HIBD组、醒脑静预处理组和醒脑静治疗组各24只。假手术组仅分离左颈总动脉不结扎,不作低氧处理;余3组分离左侧颈总动脉并结扎,制作HIBD大鼠模型。醒脑静预处理组于制备模型前腹腔注射醒脑静注射液10mL/kg,连续3d;醒脑静治疗组制备模型即刻腹腔注射醒脑静注射液10mL/kg,1次/d,连续注射至处死日;HIBD对照组制备模型即刻腹腔注射等量生理盐水。各组分别于模型制备后1、3、5、7d断头处死大鼠各6只,取脑组织RNA,采用实时荧光定量PCR法检测各组脑组织S100B,HIF-1α和VEGF mRNA表达水平。结果模型制备后1、3、5、7d,醒脑静预处理组S100BmRNA(1.4±0.2、2.6±0.4、3.0±0.3、1.6±0.2)、醒脑静治疗组S100B(1.3±0.1、2.1±0.2、1.7±0.2、1.4±0.3)明显低于HIBD组(1.7±0.2、2.8±0.3、3.6±0.5、2.2±0.5)(P<0.05),醒脑静预处理组HIF-1αmRNA(1.6±0.1、2.2±0.2、3.3±0.2、4.5±0.1)和VEGF mRNA(1.3±0.1、2.4±0.2、3.3±0.2、4.6±0.1)与醒脑静治疗组HIF-1αmRNA(1.7±0.2、4.1±0.1、5.6±0.1、7.3±0.2)和VEGF mRNA(1.6±0.1、3.1±0.2、5.2±0.2、6.4±0.3)均高于HIBD组[HIF-1αmRNA(1.5±0.2、1.8±0.1、2.2±0.2、2.8±0.2),VEGF mRNA(1.2±0.1、1.7±0.1、2.5±0.2、3.1±0.2)](P<0.05);以上数据均高于假手术组(P<0.05)。结论醒脑静注射液可上调HIBD大鼠HIF-1αmRNA和VEGF的mRNA表达,下调S100BmRNA表达。

Objective To investigate the effect of Xingnaojing on the expressions of S100B, hypoxia inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF） in rats with hypoxic-ischemic brain damage （HIBD）. Methods Ninety-six 7-day-old rats were randomly divided into sham-operation group, HIBD group, Xingnaojing preconditioning group and Xingnaojing treatment group, with 24 rats in each group. Sham-operation group was treated with left-commoncarotid-artery separation, without ligation and hypoxia. The other three groups were treated with left-common-carotid- artery separation and ligation to establish HIBD rat models. Xingnaojing preconditioning group was injected 10 mL/kg Xingnaojing for 3 days before the establishment of HIBD models. Xingnaojing treatment group was injected 10 mL/kg Xingnaojing immediately after the establishment of HIBD models once a day till the rats were sacrificed. HIBD group was intraperitoneally injected the same volume of normal saline. Six rats from each group were sacrificed by day 1, 3, 5 and 7 after the establishment of HIBD models to obtain the RNA of brain tissue. The expressions of S100B, HIF-1v and VEGF mRNA were detected by using fluorescent quantitation PCR in all groups. Results The levels of S100B mRNA were significantly lower in Xingnaojing preconditioning group （1.4±0.2, 2.6±0.4, 3.0±0.3, 1.6±0.2） and Xingnaojing treatment group （1.3±0.1, 2.1±0.2, 1.7±0.2, 1.4±0.3） than those in HIBD group （1.7±0.2, 2.8±0.3, 3.6±0.5, 2.2±0.5） （P〈0.05）, and the levels of HIF-1α mRNA and VEGF mRNA were significantly higher in Xingnaojing preconditioning group （HIF-1α mRNA： 1.6±0.1, 2.2±0.2, 3.3±0.2, 4.5±0. 1; VEGFmRNA： 1.3±0.1, 2.4±0.2, 3.3±0.2, 4.6±0.1） and in Xingnaojing treatment group （HIF-1α mRNA： 1.7±0.2, 4.1±0.1, 5.6±0.1, 7.3±0.2; VEGFmRNA： 1.6±0.1, 3.1±0.2, 5.2±0.2, 6.4±0.3） than those in HIBDgroup （HIF-1α mRNA： 1.5±0.2, 1.8±0.1, 2.2±0.2, 2.8±0.2; VEGF mRNA： 1.2±0.1, 1.7±0.1, 2.5±0.2, 3.1±0.2） by day 1, 3, 5 and 7 after the establishment of HIBI） models （P%0.05）, which were all significantly higher than those in sham-operation group （P〈0.05）. Conclusion Xingnaojing injection can up-regulate HIF-1α and VEGF mRNA expression, and downregulate S100B mRNA expression in HIBD rats.