基于1HNMR代谢组学技术研究针刺经穴与非经穴治疗溃疡性结肠炎的脑代谢物质基础

Brain metabolism material basis of needling meridian acupoints and non-meridian non-acupoints for treating ulcerative colitis： a study based on 1HNMR metabonomics

目的 以溃疡性结肠炎（UC）大鼠为研究载体,运用1 HNMR代谢组学技术比较针刺足阳明经穴与非经非穴治疗UC大鼠后大脑皮层代谢物的变化及差异,从整体代谢的角度为阐释“经穴—脑相关”的脑代谢物质基础提供客观的实验依据.方法 采用葡聚糖硫酸钠（DSS）法复制UC大鼠模型,随机分为模型组、经穴组、非经非穴组,每组13只,另设相同数量的大鼠为对照.经穴组采用上巨虚、天枢和足三里进行电针治疗,非经非穴组在穴位旁开5 mm进行电针治疗.治疗结束后取直结肠和回盲部行病理形态学观察;分离脑皮层,分别提取水溶性和脂溶性脑组织代谢物,运用1 HNMR技术获取代谢谱图,将波峰归一化积分后的数值导入SIMCA-P10.0软件包,采用主成分分析和正交信号校正相结合的方法对数据进行分析,得出相应的得分图和因子载荷图.结果 DSS诱导的UC模型大鼠肠道出现典型的UC症状;针刺足三里、天枢、上巨虚对溃疡性结肠炎大鼠脑皮层代谢物有明显影响,特别是对水溶性脑代谢物的影响更明显,对脂溶性代谢产物影响较小.其中,乳酸、磷酸肌酸、N-乙酰天冬氨酰谷氨酸、低密度脂蛋白、极低密度脂蛋白等代谢物是针刺经穴作用的关键代谢物;而针刺非经非穴对溃疡性结肠炎大鼠脑皮层代谢物的影响主要表现在γ-氨基丁酸、缬氨酸等.这表明针刺非经非穴的脑代谢物与经穴组存在着差异.结论 针灸经穴和非经非穴对溃疡性结肠炎动物模型的脑代谢物有不同影响,其中脑皮层水溶性提取物的差别更为显著,脂溶性代谢产物差别不大,提示采用经穴针刺治疗溃疡性结肠炎具有一定的脑特异性代谢物质基础.

Objective To compare the changes and difference in cortex metabolites in the rats with ulcerative colitis （UC） before and after treated with needling in acupoints in Zuyangming Meridian or needling in non-meridian non-acupoints by applying 1HNMR metabonomics technique,and provide experimental evidence for explaining brain metabolism material basis of acupoint-brain correlation from the aspect of overall metabolism.Methods The rat model of UC was established by using dextran sulfate sodium （DSS） method,and the rats were randomly divided into model group,meridian acupoint group and non-meridian non-acupoint group （each n =13）.Other 13 rats were included into control group.The meridian acupoint group was treated with electro-acupuncture （EA） in Shangjuxu （ST37）,Tianshu （ST25） and Zusanli （ST36）,and non-meridian non-acupoint group was treated with EA in the points aside acupoints （5 mm）.After treatment,colorectal and ileocecal tissues were collected for observing pathomorphological changes.The cerebral cortex tissue was isolated for extracting water-soluble and lipidsoluble brain tissue metabolites.The metabolic spectra were obtained by using 1H NMR metabonomics technique,and data was input into SIMCA-P10.0 software package after wave-peak normalization.The data was analyzed by applying principal component analysis combining orthogonal signal correction for obtaining score plots and factor loading plots.Results EA in ST36,ST25 and ST37 had significant influences on brain tissue metabolites especially on water-soluble ones in UC rats induced by DSS.The key metabolites effected by EA were lactate,creatine,N-acetyl aspartic glutamic acid,low density lipoprotein （LDL） and very low density lipoprotein （VLDL）,while EA in non-meridian non-acupoints mainly influenced γ-aminobutyric acid and valine.The results indicated that there was difference in brain tissue metabolites between EA in non-meridian non-acupoints and EA in meridian acupoints.Conclusion EA in meridian acupoints and non-meridian non-acupoints have different effects on brain tissue metabolites in UC rats,and the difference in water-soluble metabolites is more significant,and that in lipid-soluble metabolites is not significant.The results indicate that treatment of UC with EA has some brain metabolism material basis.