活血化瘀方剂对重型颅脑损伤大鼠Wnt/β-连环蛋白信号通路表达的影响  

Effect of Huoxue Huayu decoction on Wnt/β-catenin signal pathway expression in rats with severe traumatic brain injury

作　　者：王景博 韦春珠 潘茂华 刘鑫杰 潘宇政[1] Wang Jingbo;Wei Chunzhu;Pan Maohua;Liu Xinjie;Pan Yuzheng(Department of Traditional Chinese Medicine,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Department of General Surgery,Shanglin People's Hospital,Nanning 530500,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院中医科,南宁530021 [2]上林县人民医院普通外科,南宁530500

出　　处：《中华危重病急救医学》2020年第9期1101-1106,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81460687);广西壮族自治区卫生厅中医药科技专项(GZLC14-29);国家中医药管理局"十一五"重点专科建设项目(20J1X1L402K214)。

摘　　要：目的动态观察重型颅脑损伤(sTBI)急性期大鼠神经功能缺损评分(NSS)、Wnt/β-连环蛋白(β-catenin)信号通路、脑源性神经营养因子(BDNF)与神经生长因子(NGF)的表达规律,探讨活血化瘀方剂的干预作用.方法将126只雄性SD大鼠按随机数字表法分为对照组(生理盐水2 kg/L)、模型组(生理盐水2 kg/L)、脑蛋白水解物组(BP组,5.6 g/kg)、桃红四物汤组(TH组,10.2 g/kg)、血府逐瘀汤组(XF组,15.6 g/kg)、通窍活血汤组(TQ组,9.6 g/kg)、补阳还五汤组(BY组,28.7 g/kg)7组,每组18只.采用改良Feeney自由落体法建立sTBI大鼠模型;对照组不予创伤处理.各组分别于伤后6 h灌胃相应药物,伤后1、3、7 d进行NSS评分;取大鼠海马组织,采用反转录-聚合酶链反应(RT-PCR)检测Wnt3a和β-catenin的mRNA表达;采用免疫组化法检测BDNF和NGF的阳性表达.结果与对照组比较,模型组大鼠伤后1 d即出现明显的颅脑损伤症状,表现为NSS评分明显升高,Wnt3a和β-catenin的mRNA表达明显上调,BDNF和NGF阳性表达明显增加,说明sTBI大鼠模型制备成功,并随时间延长呈现一定自我修复能力.与模型组相比,XF组、TQ组和BY组NSS评分于伤后1 d即明显下降(分:6.6±1.5、6.1±2.0、5.7±2.4比9.4±1.5,均P<0.05),而BP组及TH组NSS评分于伤后7 d才明显下降,且TQ组和BY组NSS评分下降幅度较其他药物组更加显著.与模型组比较,BP组海马组织Wnt3a和β-catenin的mRNA表达分别于伤后1 d、3 d明显升高,并随时间延长持续升高;4个活血化瘀方剂组Wnt3a和β-catenin的mRNA表达于伤后1~3 d均有不同程度的波动,但均于伤后7 d明显高于模型组,以BY组升高更为显著〔Wnt3a mRNA(2-ΔΔCt):154.7±4.1比17.4±1.0,β-catenin mRNA(2-ΔΔCt):17.05±0.45比2.74±0.13,均P<0.05〕,其次为TQ组〔Wnt3a mRNA(2-ΔΔCt):126.6±2.8比17.4±1.0,β-catenin mRNA(2-ΔΔCt):8.70±1.19比2.74±0.13,均P<0.05〕.与模型组比较,BP组BDNF和NGF的阳性表达在伤后1 d升高,但3 d达峰值后有Objective To observe the dynamic of neurological severity scores(NSS)and the expressions of Wnt/β-catenin signaling pathway,brain-derived neurotrophic factor(BDNF)and nerve growth factor(NGF)in rats with severe traumatic brain injury(sTBI),and to explore the effect of Huoxue Huayu decoction.Methods A total of 126 Sprague-Dawley(SD)rats were randomly divided into seven groups by random number table with 18 rats in each group,namely control group(normal saline 2 kg/L),model group(normal saline 2 kg/L),brain protolysate group(BP group,5.6 g/kg),Taohong Siwu decoction group(TH group,10.2 g/kg),Xuefu Zhuyu decoction group(XF group,15.6 g/kg),Tongqiao Huoxue decoction group(TQ group,9.6 g/kg)and Buyang Huanwu decoction group(BY group,28.7 g/kg).The sTBI rat model was reproduced by modified Feeney free fall method,and the rats in the control group were not treated with trauma.The rats in each group were intragastrical administered with corresponding drugs at 6 hours after injury,and the NSS scores were evaluated on the 1st,3rd and 7th days after injury.After the hippocampus was harvested,the mRNA expressions of Wnt3a andβ-catenin were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the positive expressions of BDNF and NGF were detected by immunohistochemistry.Results Compared with the control group,the rats in the model group showed obvious symptoms of craniocerebral injury at 1 day after injury,which was manifested as significantly increased NSS score,up-regulated mRNA expressions of Wnt3a andβ-catenin,and increased positive expressions of BDNF and NGF,which indicated that the sTBI rat model was successfully prepared and presented a certain self-repair ability with the extension of time.Compared with the model group,NSS scores in the XF group,TQ group and BY group significantly decreased at 1 day after injury(6.6±1.5,6.1±2.0,5.7±2.4 vs.9.4±1.5,all P<0.05);however,the NSS scores in the BP group and TH group decreased significantly at 7 days after injury,and the NSS scores in the TQ group a

关 键 词：桃红四物汤 血府逐瘀汤 通窍活血汤 补阳还五汤 重型颅脑损伤 Wnt/β-连环蛋白信号通路 神经营养因子 

分 类 号：R285.5[医药卫生—中药学]