从外周TLR4/NF-κB信号通路至中枢正负调节介质探究退热六法对发热幼兔的退热效果及机制研究

前期研究主要结论如下：①小儿推拿退热安全、有效；②小儿推拿对两种类型（细菌和病毒）、三个年龄段（婴儿、幼儿、学龄前）、三种热度（低、中、高）的疗效亟待评价、机制亟须阐述。基于以上，实验分疗效评价和机制研究。在疗效评价中，拟采用退热六法对发热幼兔进行干预，通过血常规联合流式细胞术检测血细胞的形态学变化，行为学联合形态学作为疗效评价指标，评价退热六法的退热效果。在机制研究中，拟从外周至中枢的体温调节通路（血清→脑脊液→下丘脑），采用分子生物学技术检测外周TLR4/NF-κB信号通路关键指标（TLR4、NF-κB、IL-1β、IL-6、TNF-α、IFN等）、中枢POAH区正调节介质（PGE2、cAMP等）、腹中隔区负调节介质（AVP、α-MSH、β-EP等）的表达。对结果综合分析，得出退热六法对于两种致热类型、三个年龄段、三种热度的退热疗效及机制，为小儿推拿治疗发热提供科学依据。

The primary conclusions of preliminary research are as follows: ① infant tuina for the treatment of fever is safe and effective; ② the effectiveness and the mechanism of infant tuina for the use of two different types (bacterial and viral), three different age groups (Infants, toddlers, preschool age), and three different degrees of fever (low, medium, high) are urgently in need of elucidation. Based on the above, this experiment is divided into an evaluation of effectiveness and research of the mechanism. In the evaluation of the effectiveness, this research project will use ‘reducing fever six methods’ technique on New Zealand infant rabbits as the intervention. Through standard blood analysis combined with flow cytometry, the morphological changes of blood cells will be detected; ethological and morphological methods will be combined to evaluate the therapeutic effectiveness, and to evaluate the effectiveness of using infant tuina to treat fever in an experimental setting. To study the mechanism, the body temperature regulation pathway (serum→cerebrospinal fluid→hypothalamus) from the periphery to the central region will be studied to explore the mechanism of the therapeutic effectiveness. Integrated molecular biology will be used to detect the expression of peripheral TLR4/NF-κB signaling pathway key indicators (TLR4、NF-κB、IL-1β、IL-6、TNF-α、IFN, etc.), the role of central POAH area in adjusting the medium (PGE2, cAMP, etc.), and the negative regulation of the ventral septal area (AVP, alpha MSH, beta EP, etc.). A comprehensive analysis of the results will be carried out to clarify the antipyretic effect and mechanism of‘reducing fever six methods’for the use of two different types，three different age groups，and three different degrees of fever, as well as provide scientific evidence on the safety of using infant tuina to reduce fever.