电针“梁门”-“足三里”对急性胃黏膜损伤模型大鼠血清代谢物动态表达的影响

Effects of Electroacupuncture at “Liangmen”(ST 21) and “Zusanli”(ST 36) on Dynamic Expression of Serum Metabolites in Acute Gastric Mucosal Injury Model Rats

目的采用核磁共振氢谱技术(1H NMR)探讨电针梁门-足三里治疗急性胃黏膜损伤的可能作用机制。方法 54只SD大鼠随机分为正常组、模型组、电针组,各组又分为干预1、4、7天3个时间点,每个时间点6只。除正常组外其余各组采用75%乙醇灌胃法复制急性胃黏膜损伤大鼠模型。造模成功后电针组大鼠用鼠板捆绑固定,采用疏密波(4 Hz,50 Hz),电针"梁门"(负极)"足三里"(正极),每次30 min,每日1次。正常组和模型组大鼠用鼠板捆绑固定30 min,每日1次。分别于干预1、4、7天后光镜观察胃黏膜病理形态,检测大鼠血清1H NMR谱,并利用模式识别方法分析不同干预时间下代谢物谱的差异,用正交偏最小二乘判别分析(OPLS-DA)模型和独立样本t检验,以P <0. 05筛选潜在的生物代谢标志物。结果病理结果显示,正常组大鼠胃黏膜表面细胞完整,未见溃疡性损害。干预1、4天时模型组大鼠可见胃黏膜上皮结构破坏严重,毛细血管增生及出血;干预4天时电针组大鼠胃黏膜的黏膜结构恢复较为完整,较模型组明显改善。血清代谢组学结果显示,正常组与模型组分离出差异代谢物6个:甘油磷酸胆碱、谷氨酰胺、甜菜碱、葡萄糖、乙酸盐和胆碱。与模型组比较,干预1天时电针组胆碱、甘油磷酸胆碱、葡萄糖、乙酸盐、谷氨酰胺和甜菜碱均升高;干预4天时胆碱、葡萄糖、谷氨酰胺和甜菜碱升高,甘油磷酸胆碱和乙酸盐降低;干预7天时甘油磷酸胆碱降低,葡萄糖、乙酸盐、谷氨酰胺和甜菜碱升高,胆碱无明显变化。结论电针"梁门"-"足三里"可有效回调急性胃黏膜损伤大鼠血清相关差异代谢物,恢复相应代谢通路稳定,以电针4天效果最佳,可能是其逆转胃黏膜损伤的作用机制之一。

Objective To investigate the possible mechanism of electroacupuncture at Liangmen(ST 21)and Zusanli(ST 36)in the treatment of acute gastric mucosal injury by nuclear magnetic resonance spectroscopy(1 H NMR).Methods A total of 54 SD rats were randomly divided into normal group,model group and electroacupuncture group.Each group was divided into three subgroup treated for 1,4 and 7 days respectively,with 6 rats at each subgroup.Except for the normal group,the rat models of acute gastric mucosal injury were replicated by 75%ethanol.After successful modeling,the rats in the electroacupuncture group were fixed with the rat plates,and the electroacupuncture at'Liangmen'(ST 21)(negative polar)and'Zusanli'(ST 36)(positive polar)were performed 30 mins,once a day.Dilatational wave(4 Hz,50 Hz)was used.Rats in the normal group and the model group were fixed with a rat plate for 30 min once a day.The pathological morphology of gastric mucosa was observed by light microscopy after 1,4,and 7 days of intervention,and the serum1 H NMR spectra were detected.The pattern recognition method was used to analyze the differences of metabolite profiles under different intervention time.The orthogonal partial least squares discriminant analysis(OPLS-DA)and independent sample t-test were used to screen the potential biometabolism markers at the level P<0.05.Results Pathological results showed that the surface of the gastric mucosa of the normal group was intact and no ulcerative damage was observed.After 1 and 4 days of intervention,the gastric mucosal epithelial structure was severely damaged,and capillary proliferation and hemorrhage occurred in the model group.The mucosal structure of the gastric mucosa in the electroacupuncture group was completely intact after 4 days of intervention,which was significantly improved compared with the model group.Serum metabolomics results showed that the normal group and the model group were separated from 6 metabolites:glycerol phosphate,glutamine,betaine,glucose,acetate and choline.Compared with the model group,choline,glycerophosphocholine,glucose,acetate,glutamine and betaine were increased in the electroacupuncture group after 1 day of intervention;After 4 days of intervention,choline,glucose,glutamine and glycine phosphorylcholine and acetate increased,and betaine,glycerol phosphate choline decreased;After 7 days of intervention,glycerophosphoryl choline decreased,glucose,acetate,glutamine and betaine increased,and choline did not change significantly.Conclusion Electroacupuncture at Liangmen(ST 21)and Zusanli(ST 36)can effectively alleviate the serum-related differential metabolites in rats with acute gastric mucosal injury and restore the stability of the corresponding metabolic pathway.The effect of electroacupuncture is best for 4 days,which may be one of the mechanisms of its reversal of gastric mucosal injury.