Study on the antipyretic effect of pushing Tianheshui in young rabbits: focus on the α-MSH-mediated cAMP/PKA/NF-κB signaling pathway

基于α-MSH介导的cAMP/PKA/NF-κB信号通路探讨清天河水对幼兔的退热机制

Objective:To explore the antipyretic effect and partial mechanism of the pushing Tianheshui manipulation on lipopolysaccharide(LPS)-induced fever in young New Zealand rabbits.Methods:Thirty 50-day-old New Zealand rabbits were randomly assigned to five groups,including a normal group,a model group,a Tuina(Chinese therapeutic massage)group,a Tuina control group,and a drug group,with 6 rabbits in each group.All groups except for the normal group received LPS injections through the marginal ear vein to induce fever.One hour post-modeling,the Tuina and Tuina control groups received pushing Tianheshui manipulation and pushing manipulation on the medial middle of the hind limbs,respectively,administered every hour for a total of 3 interventions.The drug group was given acetaminophen oral liquid via gavage.Anal temperature was recorded every 30 min for 4.0 h to monitor temperature changes among groups.At 4.0 h post-modeling,hypothalamus samples from each group were analyzed using Western blotting(WB)and real-time quantitative polymerase chain reaction(RT-qPCR)to measure the relative expression levels ofα-melanocyte-stimulating hormone(α-MSH),melanocortin 4 receptor(MC4R),cyclic adenosine monophosphate(cAMP),protein kinase A(PKA),nuclear factor-κB p65(NF-κB p65),and interleukin(IL)-1βproteins and their mRNAs.Results:Compared to the model group,the Tuina group showed a significant reduction in the anal temperature from 3.5 h to 4.0 h post-modeling(P<0.05).The Tuina control group did not show a significant temperature reduction from 0.5 h to 4.0 h post-modeling(P>0.05).The drug group exhibited a significant temperature reduction from 1.5 h to 4.0 h post-modeling(P<0.05).At 4.0 h post-modeling,compared to the model group,the Tuina group showed significantly increased relative expression ofα-MSH and MC4R proteins and mRNAs(P<0.05)and significantly decreased relative expression of cAMP,PKA,NF-κB p65,and IL-1βproteins and mRNAs in the hypothalamus tissue(P<0.05).No significant differences were observed in these parameters in the Tuina control group compared to the model group(P>0.05).Conclusion:Pushing Tianheshui manipulation demonstrated a significant antipyretic effect,potentially linked to point specificity.Its mechanism may involve theα-MSH-mediated cAMP/PKA/NF-κB pathway.

目的:探讨清天河水推拿对脂多糖(LPS)致热新西兰幼兔的退热效果及部分作用机制。方法:将30只50日龄新西兰幼兔采用随机数字表法分为正常组、模型组、推拿组、对照推拿组和药物组,每组6只。除正常组外,其余组经耳缘静脉注射LPS溶液建立发热模型。造模1.0 h后,推拿组和对照推拿组分别行清天河水的推拿操作和后肢内侧正中推法操作,每1.0 h干预1次,共干预3次;药物组予以对乙酰氨基酚口服液灌胃。每30 min测量一次幼兔肛温,连续测量4.0 h以比较各组肛温变化。造模4.0 h后取各组幼兔下丘脑样本进行免疫印迹法(WB)和实时荧光定量聚合酶链反应(RT-qPCR)检测,比较各组α-黑素细胞刺激素(α-MSH)、黑皮质激素4型受体(MC4R)、环磷酸腺苷(cAMP)、蛋白激酶A(PKA)、核转录因子-κB p65(NF-κB p65)、白介素(IL)-1β蛋白及其mRNA的相对表达量。结果:与模型组比较,造模后3.5~4.0 h,推拿组肛温显著下降(P<0.05);造模后0.5~4.0 h,对照推拿组肛温无明显下降(P>0.05);造模后1.5~4.0 h,药物组肛温显著下降(P<0.05)。造模后4.0 h,与模型组比较,推拿组α-MSH、MC4R蛋白及mRNA相对表达量显著升高(P<0.05),cAMP、PKA、NF-κB p65、IL-1β蛋白及mRNA相对表达量显著降低(P<0.05)。与模型组比较,对照推拿组下丘脑组织α-MSH、MC4R、cAMP、PKA、NF-κB p65、IL-1β蛋白及mRNA相对表达量无显著变化(P>0.05)。结论:清天河水具有显著退热效果,且存在一定穴位特异性;其退热机制可能与α-MSH介导的cAMP/PKA/NF-κB通路有关。