The Mechanism Study Of External Therapy For Knee Osteoarthritis Based On ‘Warming Channels And Activating Blood Circulation,Truncating And Reversing’ Through The ThemoTRPs-mediated Cold Hyperalgesia

Knee Osteoarthritis is the most common degenerative disease in orthopaedics,however there is no specific therapeutic schedule to treat KOA at present due to the unclear pathogenesis.Pain is the earlist symptom and has the most profound impact-patients who suffer from pain are unwilling to take joint functional training that can promote metabolism of peripheral tissues of the joint and aerobic exercise which can help control body weight,this has shaken patients’ confidence towards conservative treatment and has a negative impact on the self-assessment after surgery.Pain caused by KOA is sensitive to cold stimulus,recent researches revealed that this is a kind of cold stimulus hyperalgesia.TRPA1 and TRPM8 can mediate pain signal transduction when exposed to cold stimulus.In order to analyze KOA pain effectively and develop targeting drugs,it’s essential to find out whether TRPA1 and TRPM8 are involved in KOA,and whether they are related to the pharmaco-mechanism of Layers Adjusting External Application,a distinctive preparation in our hospital that has therapy effect on alleviating the pain caused by KOA,is also the concern of this subject.Objective:Explore the connection between TRPA1,TRPM8 and cold stimulus hyperalgesia of KOA,and intervention effect and pharmaco-mechanism that YiCeng emplastrum has on cold stimulus hyperalgesia of KOA.Methods:1.Compare protein and mRNA content of TRPA1 and TRPM8 in human synovial tissues of KOA(abandonded synovium from total knee anthroplasty)with non-KOA(synoxial debris from arthroscopic surgery after meniscus injury).2.Compare pain threshold of cold stimulus and protein and mRNA content of TRPA1 and TRPM8 of KOA rats model,impact that selective blockers of TRPA1,TRPM8 have on cold pain threshold and on electrophysiological properties of TRPA1 and TRPM8 in synoviocyte.3.Investigate effects of Layers Adjusting External Application on pain threshold of cold stimulus and protein and mRNA content of TRPA1 and TRPM8 of synovial tissues in KOA rats model.4.Investigate effects of Layers Adjusting External Application on the content of KOA inflammatory cytokines as inflammatory cytokines regulate the maintenance of activation of TRPA1 and TRPM8.Results:1.By comparison,protein and mRNA content of TRPA1 and TRPM8 in human synovial tissues of KOA was more than that in non-KOA with statistical significance(p<0.05).2.The pain Paw withdrawal latency by cold stimulus of KOA rats declines significantly compared to the healthy blank control group and the protein and mRNA content of TRPA1 and TRPM8 increases with statistical significance(p<0.05).Selective blockersof TRPA1,TRPM8 can up-regulate the Paw withdrawal latency in KOA rats by blocking calcium ion influx into current via TRPA1 and TRPM8 in synoviocyte with statistical significance(p<0.05).3.Layers Adjusting External Application can up-regulate the pain threshold of cold stimulus of KOA rats and down-regulate protein and mRNA content of TRPA1 and TRPM8 of synovial tissues in KOA rats with statistical significance(p<0.05).4.Layers Adjusting External Application can down-regulate content of inflammatory cytokines and up-regulate content of anti-inflammatory cytokines in KOA rats as it can inhibit the activation of TRPA1 and TRPM8 channels.Conclusions:TRPA1 and TRPM8 are important mediators of cold stimulus hyperalgesia of KOA,Layers Adjusting External Application may exert analgesic effect by down-regulating the expression of TRPA1 and TRPM8 and inhabit the activity of TRPA1 and TRPM8 channels.External therapy for knee osteoarthritis based on’warming channels and activating blood circulation,truncating and reversing’ is extremely valuable.