退热六法对脂多糖致热兔toll样受体4/核转录因子-κB及炎症因子的影响

Effects of“Six Methods of Antipyretic”on TLR4/NF-κB and inflammatory factors of LPS-induced fever rabbits

目的明确退热六法对脂多糖(lipopolysaccharide,LPS)致热幼兔的退热效果及抗炎机制。方法选取24只新西兰幼兔随机分为正常组、模型组、推拿组,每组8只,推拿组和模型组通过耳缘静脉注射0.5μg/kg LPS建立发热模型,1小时内肛温升高0.6℃为造模成功。推拿组于造模后1小时进行手法干预,干预方法为退热六法,即开天门、推坎宫、揉太阳、揉耳后高骨、清天河水、推脊,连续监测6小时肛温变化,比较造模前、造模后1小时、造模后3小时中性粒细胞比率变化,用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(Interleukin-1β,IL-1β)含量变化,用蛋白免疫印迹法(Western blot,WB)检测肝、肺组织toll样受体4(toll-like receptor 4,TLR4)、核转录因子-κB p65(nuclear factor-κB p65,NF-κB p65)表达含量。结果推拿后1小时、2小时、3小时,推拿组肛温显著低于模型组(P<0.05);造模后3小时,模型组中性粒细胞比率(neutrophil granulocyte,NEU%)高于造模前(P<0.05),推拿组NEU%显著低于模型组(P<0.05),推拿组血清中TNF-α、IL-1β含量低于模型组(P<0.05),推拿组肝、肺组织TLR4表达含量低于模型组(P<0.05),推拿组肝、肺组织NF-κB p65表达含量与模型组无明显差异(P>0.05)。结论退热六法降温作用明确,下调中性粒细胞比率、外周炎症因子TNF-α、IL-1β水平,从而影响TLR4信号通路实现调节体温作用,而这一作用不是通过NF-κB信号通路介导。

Objective To determine the antipyretic effect and anti-inflammatory mechanism of“Six Methods of Antipyretic”on LPS-induced fever in young rabbits.Methods 24 New Zealand rabbits were randomly divided into normal,model and Tuina groups(n=8 in each group).The model group and Tuina group were established by injecting 0.5μg/kg LPS into the auricular vein to establish fever model.A successful model was the anal temperature increased by 0.6℃within 1 hour.In the Tuina group,“Six Methods of Antipyretic”was performed 1h after modeling using,Kai tianmen,Tui kangong,Rou taiyang,Rou erhougaogu,Clear tianheshui and Tui Spinal.The change in anal temperature was monitored for six hours continuously.The ratios of neutrophils were compared before modeling,1h and 3h after modeling.The contents of serum TNF-αand IL-1βwere detected by ELISA,and the expression levels of TLR4 and NF-κB p65 in liver and lung tissues were detected by Western blot.Results The anal temperature in the Tuina group was significantly lower than that in the model group at 1h,2h and 3h after Tuina(P<0.05).At 3 h after modeling,the ratio of neutrophils(NEU%)in the model group was higher than that before modeling(P<0.05),and the NEU%in the Tuina group was significantly lower than that in the model group(P<0.05).The contents of TNF-αand IL-1βin the serum of Tuina group were lower than those in the model group(P<0.05).The expression levels of TLR4 in the liver and lung tissues of the Tuina group were lower than those in the model group(P<0.05),and the expression levels of NF-κB p65 in the liver and lung tissues of the Tuina group were not significantly different from those in the model group(P>0.05).Conclusion The antifebrile effect of“Six Methods of Antipyretic”is clear,and it can lower the neutrophil ratio,the levels of peripheral inflammatory factors TNF-αand IL-1β,thus affecting the TLR4 signaling pathway to achieve the effect of regulating body temperature,and this effect is not mediated by NF-κB signaling pathway.