为探讨经典名方散偏汤对慢性偏头痛的作用机制，该研究采用4D-DIA蛋白质组学技术分析其对慢性偏头痛模型大鼠的干预影响，并对关键差异蛋白进行实验验证。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。连续灌胃7 d后，采用大鼠痛苦面容量表(RGS)评估药效。取三叉神经节进行4D-DIA相对定量蛋白质组学检测，筛选各组差异蛋白，结合多个数据库进行基因本体论(Gene Ontology，GO)功能和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)通路富集分析;使用STRING数据库及Cytoscape软件进行蛋白-蛋白互作(PPI)网络分析，并对关键差异表达蛋白TRPV1进行蛋白免疫印迹法(Western blot)检测。结果显示，空白组与模型组间有517个差异表达蛋白，模型组与散偏汤中剂量组间有221个差异表达蛋白。GO功能和KEGG通路富集分析显示，差异表达蛋白主要与炎症反应、伤害性感受刺激、甘油三酯代谢、免疫调节等相关，主要集中在炎症相关TRP通路、AMPK信号通路、PI3K-Akt信号通路、TGF-β信号通路等。PPI 网络显示，IGF、TOP2A、APOA1、CDK1、TTN、RYR1、CSRP3 等靶蛋白具有较高的度值(degree)。Western blot 结果显示，与模型组比较，散偏汤中、高剂量组TRPV1表达水平差异有统计学意义(P＜0。05)。综上，散偏汤可能通过调节TRP、AMPK、PI3K-Akt等炎症相关通路治疗慢性偏头痛，其对TRPV1蛋白的调节发挥了重要作用，并对伤害性刺激感受、脂代谢和免疫反应等具有潜在的调节作用。
4D-DIA proteomics reveals mechanism of Sanpian Decoction in treating chronic migraine in rats
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 em-ployed 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 Ency-clopedia 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 in-flammation-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.
Sanpian Decoctionchronic migraine4D-DIA proteomicsTRP pathwayrat model
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万方数据
