基于themoTRPs介导冷痛敏对“温经活血、截断扭转”外治膝骨关节炎的机理研究
发布时间：2019-05-09 18:06
【摘要】：膝骨关节炎(Knee Osteoarthritis,KOA)是最常见的骨与关节退行性疾病,因其发病机制一直未能明确,至今没有特效的治疗药物。KOA疼痛症状的出现很早,影响也最深远。KOA疼痛对冷温刺激非常敏感,近年来研究逐步揭示这是一类冷刺激痛敏。TRPA1和TRPM8是人体对低温刺激所致疼痛信号转递的介导者,膝骨关节炎中是否存在TRPA1、TRPM8的参与,对于认识KOA疼痛和开发靶向药物意义重大。"易层"贴敷膏剂是我院专利成方,以"温经活血"立法,通过"截断" KOA疼痛,"扭转" KOA病情进展,具有良好的效果,其药效机制是否与此有关,也是我们本课题研究的关注点。目的:探索TRPA1和TRPM8与KOA冷刺激痛敏之间的联系,以及温经活血外治("易层"贴敷膏剂)通过截断KOA冷痛敏扭转其病情进展的疗效机制。方法:1.对比KOA的人滑膜组织(全膝关节置换手术术中提取废弃滑膜)和非KOA的人滑膜组织(半月板损伤关节镜清理修补手术中所获滑膜碎屑)中TRPA1和TRPM8的蛋白和mRNA含量;2.对比KOA大鼠模型冷刺激痛阈、滑膜组织TRPA1和TRPM8的蛋白和mRNA含量变化,以及选择性TRPA1、TRPM8阻断剂对冷痛阈的影响和滑膜细胞TRPA1、TRPM8电生理特性的改变。3.观察"易层"贴敷膏剂对KOA大鼠模型冷刺激痛阈、滑膜组织TRPA1和TRPM8蛋白和mRNA含量的影响。4.TRPA1和TRPM8活化的维持尚受到炎性细胞因子的调节,进一步观察"易层"贴敷膏剂对KOA炎性细胞因子含量的影响。结果:1.KOA的人滑膜组织对比非KOA的人滑膜组织,TRPA1、TRPM8的蛋白和mRNA含量水平显著上调,差异有统计学意义(P0.05);2.KOA大鼠对比空白对照组健康大鼠冷刺激痛阈显著下降,TRPA1、TRPM8的蛋白和mRNA含量水平显著上调,差异有统计学意义(P0.05);选择性TRPA1、TRPM8阻断剂通过阻断滑膜细胞上TRPA1、TRPM8的钙离子内流,能够上调KOA大鼠的冷刺激痛阈,差异有统计学意义(P0.05);3."易层"贴敷膏剂能够上调KOA大鼠模型冷刺激痛阈,下调KOA大鼠滑膜组织中TRPA1和TRPM8蛋白和mRNA含量,差异有统计学意义(P0.05);4."易层"贴敷膏剂能够下调KOA大鼠促炎细胞因子含量水平,上调抑炎细胞因子含量水平,这与其能够抑制TRPA1、TRPM8的通道活化有关。结论:TRPA1和TRPM8是KOA冷刺激痛敏的重要介导者,"易层"贴敷膏剂可能通过下调TRPA1、TRPM8表达和抑制TRPA1、TRPM8通道活性,缓解KOA冷刺激痛敏,发挥其疗效。"温经活血、截断扭转"作为KOA治疗理论有其临床价值。
[Abstract]:Knee osteoarthritis (Knee Osteoarthritis,KOA) is the most common degenerative disease of bone and joint, because its pathogenesis has not been clear, so far there is no special therapeutic drug. KOA pain symptoms appeared very early. KOA pain is also the most far-reaching. KOA pain is very sensitive to cold stimulation. In recent years, studies have gradually revealed that this is a kind of cold stimulation pain sensitivity. TRPA1 and TRPM8 are the mediators of pain signal transmission caused by hypothermia stimulation in human body, and whether there is TRPA1, in knee osteoarthritis. The involvement of TRPM8 is important for understanding KOA pain and developing targeted drugs. " Yi layer "paste is a patented prescription in our hospital. With the legislation of" warming meridians and activating blood circulation ", it has a good effect by" cutting off "KOA pain and" reversing "the progress of KOA. Whether its efficacy mechanism is related to this. It is also the focus of our research. Objective: to explore the relationship between TRPA1 and TRPM8 and KOA cold stimulation hyperalgesia, and the therapeutic mechanism of warming meridians and activating blood circulation to reverse the progress of KOA cold hyperalgesia by cutting off KOA cold hyperalgesia. Methods: 1. The protein and mRNA contents of TRPA1 and TRPM8 in human synovial tissue of KOA (discarded synovium extracted during total knee arthroplasty) and non-KOA synovial tissue (synovial debris obtained during meniscus injury arthroscopic repair) were compared. 