Abstract：
To explore the mechanism of the classic formula Sanpian Decoction in treating chronic migraine, this study employed the four-dimensional data-dependent acquisition(4D-DIA) proteomics to analyze the effect of the decoction on chronic migraine in rats and experimentally verified the key differentially expressed proteins. Firstly, SD male rats were randomly divided into groups and repeatedly injected with nitroglycerin to prepare a chronic migraine model. After 7 consecutive days of gavage, rat grimace scale(RGS) was employed to evaluate the treatment efficacy. The trigeminal ganglion was collected for 4D-DIA proteomics, on the basis of which the diffe-rentially expressed proteins between groups were screened. Multiple databases were used for the Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment of the differentially expressed proteins. STRING and Cytoscape were employed to establish the protein-protein interaction(PPI) network. Western blot was employed to determine the expression level of the key diffe-rentially expressed protein TRPV1. The results showed that there were 517 differentially expressed proteins between blank group and model group and 221 differentially expressed proteins between model group and medium-dose Sanpian Decoction group. The GO and KEGG enrichment results showed that these differentially expressed proteins were mainly related to inflammatory response, injurious sensory stimulation, triglyceride metabolism, immune regulation, etc., which mainly involved the inflammation-related TRP, AMPK, PI3K-Akt, and TGF-β signaling pathways. The PPI network showed that the target proteins such as IGF, TOP2A, APOA1, CDK1, TTN, RYR1, and CSRP3 had high degrees. Compared with that in model group, the expression level of TRPV1 altered in medium-and high-dose Sanpian Decoction group(P<0.05). In conclusion, Sanpian Decoction may treat chronic migraine by regulating the inflammation-related pathways such as TRP, AMPK, and PI3K-Akt. It plays an important role in the regulation of TRPV1 protein and potentially modulates the perception of injurious stimuli, lipid metabolism, and immune responses.
摘要：
探讨经典方三片汤治疗慢性偏头痛的作用机制,这项研究采用四维数据依赖采集(4D-DIA)蛋白质组学方法分析了汤剂对大鼠慢性偏头痛的影响,并通过实验验证了关键的差异表达蛋白.首先,将SD雄性大鼠随机分组,反复注射硝酸甘油制备慢性偏头痛模型。连续7天灌胃后，采用大鼠鬼脸量表（RGS）评估治疗效果。收集三叉神经节进行4D-DIA蛋白质组学,在此基础上筛选了组间差异表达的蛋白质。多个数据库用于基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集差异表达的蛋白质。STRING和Cytoscape用于建立蛋白质-蛋白质相互作用（PPI）网络。Western印迹用于确定关键差异表达蛋白TRPV1的表达水平。结果表明，空白组与模型组之间有517种差异表达蛋白，模型组与中剂量三片汤组之间有221种差异表达蛋白。GO和KEGG富集结果显示,这些差异表达蛋白主要与炎症反应有关,有害的感觉刺激，甘油三酯代谢，免疫调节，等。,主要涉及炎症相关的TRP，AMPK,PI3K-Akt,和TGF-β信号通路。PPI网络显示，IGF等靶蛋白，TOP2A,APOA1,CDK1,TTN,RYR1和CSRP3均有较高的程度。与模型组相比，中、高剂量三片汤组TRPV1表达水平发生改变(P<0.05)。总之,三片汤可能通过调节TRP等炎症相关通路治疗慢性偏头痛,AMPK,和PI3K-Akt。它在TRPV1蛋白的调节中起重要作用，并可能调节伤害性刺激的感知，脂质代谢，和免疫反应。
