“热补针法”对类风湿关节炎模型家兔尿液代谢物的影响

Effects of heat-reinforcing acupuncture on urine metabolites in rheumatoid arthritis rabbits

目的:比较不同针法对类风湿关节炎(rheumatoid arthritis,RA)模型家兔尿液代谢物的影响,探讨"热补针法"治疗RA的特异性机制。方法:40只清洁级青紫蓝家兔随机分为正常组、模型组、平补平泻组、捻转补法组、热补针法组,每组8只。除正常组外,余组以卵蛋白诱导法和低温冷冻法复制RA寒证模型。平补平泻组、捻转补法组、热补针法组于"足三里"分别施以平补平泻、捻转补法、"热补针法"针刺,留针30min,每日1次,共7d。干预结束后收集24h尿液,处死家兔,剥取各组家兔膝关节滑膜组织进行组织形态学观察,采用液相色谱-质谱联用仪技术(LC-Q/TOF-MS)检测尿液代谢物,利用主成分分析(PCA)和偏最小二乘方判别分析(PLS-DA)对数据进行统计分析。结果:与正常组比较,模型组尿液代谢物变化主要体现在亮氨酸代谢相关物质降低(P<0.05),嘌呤代谢、色氨酸代谢相关物质增高(P<0.05)。与模型组比较,各针刺组尿液代谢物变化主要体现在亮氨酸代谢相关物质增高(P<0.05),色氨酸代谢、嘌呤代谢相关物质降低(P<0.05),且热补针法组亮氨酸代谢相关物质明显高于平补平泻组和捻转补法组(P<0.05)。结论:平补平泻、捻转补法及"热补针法"针刺可调控RA家兔体内亮氨酸代谢(能量代谢)、嘌呤代谢(氧化损伤)、色氨酸代谢(免疫调控)通路,而热补针法治疗RA的特异性可能体现在对亮氨酸代谢(能量代谢)的调控方面。

Objective To compare the effects of different acupuncture methods on urine metabolites in rheumatoid arthritis（RA）rabbits,and to explore the specificity mechanism of heat-reinforcing acupuncture for RA.Methods A total of 40 clean purple-blue rabbits were randomly allocated to a normal group,a model group,a mild reinforcing-reducing needling（MRRN）group,a twirling-reinforcing needling（TRN）group and a heat-reinforcing needling（HRN）group,8 rabbits in each one.Except the normal group,the rabbits in the remaining groups were treated with ovalbumin and freezing to establish RA model.The rabbits in the MRRN group,TRN group and HRN group were treated with MRRN,TRN and HRN at＂Zusanli＂（ST 36）,respectively,30 min per treatment,once a day for seven days.After treatment,24-h urine was collected.The rabbits were sacrificed to collect synovial tissues of knee to perform morphology observation;the liquid chromatography quadrupole time-offlight mass spectrometry（LC-Q/TOF-MS）was applied to measure urine metabolites.All the data were analyzed by principal component analysis（PCA）and partial least squares discriminant analysis（PLS-DA）.Results Compared with the normal group,the leucine-related metabolites,as main urine metabolites,were decreased in the model group（P〈0.05）,while the purine-related metabolites and tryptophane-related metabolites were increased（P〈0.05）.Compared with the model group,the leucine-related metabolites,as main urine metabolites,were increased in the three needling groups after treatment（P〈0.05）,while the tryptophan-related metabolites and purine-related metabolites were decreased（P〈0.05）,moreover,the leucine-related metabolites in the HRN group were obviously higher than those in the MRRN group and TRN gruop（P〈0.05）.Conclusion MRRN,TRN and HRN can regulate the pathway of leucine metabolism（energy metabolism）,purine metabolism（oxidative damage）and tryptophane metabolism（immune regulation）for RA,The specificity of HRN for RA focuses on regulation of leucine metabolism（energy metabolism）.