针刺研究

2021, v.46(02) 145-151

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基于GC-MS的针刺治疗类风湿性关节炎的代谢组学研究
Differential metabolites and metabolic pathways involving acupuncture-induced improvement of rheumatoid arthritis patients based on gas chromatography-mass spectrometry

郭耀光;孙光伟;杨玲;李聪;杨娇;
GUO Yao-guang;SUN Guang-wei;YANG Ling;LI Cong;YANG Jiao;Department of Acupuncture and Moxibustion, Affiliated Hospital of Chengdu University of TCM;Department of Oncology, Sichuan Integrative Medicine Hospital;School of Acupuncture-moxibustion and Tuina, Chengdu University of TCM;

摘要(Abstract):

目的:分析类风湿性关节炎(RA)患者血清内源性代谢物的变化,筛选差异性代谢物,观察针刺对差异性代谢物的调节作用,从代谢组学的角度探究针灸治疗RA的作用机制。方法:RA患者按随机数字表法分为针刺组及阳性药对照组,每组15例;同时收集健康体检者10例作为正常对照组。针刺组予双侧风池、阳池等穴针刺治疗,每次20 min,每天1次,治疗3个月;阳性药对照组予雷公藤多苷片口服,1片/次,每天3次,治疗3个月。观察两治疗组患者治疗前后压痛指数、肿胀指数、晨僵时间,ELISA法检测类风湿因子(RF)、C反应蛋白(CRP)含量及血红细胞沉降率(ESR)。采集正常对照组及针刺组治疗前后血清,采用GC-MS技术检测血清差异代谢物。MetaboAnalyst 4.0进行通路分析,SPSS23.0软件对差异性代谢物与药效指标进行Pearson相关性分析。结果:与治疗前比较,治疗后两治疗组压痛指数、肿胀指数、晨僵时间及RF、CRP均显著降低(P<0.05)。与正常对照组比较,共筛选出14种差异性代谢物,升高的有乳酸、柠檬酸盐、肌酸、3-羟基丁酸、花生四烯酸、缬氨酸、软脂酸;下降的有甘氨酸、色氨酸、精氨酸、L-苯丙氨酸、葡萄糖、谷氨酰胺、天冬氨酸。OPLS多变量模式分析结果显示,针刺治疗后血清差异性代谢物有向正常对照组回归的趋势。主要涉及7条代谢通路:丙氨酸、天冬氨酸和谷氨酸代谢;苯丙氨酸、酪氨酸和色氨酸代谢;甘氨酸、丝氨酸和苏氨酸代谢;乙醛酸二羧酸代谢;淀粉和蔗糖代谢;苯丙氨酸代谢;花生四烯酸代谢。柠檬酸盐、肌酸、花生四烯酸、缬氨酸、葡萄糖与压痛指数、肿胀指数、晨僵时间、RF、CRP、ESR呈正相关;甘氨酸、L-苯丙氨酸、谷氨酰胺与疗效指标呈负相关。结论:RA患者存在氨基酸及糖代谢紊乱,纠正氨基酸代谢紊乱及糖代谢可能是针灸治疗RA的作用机制之一。
Objective To analyze the endogenous metabolic biomarkers and pathways in serum involving acupuncture-induced improvement of symptoms of patients with rheumatoid arthritis(RA) by using metabolomics technique.Methods A total of 30 RA patients who were treated in the Department of Acupuncture and Moxibustion of the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine(from January 2018 to October 2018) were selected in the present study. They were randomly and equally divided into acupuncture group and medication group. Acupuncture(lifting-thrusting reinforcing and uniform reducing-reinforcing methods) was applied to bilateral Fengchi(GB20), Yangchi(TE4), Quchi(LI11), Yingu(KI10), Taixi(KI3), Xuehai(SP10), Guanyuan(CV4), Yanglingquan(GB34) and Ashi-points for 20 min every time, once daily for 3 months. Patients of the medication group were asked to take Tripterygium Wilfordii Polyglycoside tablets(a positive drug for RA, one tablet per time, 3 times a day) for 3 months. Other 10 healthy volunteers were selected as the normal control group. The tenderness scale(0-4 points) and swelling scale(0-3 points) and morning stiffness time were recorded, and serum rheumatoid factor(RF), C-reactive protein(CRP) and erythrocyte sedimentation rate(ESR) were detected for analyzing pharmacodynamic effects. Serum samples were captured for profiling and quantifying metabolite biomarkers by using GC-MS(gas chromatography-mass spectrometry) technique. The acquired metabolite profiles were processed(multivariable data analysis) by using ProteoWizard package, XCMS Online software and SIMCA 13.0 software, respectively, followed by screening differential metabolites according to variable importance projection(VIP) and by constructing metabolic pathways with MetaboAnalyst 4.0.Results After acupuncture treatment, the tenderness score, swelling score, morning stiffness time, serum RF and CRP contents were significantly decreased in both acupuncture and medication groups in comparison with their own pretreatment(P<0.05). No significant differences were found between acupuncture and medication groups in the levels of tenderness score, swelling score, morning stiffness time, serum RF and CRP contents(P>0.05). A total of 14 differential metabolites including citrate, creatine, 3-hydroxybutyrate arachidonic acid, arachidonic acid, valine, lactic acid and palmitic acid(up-regulated), and tryptophan, arginine, L-phenylalanine, glucose, glycine, glutamine, aspartic acid and(down-regulated) which mainly involve metabolic pathways of alanine, aspartic acid and glutamate; metabolism of phenylalanine, tyrosine and tryptophan; metabolism of glycine, serine and threonine; glyoxalic acid dicarboxylic acid metabolism; metabolism of starch and sucrose; and metabolism of phenylalanine and arachidonic acid, respectively.Citrate, creatine, arachidonic acid, valine and glucose was positively correlated with tenderness index, swelling index, morning stiff time, RF, CRF and ESR. Glycine, L-phenylalanine, glutamine was negatively correlated with efficacy indicators.Conclusion Acupuncture can relieve symptoms of patients with RA, which may be related to its effects in improving amino acid metabolism and glucose metabolism.

关键词(KeyWords): 类风湿性关节炎;针刺;代谢组学;GC-MS;氨基酸代谢;糖代谢
Rheumatoid arthritis;Acupuncture;Metabonomics;GC-MS;Amino acid metabolism;Glycometabolism

Abstract:

Keywords:

基金项目(Foundation): 四川省高校人文社会科学重点研究基地科研项目(No.ZYYWH1812)

作者(Author): 郭耀光;孙光伟;杨玲;李聪;杨娇;
GUO Yao-guang;SUN Guang-wei;YANG Ling;LI Cong;YANG Jiao;Department of Acupuncture and Moxibustion, Affiliated Hospital of Chengdu University of TCM;Department of Oncology, Sichuan Integrative Medicine Hospital;School of Acupuncture-moxibustion and Tuina, Chengdu University of TCM;

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DOI: 10.13702/j.1000-0607.200181

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