热补针法干预风湿性关节炎寒证模型家兔尿液代谢组学研究?

袁博++王金海++张星华++田亮++任朝展++杜小正

摘要：目的 通过对风湿性关节炎（RA）寒证模型家兔尿液中内源性代谢物的分析，探讨热补针法治疗RA的特异性机制。方法 采用卵蛋白诱导联合低温冷冻建立RA寒证家兔模型。将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。分别应用平补平泻、捻转补法、热补针法针刺“足三里”，1次/d，留针30 min，共7 d。干预结束后收集新鲜尿液，气相色谱-四级杆飞行时间质谱采集各组尿液代谢物图谱，利用主成分分析和偏最小二乘判别分析对数据进行统计。结果 与正常组比较，模型组尿液代谢物变化显著，氨基酸代谢相关物质降低，三羧酸循环、糖类代谢、脂肪酸代谢相关物质增高；与模型组比较，各针刺组三羧酸循环、脂质代谢相关物质降低（P<0.05）；热补针法组三羧酸循环相关物质明显低于平补平泻组和捻转补法组（P<0.05）。结论 热补针法治疗RA的特异性体现在对三羧酸循环的调控。

关键词：热补针法；风湿性关节炎；代谢组学；系统生物学；气相色谱-四级杆飞行时间质谱；兔

DOI：10.3969/j.issn.1005-5304.2016.12.016

中图分类号：R245 文献标识码：A 文章编号：1005-5304（2016）12-0064-05

Study on Intervention of Heat-reinforcing Needling for Metabonomics of Rabbits Urine with Rheumatoid Arthritis Cold Syndrome YUAN Bo1， WANG Jin-hai2， ZHANG Xing-hua1， TIAN Liang1， REN Chao-zhan3， DU Xiao-zheng1 （1. Gansu University of Chinese Medicine， Lanzhou 730000， China； 2. The Second Hospital of Lanzhou University， Lanzhou 730030， China； 3. Gansu Provincial Hospital of Traditional Chinese Medicine， Lanzhou 730050， China）

Abstract： Objective To analyze the endogenous metabolites in urine of rabbits with rheumatoid arthritis （RA） cold syndrome； To investigate the specificity mechanisms of heat-reinforcing needling for RA. Methods Egg albumin inducing method combined with low temperature and freezing method was used to establish RA cold syndrome rabbit models. A total of 40 healthy purple blue rabbits were randomly divided into normal control group （NC）， model control （MC）， reinforcing-reducing needling group （RRN）， twirling-reinforcing needling group （TRN） and heat-reinforcing needling group （HRN） （n=8/group）. 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， and HRN was administrated acupuncture of heat-reinforcing needling at Zusanli， once a day and retaining 30 min， a total of seven days. Fresh urine sample after the intervention was collected and then gas chromatography quadrupole time-of-flight mass spectrometry technology were used to evaluate metabolic profiles. All the data were analyzed by principal component analysis and partial least squares discriminant analysis. Results Compared with NC， the urine metabolites of MC changed significantly， mainly reflecting in the increase of amino acid metabolism were decreased， metabolites of TCA cycle， carbohydrate and fatty acid metabolism were increased； Compared with MC， the urine metabolites of TCA cycle and fatty acid metabolism were decreased in all intervention of acupuncture group （P<0.05）， but the metabolites of TCA cycle in HRN were obviously less than that of RRN and TRN （P<0.05）. Conclusion The specificity of heat-reinforcing needling for RA manifests in regulation of TCA cycle.

Key words： heat-reinforcing needling； rheumatoid arthritis； metabonomics； systems biology； GC-Q/TOF-MS； rabbits

基金项目：国家自然科学基金（81260558）；甘肃省自然科学基金（1208RJZA185）；甘肃省高校基本科研业务费项目（2011年）

通讯作者：杜小正，E-mail：lz-duxiaozheng@163.com

风湿性关节炎（rheumatoid arthritis，RA）是一种自身免疫性疾病。目前尚无根治方法，针灸具有抗炎、免疫调节等作用[1-3]，对RA具有确切的临床疗效[4]。热补针法是郑魁山教授简化烧山火手法和进火法而形成的一种复式手法，具有补益正气、温阳散寒功效。课题组前期研究表明，热补针法在治疗RA过程中具有相对特异性的效果[5-8]。但前期研究仅从镇痛方面进行了初步分析，缺乏对针刺后机体产生的全部生物信息的综合分析。……