2. The changes of protein and mRNA content of TRPA1 and TRPM8 in synovial tissue, the effect of selective TRPA1,TRPM8 blocker on cold pain threshold and the changes of TRPA1,TRPM8 electrophysiological characteristics of synovial cells were compared with those of cold stimulation pain threshold, synovial tissue protein and mRNA content. To observe the effect of "Yi layer" paste on the pain threshold of cold stimulation and the content of TRPA1 and TRPM8 protein and mRNA in synovial tissue of KOA rats. 4. The maintenance of TRPA1 and TRPM8 activation is still regulated by inflammatory cytokines. Further observe the effect of "Yi layer" paste on the content of inflammatory cytokines in KOA. Results: the content of TRPA1,TRPM8 protein and mRNA in human synovial tissue of 1.KOA was significantly higher than that of non-KOA synovial tissue, and the difference was statistically significant (P 0.05). Compared with the blank control group, the cold stimulation pain threshold of 2.KOA rats was significantly decreased, and the levels of TRPA1,TRPM8 protein and mRNA were significantly up-regulated (P 0.05). Selective TRPA1,TRPM8 blockers can up-regulate the cold stimulation pain threshold of KOA rats by blocking the calcium influx of TRPA1,TRPM8 on synovial cells, the difference is statistically significant (P 0.05). " Yi layer "paste can up-regulate the pain threshold of cold stimulation in KOA rat model and down-regulate the contents of TRPA1 and TRPM8 protein and mRNA in synovial tissue of KOA rats, the difference is statistically significant (P 0.05)." Yi layer "paste can down-regulate the content of pro-inflammatory cytokines and up-regulate the level of anti-inflammatory cytokines in KOA rats, which is related to the inhibition of TRPA1,TRPM8 channel activation. Conclusion: TRPA1 and TRPM8 are important mediators of KOA cold stimulation hyperalgesia. "Yi layer" paste may relieve KOA cold stimulation pain sensitivity and exert its curative effect by down-regulating the expression of TRPA1,TRPM8 and inhibiting the activity of TRPA1,TRPM8 channel. " Wenjing Huoxue, truncated torsion "as the treatment of KOA theory has its clinical value."
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R274.9
本文编号：2472975
[Abstract]:Knee osteoarthritis (Knee Osteoarthritis,KOA) is the most common degenerative disease of bone and joint, because its pathogenesis has not been clear, so far there is no special therapeutic drug. KOA pain symptoms appeared very early. KOA pain is also the most far-reaching. KOA pain is very sensitive to cold stimulation. In recent years, studies have gradually revealed that this is a kind of cold stimulation pain sensitivity. TRPA1 and TRPM8 are the mediators of pain signal transmission caused by hypothermia stimulation in human body, and whether there is TRPA1, in knee osteoarthritis. The involvement of TRPM8 is important for understanding KOA pain and developing targeted drugs. " Yi layer "paste is a patented prescription in our hospital. With the legislation of" warming meridians and activating blood circulation ", it has a good effect by" cutting off "KOA pain and" reversing "the progress of KOA. Whether its efficacy mechanism is related to this. It is also the focus of our research. Objective: to explore the relationship between TRPA1 and TRPM8 and KOA cold stimulation hyperalgesia, and the therapeutic mechanism of warming meridians and activating blood circulation to reverse the progress of KOA cold hyperalgesia by cutting off KOA cold hyperalgesia. Methods: 1. The protein and mRNA contents of TRPA1 and TRPM8 in human synovial tissue of KOA (discarded synovium extracted during total knee arthroplasty) and non-KOA synovial tissue (synovial debris obtained during meniscus injury arthroscopic repair) were compared. 