针刺负调控TLR4/TRIF信号通路关键因子对脑出血大鼠炎性反应表达的影响

目的探讨针刺“百会”透“曲鬓”穴对急性脑出血大鼠TLR4/TRIF信号通路关键因子Toll样受体4(TLR4)、TRIF(TIR-domain-containing adapter-inducing interferon-βTRIF)、TRAM(TRIF-related Adaptor Molecule,TRAM)、NF-κB(核因子κB)表达的影响。方法将72只Wistar雄性大鼠随机分为假手术组、模型组、针刺+模型组(简称针刺组)、TAK242组,每组按1 d、3 d、7 d时间节点分为3个亚组,每组6只。自体血注入法建立脑出血大鼠模型。采用针刺“百会”透“曲鬓”穴干预,Longa评分法对脑出血后大鼠神经功能进行评估;Western-blot法检测血肿组织TLR-4、TRIF、TRAM、NF-κB蛋白表达。结果神经行为学:与模型组比较,针刺组于3 d、7 d时间节点大鼠神经功能评分差异显著(P<0.05);TAK242组于3 d、7 d时间节点大鼠神经功能评分差异显著(P<0.01);与针刺组比较,TAK242组于3 d时间节点大鼠神经功能评分差异显著(P<0.05)。Western-blot结果:针刺组、TAK242组各时间节点TLR-4、TRIF、TRAM、NF-κB蛋白表达水平均显著低于模型组(P<0.01);TAK242组各时间节点TLR-4、TRIF、TRAM、NF-κB蛋白表达水平均显著低于针刺组(P<0.01)。结论针刺可能通过负调控TLR4/TRIF信号通路中TLR-4、TRIF、TRAM、NF-κB蛋白的表达,减轻脑出血后炎性损伤,改善大鼠神经功能缺损症状,发挥脑保护作用。

Effect of acupuncture negative regulation of key factors of TLR4/TRIF signaling pathway on expression of inflammatory response in rats with cerebral hemorrhage

Objective To investigate the effect of joined puncture of Baihui(GV 20)and Qubin(GB 7)on the expression of key factors TLR-4,TRIF,TRAM,NF-κB in TLR4/TRIF signaling pathway of rats with acute cerebral hemorrhage.Methods A total of 72 Wistar male rats were randomly divided into sham operation group,model group,acupuncture+model group(referred to as acupuncture group)and TAK242 group.Each group was divided into 3 subgroups of 1 d,3 d and 7 d,with 6 rats in each subgroup.The rat model of cerebral hemorrhage was established by autologous blood injection.Joined puncture of Baihui(GV 20)and Qubin(GB 7)was performed.Longa method was used to evaluate the neurological function of rats with cerebral hemorrhage and the protein expressions of TLR-4,TRIF,TRAM and NF-κB in the hematoma tissues were detected by Western-blot.Results Neuroethology:compared with the control group,the neurological function score in the acupuncture group was significantly different at 3d and 7d(P<0.05);while the neurological score of the TAK242 group at 3d and 7d was significantly different(P<0.01).Compared with the acupuncture group,the neurological function score of the TAK242 group was significantly different at 3d(P<0.05).Western-blot:TLR-4,TRIF,TRAM,NF-κB protein expression levels in the acupuncture group and the TAK242 group at each time point were significantly lower than those in the model group(P<0.01);and the protein expression levels of TLR-4,TRIF,TRAM,NF-κB in the TAK242 group at each time point were significantly lower than those in the acupuncture group(P<0.01).Conclusion Acupuncture can alleviate inflammatory injury after cerebral hemorrhage,and improve neurological deficit symptoms in rats and protect the brain,possibly by regulating the protein expression of TLR-4,TRIF,TRAM and NF-κB in the TLR4/TRIF signaling pathway.