丁苯酞依赖Rho激酶/组织因子途径减弱内皮中性粒细胞的聚集来改善脑缺血/再灌注损伤  

Improvement of cerebral ischemia/reperfusion injury via butylphthalein dependent Rho kinase/tissue factor pathway attenuates neutrophil aggregation

作　　者：袁波[1] 谭莉[1] 李晓辉[1] 万芳超 李鑫[1] YUAN Bo;TAN Li;LI Xiao-hui;WAN Fang-chao;LI Xin(Department of Neurology,Hunan University of Chinese Medicine,Changsha 410208,Hunan,China)

机构地区：[1]湖南省脑科医院神经内科,湖南长沙410007

出　　处：《生物医学工程与临床》2021年第4期407-414,共8页Biomedical Engineering and Clinical Medicine

基　　金：湖南省教育厅科学研究项目(18C0406)。

摘　　要：目的明确丁苯酞是否依赖Rho激酶/组织因子(TF)途径减弱内皮中性粒细胞的聚集来改善脑缺血/再灌注损伤。方法选择SPF级健康雄性ICR小鼠40只,鼠龄11~12周龄,体质量20~30 g。随机(随机数字法)分为空白组、模型组、单纯3-正丁基苯酞(NBP)治疗组,NBP+TF抑制组。建立缺血/再灌注损伤模型,单纯NBP治疗组、NBP+TF抑制组造模后予以NBP注射液[10 mg/(kg·d)]连续腹腔注射7 d。空白组、模型组均予以等量0.9%氯化钠溶液(生理盐水)腹腔注射7 d。造模后评估各组给药前后神经功能缺损评分[神经严重缺损评分(NSS)]、血压变化;苏木精-伊红(HE)染色光学显微镜下观察脑组织结构损伤程度,用Western blot检查Rho激酶的相对活性,聚合酶链式反应(PCR)技术检测TF mRNA的水平,酶联免疫吸附分析(ELISA)法检测嗜中性粒细胞趋化因子(CINC)表达水平。结果与模型组比较,单纯NBP治疗组缺血再灌注小鼠脑组织的病理损伤明显改善,血压降低[复苏2 h:(127.18±3.24)mmHg vs(128.25±2.85)mmHg;给药7 d:(88.68±8.68)mmHg vs(113.68±3.44)mmHg],Rho激酶活性(0.34±0.10 vs 0.64±0.14)及TF mRNA表达降低(0.40±0.12 vs 0.96±0.18),内皮细胞损伤引起的炎症反应减弱[CINC含量:(14.84±2.39)pg/mL vs(43.68±3.25)pg/mL],显著改善神经功能缺损症状[NSS:造模苏醒时,(9.40±2.01)分vs(9.30±1.84)分;给药7 d,(6.00±2.08)分vs(8.00±1.82)分](P<0.05)。与模型组比较,NBP+TF抑制组大鼠中Rho激酶活性也能降低(0.57±0.15 vs 0.64±0.14),血压降低[给药7 d:(100.56±3.27)mmHg vs(88.68±8.68)mmHg],但对内皮-嗜中性粒细胞相互作用的抑制作用不明显[CINS含量:(35.25±2.51)pg/mL vs(43.68±3.25)pg/mL],脑组织结构不完全性损伤,神经功能缺损症状改善不明显[NSS:造模苏醒时,(9.50±1.96)分vs(9.30±1.84)分;给药7 d,(7.10±1.25)分vs(8.00±1.82)分](P>0.05)。结论NBP依赖Rho激酶/TF途径减弱内皮-嗜中性粒细胞相互作用,来改善脑缺血/再灌注损伤。Objective To investigate whether butylphthalide attenuates endothelial neutrophil aggregation via Rho kinase/tissue factor(TF)pathway to improve cerebral ischemia/reperfusion injury.Methods Forty SPF-grade healthy male ICR mice were selected,which aged 11-12 weeks old with body weight of 20-30 g.The mice were divided into blank group,model group,pure 3-n-butylphthalide(NBP)treatment group and NBP+TF inhibition group.The model of ischemia/reperfusion injury was established,and butylphthalide injection[10 mg/(kg·d)]was given intraperitoneally for 7 days after modeling in NBP treatment group and NBP+TF inhibition group.Blank group and model group were given same amount of 0.9%sodium chloride solution injection(normal saline)for 7 days.After modeling,neurological deficit scores[neurological severity scores(NSS)]and blood pressure changes of each group before and after administration were evaluated.The hematoxylin-eosin(HE)staining optical microscope was used to observe brain tissue structural damage,Western blot was used to test relative activity of Rho kinase,polymerase chain reaction(PCR)method was used to detect TF levels and enzyme-linked immunosorbent assay(ELISA)to detect expression level of neutrophil chemotactic factor(CINC).Results Compared with model group,the pathological damage of ischemia/reperfusion mice in NBP treatment group was significantly improved,and the blood pressure decreased[recovery 2 hours:(127.18±3.24)mmHg vs(128.25±2.85)mmHg;administration 7 days:(88.68±8.68)mmHg vs(113.68±3.44)mmHg],Rho kinase activity(0.34±0.10 vs 0.64±0.14)and TF mRNA expression(0.40±0.12 vs 0.96±0.18)were reduced,endothelial cell injury and inflammation were weakened[CINC content:(14.84±2.39)pg/mL vs(43.68±3.25)pg/mL],and symptoms of neurological deficit were significantly relieved[NSS:at the time of model recovery,(9.40±2.01)scores vs(9.30±1.84)scores;administration 7 days,(6.00±2.08)scores vs(8.00±1.82)scores](P<0.05).Compared with NBP treatment group,in NBP+TF inhibition group,the activity of Rho kin

关 键 词：丁苯酞 脑缺血/再灌注损伤 Rho激酶/TF途径通路 

分 类 号：R965[医药卫生—药理学] R743[医药卫生—药学]