热补针法对寒证类风湿关节炎模型家兔滑膜组织代谢物谱的影响

摘要

Objective To compare the influence of metabolites in synovial tissues of rheumatoid arthritis ( RA) cold syndrome rab-bits treated with different needling and investigate the specificity mechanisms of heat-reinforcing needling for RA. Methods A total of 40 healthy purple blue rabbits were randomly divided into normal control ( NC ) , model control ( MC ) , reinforcing-reducing needling group (RRN),twirling-reinforcing needling group (TRN) and heat-reinforcing needling group (HRN) (n=8). RA cold syndrome rabbit model was made with ovalbumin and freezing. Except NC and MC,RRN was given acupuncture of reinforcing-reducing needling at Zusanli (ST36), TRN was given acupuncture of twirling-reinforcing needling at Zusanli (ST36),and HRN was administrated acupuncture of heat-reinforcing needling at Zusanli (ST36),once a day and retaining 30 min,for 7 times. Fresh synovial tissues of rabbit knee joints were extracted after the intervention,then liquid chromatography quadrupole time-of-flight mass spectrometry ( LC-Q/TOF-MS) technology was used to evaluate meta-bolic profiles. All the data were analyzed by principal component analysis ( PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) by using the SIMCA-P(13. 0) software for windows. The results were compared. Results The synovial tissues metabolites concerning alanine,citrulline,proline,glutamine,glycine,glutamic acid,sphingolin1-phosphoric acid,lysoph-osphatidylcholine,creatine,iso-leucine,leucine,and valine in MC mainly were decreased compared with NC(P<0. 05). The above-mentioned metabolites were increased compared with MC and all acupuncture groups (P<0. 05),but the metabolites concerning creatine,isoleucine,leucine,valine in HRN were obviously increased than those of RRN and TRN (P<0. 05). Conclusions By adjusting the amino acids metabolism which is associated with RA energy metabolism,heat-reinforcing needling has specific effect on RA by improving supply of amino acids and relieving negative ni-trogen balance.%目的 探讨比较不同针法对寒证类风湿关节炎(RA)模型家兔滑膜组织代谢物谱的影响及热补针法治疗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.以卵蛋白诱导联合低温冷冻法建立寒证RA模型.除正常组、模型组外,其余各组分别于足三里穴施以平补平泻、捻转补法、热补针法针刺,1次/d,留针30 min,共7次.治疗结束后分离家兔膝关节滑膜组织,采用液相色谱-四级杆飞行时间质谱(LC-Q/TOF-MS)检测滑膜组织代谢物,利用主成分分析(PCA)和正交偏最小二乘方判别分析(OPLS-DA)方法对数据进行统计分析.结果 与正常组比较,模型组滑膜组织代谢物变化主要体现于丙氨酸、瓜氨酸、脯氨酸、谷氨酰胺、甘氨酸、谷氨酸、鞘氨醇1-磷酸、溶血卵磷脂、肌酸、异亮氨酸/亮氨酸、缬氨酸含量降低(P<0.05);与模型组比较,各针刺组上述代谢物含量增高(P<0.05),且热补针法组肌酸、异亮氨酸/亮氨酸、缬氨酸含量明显高于平补平泻组和捻转补法组.结论 热补针法通过调节与RA能量代谢相关的氨基酸代谢,在改善RA机体氨基酸的供给及减缓负氮平衡方面有一定的特异性作用.