基于~1H-NMR代谢组学的还贝止咳方对咳嗽变异性哮喘豚鼠的干预作用

Effects of Huanbei Zhike Recipe on cough variant asthma model guinea pig based on ~1H-NMR metabolomics

目的采用~1H-NMR代谢组学方法,探讨还贝止咳方对鸡清白蛋白(OVA)致豚鼠咳嗽变异性哮喘(CVA)模型的作用机制。方法采用ip 4%OVA和ip 2%Al(OH)3制备豚鼠CVA模型,观察还贝止咳方(5、10、20 g/kg)对CVA豚鼠辣椒素引咳后的影响。给药结束后,取豚鼠右侧中叶肺组织进行~1H-NMR检测,采用多元统计分析方法探讨该方对CVA的干预作用。结果还贝止咳方能显著提高CVA豚鼠的咳嗽潜伏期,降低其咳嗽次数(P<0.01),并能明显减少肺灌洗液中的炎症因子(P<0.05)。与对照组相比,模型组豚鼠肺组织的亮氨酸、异亮氨酸、缬氨酸、丙氨酸、α-葡萄糖、乙酸、谷氨酸、甲硫氨酸、丙酮酸、琥珀酸、3-羟基丁酸酯、氧化谷胱甘肽、肌酸、乙醇胺、胆酸、甘氨酸、甘油磷酰胆碱、谷氨酰胺、甜菜碱、磷酸乙醇胺、β-葡萄糖、精氨酸水平升高,而乳酸、磷酸甲基酯的水平降低,给予还贝止咳方后能不同程度回调这24个潜在的差异代谢物。结论还贝止咳方可能通过调控能量代谢、氨基酸代谢等途径来治疗CVA。

Objective To investigate the mechanism of action of Huanbei Zhike Recipe on the cough variant asthma model（CVA） of guinea pigs by 1 H-NMR metabolomics methods. Methods The CVA model was induced by ip 4% OVA and ip 2% Al（OH）3 in guinea pigs abdominal cavity, and the effects of Huanbei Zhike Recipe（5, 10, and 20 g/kg） on the cough induced by capsaicine was investigated. After the administration, the lung of right middle lobe in guinea pigs were taken out and determinated by 1 H-NMR, and multivariate statistical analysis was used to investigate the intervention effects of Huanbei Zhike Recipe on CVA. Results The cough latency of guinea pigs can be significantly prolonged and the frequency of cough were reduced（P〈 0.01）, and the inflammatory factors in lung lavage fluid were obviously reduced（P〈 0.05）. Compared with the blank group, the levels of leucine, isoleucine, valine, alanine, α-glucose, acetic acid, glutamic acid, methionine, pyruvic acid, succinic acid, 3-hydroxybutyrate（3-HB）, GSSG, creatine, ethanolamine, cholic acid, glycine, glycerophosphoryl choline（GPC）, glutamine, betaine, phosphorylethanolamine（PE）, β-glucose, and arginine were increased, while the levels of lactate and methyl-phosphate were decreased. These 24 potential metabolites could be reversely regulated in different degree after treatment with Huanbei Zhike Recipe. Conclusion Huanbei Zhike Recipe could prevent CVA diseases likely through the regulation of energy metabolism and amino acid metabolism.