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Moxa floss is the main material of moxibustion, which plays a therapeutic role through the thermal effect of combustion. In this paper, TG/DSC thermal analysis method was used to study the characteristic parameters of combustion heat of moxa floss produced in Qichun, and the thermal therapeutic effect and mechanism of moxibustion were studied through moxibustion OA animal model. The results show that the combustion process of moxa floss can be divided into three stages: drying, combustion oxidation and carbonization, and ash burnout. The combustion properties of moxa floss are between herbaceous and woody, and tend to be woody, with flammable, slow and lasting combustion characteristics. Moxibustion can relieve the pathological state of knee joint synovium to a certain extent, reduce knee joint swelling and blood stasis in OA rats, reduce interstitial edema, and improve local inflammation. The mechanism and target point of moxibustion treatment for OA may be up-regulating TRPM3 gene to activate ion channels, affecting calcium metabolism and reducing OA swelling degree; down-regulation of GAPDH affects glucose metabolism of knee synovial cells and mediates anti-inflammatory effect. Down-regulation of pain-related gene MMP24 is helpful to relieve OA pain. Up-regulation of CTNNB1 activates Wnt/β-catenin signaling pathway and affects differentiation and regeneration of OA chondrocytes. This study reveals the pyrolysis characteristics of moxa floss for the first time and discusses the biological effect and possible mechanism of moxibustion heat, providing new ideas for the quality evaluation of moxa floss and the mechanism of moxibustion therapy.