天麻钩藤药对治疗急性偏头痛模型大鼠的代谢组学分析

Metabolomics Analysis of Gastrodia-Uncaria in the Treatment of Acute Migraine Model Rats

目的通过代谢组学研究天麻-钩藤治疗硝酸甘油致大鼠急性偏头痛模型的作用机制。方法 40只SD大鼠随机分为正常组、模型组、琥珀酸舒马普坦组(25 mg/kg)和天麻钩藤组(4.2 g/kg),每组10只,灌胃给药7d,在第7天给药前1 h,后颈部皮下注射硝酸甘油(10 mg/kg),给药后1 h眼眶取血并收集血浆。测定血浆中一氧化氮(NO),5-羟色胺(5-HT),降钙素基因肽(CGRP)的含量,确定天麻-钩藤治疗偏头痛的药效。利用UPLC-Q-TOF-MS进行代谢组学研究,通过偏最小二乘法-判别分析(PLS-DA)进行模式识别及生物标志物的鉴定。结果与正常组比较,NO,CGRP在模型组血浆中的浓度升高,5-HT在模型组血浆中的浓度下降,且均有显著性差异(P<0.05,P<0.01),说明急性偏头痛模型建立成功;与模型组比较,给予天麻钩藤提取物后NO,CGRP浓度下降,5-HT浓度升高,均有显著性差异(P<0.05,P<0.01),证明天麻钩藤对偏头痛模型大鼠有治疗作用。通过代谢组学分析,鉴定了10个与偏头痛模型有关的生物标志物。与正常组相比,模型组中甘氨胆酸,L-精氨酸,3-吲哚硫酸盐,酪氨酸,柠檬酸含量升高;2-羟基-3-氧代己二酸,鞘氨醇-1-磷酸,L-色氨酸,亚麻酸,亚油酸含量下降。天麻钩藤给药后可使上述10个异常变化的生物标志物回调甚至恢复正常。结论天麻-钩藤主要通过调节色氨酸代谢、精氨酸代谢、亚麻酸代谢等代谢途径发挥药效。

Objective To study the mechanism of Gastrodia-Uncaria on the treatment of nitroglycerin-induced acute migraine in rats by metabolomics. Methods A total of 40 SD rats were randomly divided into normal group, model group, sumatriptan succinate group(25 mg/kg) and Gastrodia-Uncaria group(4.2 g/kg), with 10 rats in each group.Rats were intragastrically administered for 7 days. One hour before the administration on the 7 th day, nitroglycerin(10 mg/kg) was injected subcutaneously into the posterior neck, and blood was collected from the eyelids 1 hour after administration. The nitric oxide(NO), serotonin(5-HT), and calcitonin gene-related peptide(CGRP) biochemical kits were detected to determine the efficacy of Gastrodia-Uncaria in the treatment of migraine. Metabolomics studies were then performed using UPLC-Q-TOF-MS, and pattern recognition were performed by partial least squares-discriminant analysis(PLS-DA), then biomarker were identified by database alignment. Results Compared with the normal group, the concentration of NO and CGRP in the plasma of the model group increased and 5-HT in the plasma of the model group decreased. Compared with the model group, after Gastrodia-Uncaria administration, the concentrations of NO and CGRP decreased and the concentration of 5-HT increased, which proved that Gastrodia-Uncaria has therapeutic effects on migraine rats. Ten biomarkers related to the migraine model were identified. Compared with the normal group, the model group had elevated levels of glycocholic acid, L-arginine, 3-guanidine sulfate, tyrosine, and citric acid, 2-hydroxy-3-oxoadipate, the content of aminol-1-phosphate, L-tryptophan, linolenic acid and linoleic acid decreased. After administration of Gastrodia-Uncaria, the above 10 abnormally altered biomarkers could be adjusted or even returned to normal. Conclusion Gastrodia-Uncaria mainly exerts pharmacological effects by regulating metabolic pathways such as tryptophan metabolism, arginine metabolism and linolenic acid metabolism.