NOTCH抑制剂DAPT对颅脑损伤大鼠神经保护作用及其机制  

Neuroprotective mechanism of notch inhibitor DAPT on brain injury in rats

作　　者：张秋峰[1] 卢广平[2] 赵春艳 Zhang Qiufeng;Lu Guangping;Zhao Chunyan(Shangqiu First People′s Hospital,Department of Emergency Intensive Care,Xuzhou Medical University Shangqiu Clinical College,Shangqiu 476000,China;Department of Emergency,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Emergency,Shangqiu First People′s Hospital,Shangqiu 476000,China)

机构地区：[1]商丘市第一人民医院,徐州医科大学商丘临床学院急诊重症科,476000 [2]郑州大学第一附属医院急诊科,450000 [3]商丘市第一人民医院急诊科,476000

出　　处：《中华实验外科杂志》2021年第6期1093-1095,共3页Chinese Journal of Experimental Surgery

摘　　要：目的:探讨NOTCH抑制剂DAPT对颅脑损伤大鼠神经保护作用及其机制。方法:60只SD大鼠随机分为假手术组、模型组和DAPT组。模型组和DAPT组大鼠采用自由落体法建立颅脑外伤模型,假手术组不做颅脑损伤。DAPT组大鼠经颅内注射DAPT 50μg/只,假手术组和模型组大鼠经颅内注射等体积生理盐水;治疗2周后,采用神经行为学评分评价3组大鼠神经功能状态;采用蛋白质免疫印迹(Western blot)分析3组大鼠脑组织Notch通路蛋白表达水平;采用酶联免疫吸附实验(ELISA)分析3组大鼠脑组织炎性因子水平;采用活性氧(ROS)试剂盒和JC-1试剂盒检测3组大鼠脑组织ROS水平和线粒体膜电位。组间比较采用单因素方差分析。结果:DAPT组大鼠神经功能评分[(4.81±0.61)分]明显低于模型组[(8.79±1.88)分],差异有统计学意义(t=3.110,P<0.05)。DAPT组大鼠脑组织Notch1、Delta 1和NICD表达水平(0.76±0.18、0.99±0.14、0.64±0.15)明显低于模型组(1.64±0.27、2.09±0.35、1.42±0.11),差异有统计学意义(t=2.528、2.107、2.081,P<0.05)。DAPT组大鼠脑组织炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6和IL-1β水平[(98.12±7.65)、(78.22±8.19)、(69.28±7.25)pg/ml]明显低于模型组[(198.43±12.65)、(128.44±10.38)、(98.55±5.90)pg/ml],差异有统计学意义(t=7.120、5.891、4.882,P<0.05)。DAPT组大鼠脑组织ROS水平(501.44±29.45)明显低于模型组(747.33±45.28),差异有统计学意义(t=6.912,P<0.05)。DAPT组大鼠脑组织线粒体膜电位(17.35±2.76)明显高于模型组(10.38±2.09),差异有统计学意义(t=4.817,P<0.05)。结论:DAPT可显著抑制Notch信号通路,降低颅脑损伤大鼠脑组织炎性因子、ROS和提高线粒体膜电位,进而保护颅脑损伤大鼠的神经功能。Objective To investigate the neuroprotective mechanism of notch inhibitor DAPT in rats with traumatic brain injury.Methods Totally,60 SD rats were randomly divided into sham operation group,model group and DAPT group.The rats in the model group and DAPT group were used to establish the model of craniocerebral injury by free fall,and the sham operation group was not used for craniocerebral injury.The rats in DAPT group was injected into the brain with 50μg DAPT.The rats in sham operation group and model group were injected with the same volume of normal saline.After 2 weeks of treatment,neurobehavioral score was used to evaluate the neurological function of the three groups.The expression level of Notch pathway protein in the three groups was analyzed by Western blotting.The levels of inflammatory factors in the three groups were analyzed by Enzyme linked immunosorbent assay(ELISA).The level of ROS and mitochondrial membrane potential were analyzed by reactive oxygen species(ROS)kit and JC-1 kit.The difference was statistically significant when P<0.05.Results The neurological function score in DAPT group(4.81±0.61)was significantly lower than that in the model group(8.79±1.88)(t=3.110,P<0.05).The expression levels of Notch1,Delta 1 and NICD in DAPT group(0.76±0.18,0.99±0.14 and 0.64±0.15 respectively)were significantly lower than those in the model group(1.64±0.27,2.09±0.35 and 1.42±0.11 respectively,t=2.528,2.107,2.081,P<0.05).Tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1 levels in DAPT group[(98.12±7.65),(78.22±8.19)and(69.28±7.25)pg/ml respectively]were significantly lower than those in the model group[(198.43±12.65),(128.44±10.38)and(98.55±5.90)pg/ml respectively,t=7.120,5.891,4.882,P<0.05].The level of ROS in DAPT group(501.44±29.45)was significantly lower than that in the model group(747.33±45.28,t=6.912,P<0.05).The mitochondrial membrane potential in DAPT group(17.35±2.76)was significantly higher than that in the model group(10.38±2.09,t=4.817,P<0.05).Conclusion DAPT can sig

关 键 词：颅脑损伤 神经功能 炎症 活性氧 

分 类 号：R651.15[医药卫生—外科学]