2. The changes of protein and mRNA content of TRPA1 and TRPM8 in synovial tissue, the effect of selective TRPA1,TRPM8 blocker on cold pain threshold and the changes of TRPA1,TRPM8 electrophysiological characteristics of synovial cells were compared with those of cold stimulation pain threshold, synovial tissue protein and mRNA content. To observe the effect of "Yi layer" paste on the pain threshold of cold stimulation and the content of TRPA1 and TRPM8 protein and mRNA in synovial tissue of KOA rats. 4. The maintenance of TRPA1 and TRPM8 activation is still regulated by inflammatory cytokines. Further observe the effect of "Yi layer" paste on the content of inflammatory cytokines in KOA. Results: the content of TRPA1,TRPM8 protein and mRNA in human synovial tissue of 1.KOA was significantly higher than that of non-KOA synovial tissue, and the difference was statistically significant (P 0.05). Compared with the blank control group, the cold stimulation pain threshold of 2.KOA rats was significantly decreased, and the levels of TRPA1,TRPM8 protein and mRNA were significantly up-regulated (P 0.05). Selective TRPA1,TRPM8 blockers can up-regulate the cold stimulation pain threshold of KOA rats by blocking the calcium influx of TRPA1,TRPM8 on synovial cells, the difference is statistically significant (P 0.05). " Yi layer "paste can up-regulate the pain threshold of cold stimulation in KOA rat model and down-regulate the contents of TRPA1 and TRPM8 protein and mRNA in synovial tissue of KOA rats, the difference is statistically significant (P 0.05)." Yi layer "paste can down-regulate the content of pro-inflammatory cytokines and up-regulate the level of anti-inflammatory cytokines in KOA rats, which is related to the inhibition of TRPA1,TRPM8 channel activation. Conclusion: TRPA1 and TRPM8 are important mediators of KOA cold stimulation hyperalgesia. "Yi layer" paste may relieve KOA cold stimulation pain sensitivity and exert its curative effect by down-regulating the expression of TRPA1,TRPM8 and inhibiting the activity of TRPA1,TRPM8 channel. " Wenjing Huoxue, truncated torsion "as the treatment of KOA theory has its clinical value."
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R274.9
【参考文献】
相关期刊论文 前10条
1 Haris S Vasiliadis;Konstantinos Tsikopoulos;;Glucosamine and chondroitin for the treatment of osteoarthritis[J];World Journal of Orthopedics;2017年01期
2 王培民;邢润麟;;以痛为枢、截断扭转诊治膝骨关节炎理论研究[J];南京中医药大学学报;2016年06期
3 张康乐;郭艳幸;郭珈宜;李峰;张云飞;;活血止痛方治疗膝骨关节炎疼痛疗效观察[J];亚太传统医药;2016年17期
4 徐波;张来根;张农山;茆军;范竞;王培民;;“MEMS微针促透‘易层’贴敷技术”治疗膝骨关节炎372例临床研究[J];时珍国医国药;2016年08期
5 王强;陈元川;庞坚;陈博;王辉昊;曹月龙;石瑛;詹红生;石印玉;;膝骨关节炎证候分型与疼痛程度的相关性分析[J];中国中医骨伤科杂志;2016年05期
6 邢润麟;茆军;王培民;张农山;;国外局部外用药控制膝骨关节炎疼痛的临床研究进展[J];中国疼痛医学杂志;2016年02期
7 方金苗;杜武勋;;中药四气、五味药性物质基础研究[J];辽宁中医药大学学报;2015年12期
8 陈卫衡;;探索建立系统的膝骨关节炎中医临床科研范式和理论体系[J];中医正骨;2015年07期
9 唐宗湘;;温度敏感的TRP通道与中药药性四气理论[J];南京中医药大学学报;2015年04期
10 王桂珍;刘健;黄传兵;汪元;曹云祥;陈瑞莲;范海霞;葛瑶;纵瑞凯;万磊;;中药内外合治湿热痹阻型膝骨关节炎临床疗效观察[J];安徽医药;2015年08期
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