通窍活血汤灌胃对小鼠颅脑损伤的改善作用及其机制  

Effect and mechanism of Tongqiao Huoxue decoction on neurological function recovery for mice with traumatic brain injury

作　　者：杜勇[1] 王革生[1] 张淑敏 马涛[1] 张少辉[1] 宋光荣 刘晓汉[3] 周玉嘉[1] DU Yong;WANG Gesheng;ZHANG Shumin;MA Tao;ZHANG Shaohui;SONG Guangrong;LIU Xiaohan;ZHOU Yujia(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;不详)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]民航总医院 [3]北京中医药大学

出　　处：《山东医药》2020年第9期48-52,共5页Shandong Medical Journal

基　　金：北京中医药大学青年教师项目(2018-JYBZZ-JS145);国家中医药管理局国家中医临床研究基地业务建设科研专项课题(JDZX2015043)。

摘　　要：目的观察通窍活血汤灌胃对小鼠颅脑损伤的改善作用,并探讨其机制。方法60只小鼠随机分为假手术组、模型组和治疗组各20只,模型组和治疗组小鼠采用改良自由落体法建立颅脑损伤模型。建模后6 h时,三组均给予凝血酶、甘露醇尾静脉注射,治疗组同时给予通窍活血汤灌胃。建模后24 h时,使用改良的神经功能缺损评分(mNSS评分)评估各组小鼠神经功能损伤程度。评估后每组取3只小鼠处死,取损伤侧脑组织,分别称取湿重和干重后,计算小鼠脑组织水含量。评估后每组取3只小鼠处死,取损伤侧脑组织,TUNEL法检测小鼠脑组织细胞凋亡率,PCR法检测小鼠脑组织线粒体DNA(mtDNA),Western Blotting法检测小鼠脑组织中自噬相关蛋白LC3-Ⅱ、LC3-Ⅰ、p62及锰超氧化物歧化酶(MnSOD)、细胞色素C氧化酶Ⅳ亚型(COXⅣ)、切割活化的半胱天冬酶-3(cleaved Caspase-3)、凋亡抑制基因Bcl-2蛋白。结果假手术组、模型组、治疗组mNSS评分分别为(2.00±0.47)、(15.00±0.47)、(6.00±0.94)分,组间相比,P均<0.05。假手术组、模型组、治疗组脑组织水含量分别为74.4%±0.36%、84.2%±0.64%和79.5%±0.70%,组间相比,P均<0.05。假手术组、模型组、治疗组细胞凋亡率分别为9.60%±1.87%、35.70%±6.61%和21.50%±5.19%,组间相比,P均<0.05。假手术组、模型组、治疗组脑组织mtDNA相对表达量分别为1.028±0.233、0.040±0.003和0.510±0.201,组间相比,P均<0.05。与假手术组相比,模型组和治疗组LC3Ⅱ/Ⅰ、MnSOD、COXⅣ、Bcl-2蛋白相对表达量均显著降低,p62、cleaved Caspase-3蛋白相对表达量均显著升高;与模型组相比,治疗组LC3Ⅱ/Ⅰ、MnSOD、COXⅣ、Bcl-2蛋白相对表达量均显著升高,p62、cleaved Caspase-3蛋白相对表达量均显著降低。结论通窍活血汤灌胃可改善小鼠颅脑损伤情况,其作用机制可能与激活线粒体自噬、抑制细胞凋亡有关。Objective To observe the effect of Tongqiao Huoxue decoction on traumatic brain injury mice and to explore its mechanism.Methods Sixty mice were randomly divided into the sham operation group,model group,and treatment group.The mice in the model group and the treatment group were modified with the free-fall method to establish the brain injury models.At 6 h after modeling,the mice in the three groups were given thrombin and mannitol via tail vein injection,while the mice in the treatment group were given Tongqiao Huoxue decoction at the same time.At 24 h after modeling,the neurological impairment of mice in each group was assessed by using the modified neurological deficit score(mNSS score).After the evaluation,3 mice in each group were sacrificed,and the injured lateral brain tissues were taken.The wet and dry weights of the mice were weighed and the water content in the brain tissues of the mice was calculated.After evaluation,3 mice in each group were sacrificed and the injured lateral brain tissue was taken.The TUNEL method was used to detect the apoptosis rate of mouse brain tissue cells.PCR was used to detect the mitochondrial DNA(mtDNA)in the mouse brain tissues.Western blotting was used to detect autophagy-related proteins LC3-Ⅱ,LC3-Ⅰ,p62 and manganese superoxide dismutase(MnSOD),cytochrome C oxidase subtype IV(COXⅣ),cleaved activated Caspase-3(cleaved Caspase-3),and Bcl-2 protein.Results The mNSS scores of the sham operation group,the model group,and the treatment group were(2.00±0.47),(15.00±0.47),and(6.00±0.94)points,respectively,with statistically significant differences between groups(all P<0.05).The water content in the brain tissues of the sham operation group,model group,and treatment group were 74.4%±0.36%,84.2%±0.64%and 79.5%±0.70%,respectively,with statistically significant differences between groups(all P<0.05).The apoptotic rates of the sham operation group,model group,and treatment group were 9.60%±1.87%,35.70%±6.61%and 21.50%±5.19%,respectively,with statistically significant

关 键 词：通窍活血汤 颅脑损伤 线粒体自噬 细胞凋亡 动物实验 

分 类 号：R285.5[医药卫生—中药学]