摘要
目的基于HMGB1/CXCL12/CXCR4信号轴研究伤科冷痛贴对KOA小鼠冷痛敏的干预机制。方法使用碘乙酸钠(MIA)膝关节注射以构建膝骨关节炎(KOA)小鼠模型,提取培养小鼠外周血循环单核细胞并尾静脉回输,动物活体成像观察单核细胞募集部位。于不同时间点测定各组小鼠冷刺激痛敏阈值。HE染色评估滑膜病理改变水平,ELISA检测小鼠血清中炎症因子IL-1β、TNF-α和疼痛介质CGRP、Substance P表达,Western blot和qPCR检测滑膜组织中的TRPA1、TRPM8、HMGB1、CXCL12、CXCR4、CollagenⅠ、Netrin-1以及背根神经节(DRG)组织中的DCC等冷痛敏相关指标的蛋白和基因表达。结果动物活体成像显示,循环单核细胞尾静脉回输至KOA小鼠后可在膝关节部位募集,且HMGB1组小鼠循环单核细胞在膝关节部位募集更多。此外,与对照组比较,KOA组和HMGB1组小鼠较对照组小鼠滑膜出现炎性病理改变,血清炎症因子和疼痛介质表达升高,冷痛敏阈值降低,滑膜组织和DRG组织中冷痛敏指标蛋白和基因表达上调,且相较于KOA组,HMGB1组小鼠上述改变更为显著(P<0.05);使用伤科冷痛贴或GL干预后,小鼠滑膜炎症减轻,血清炎症因子和疼痛介质减少,冷痛敏阈值上调,且冷痛敏相关指标蛋白和基因表达水平的上调被显著逆转(P<0.05)。结论伤科冷痛贴可能通过抑制HMGB1/CXCL12/CXCR4信号通路改善KOA小鼠的滑膜炎症和冷痛敏症状。
OBJECTIVE To explore the intervention mechanism of Shangke Lengtongtie on cold hyperalgesia in KOA mice based on the HMGB1/CXCL12/CXCR4 signaling axis.METHODS Monosodium iodoacetate(MIA)was used for the intra-articular injection into the knee joint to establish mice model of knee osteoarthritis(KOA).Peripheral blood monocytes were extracted from mice,cultured,and then reinfused into the tail vein of the mice.Subsequently,in vivo animal imaging was used to observe the recruitment sites of these monocytes.The cold hyperalgesia threshold was measured at various time points in each group of mice.Hematoxylin and eosin(HE)staining was used to evaluate the level of synovial pathological changes.ELISA was employed to detect the expression of inflammatory factors IL-1β,TNF-α,and pain mediators CGRP and Substance P in mouse serum.Western blot and qPCR methods were used to detect the protein and gene expression of cold hyperalgesia-related indicators such as TRPA1,TRPM8,HMGB1,CXCL12,CXCR4,CollagenⅠ,and Netrin-1 in synovial tissue,as well as DCC in dorsal root ganglia(DRG)tissue.RESULTS In vivo imaging showed that after the monocytes were reinfused into KOA mice,they were recruited to the knee joint area,with the HMGB1 group exhibiting a greater recruitment of circulating monocytes at the knee joint.Additionally,compared to the control group,the KOA group and HMGB1 group showed inflammatory pathological changes in the synovium,increased expression of serum inflammatory factors and pain mediators,reduced cold hyperalgesia threshold,and upregulated protein and gene expression of cold hyperalgesia-related indicators in synovial and DRG tissues.The changes were more significant in the HMGB1 group compared to the KOA group(P<0.05).After treatment with Shangke Lengtongtie or GL intervention,synovial inflammation was alleviated,serum inflammatory factors and pain mediators decreased,cold hyperalgesia threshold increased,and the upregulation of cold hyperalgesia-related indicator protein and gene expression levels was significantly reversed(P<0.05).CONCLUSION Shangke Lengtongtie exerts a beneficial effect on the mitigation of synovitis and cold hyperalgesia in KOA mice,a therapeutic mechanism that possibly mediated through the inhibition of the HMGB1/CXCL12/CXCR4 signaling axis.
作者
魏义保
张力
廖太阳
揭立士
马振源
吴鹏
黄正泉
张立
丁亮
梅伟
邢润麟
殷松江
李晓辰
张农山
茆军
王培民
WEI Yibao;ZHANG Li;LIAO Taiyang;JIE Lishi;MA Zhenyuan;WU Peng;HUANG Zhengquan;ZHANG Li;DING Liang;MEI Wei;XING Runlin;YIN Songjiang;LI Xiaochen;ZHANG Nongshan;MAO Jun;WANG Pei-min(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Key Laboratory for Metabolic Diseases in Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Nanjing University of Chinese Medicine Clinical Research Institute for Knee Osteoarthritis,Nanjing 210029,China)
出处
《南京中医药大学学报》
2025年第2期195-202,共8页
Journal of Nanjing University of Traditional Chinese Medicine
基金
江苏省医学重点学科/实验室建设单位(JSDW202252)
江苏省中医院中医膝骨关节炎临床医学创新中心(Y2023zx05)
南京中医药大学膝骨关节炎临床专病研究院(LCZBYJYZZ2024-003)。
作者简介
第一作者:魏义保,男,博士研究生,E-mail:a398645461@163.com;通信作者:王培民,男,主任中医师,博士生导师,主要从事膝骨关节炎发病机制及中药干预机制研究,E-mail:drwpm@163.com。
