Wang Lei, Jia Jing-Nan, Wang Yi-Lin, Li Yi-le, Zou Qian-Qian, Zhang Shuo, Cui Ying-Lin
the Second Clinical Medical College, Henan University of Chinese Medicine Zhengzhou 450002, China.
the Second Clinical Medical College, Henan University of Chinese Medicine Zhengzhou 450002, China Henan Province Hospital of Traditional Chinese Medicine Zhengzhou 450002, China.
Zhongguo Zhong Yao Za Zhi. 2024 Jul;49(13):3644-3656. doi: 10.19540/j.cnki.cjcmm.20240401.501.
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.
为探讨经典方剂三偏汤治疗慢性偏头痛的作用机制，本研究采用四维数据依赖采集（4D-DIA）蛋白质组学技术分析该方对大鼠慢性偏头痛的影响，并对关键差异表达蛋白进行实验验证。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天后，采用大鼠面部表情评分（RGS）评估治疗效果。采集三叉神经节进行4D-DIA蛋白质组学分析，在此基础上筛选组间差异表达蛋白。利用多个数据库对差异表达蛋白进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析。采用STRING和Cytoscape构建蛋白质-蛋白质相互作用（PPI）网络。通过蛋白质免疫印迹法检测关键差异表达蛋白TRPV1的表达水平。结果显示，空白组与模型组之间有517个差异表达蛋白，模型组与中剂量三偏汤组之间有221个差异表达蛋白。GO和KEGG富集结果表明，这些差异表达蛋白主要与炎症反应、伤害性感觉刺激、甘油三酯代谢、免疫调节等有关，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、TOP2A、APOA1、CDK1、TTN、RYR1和CSRP3等靶蛋白具有较高的连接度。与模型组相比，中、高剂量三偏汤组TRPV1的表达水平发生改变（P<0.05）。综上所述，三偏汤可能通过调节TRP、AMPK和PI3K-Akt等炎症相关通路治疗慢性偏头痛。其在TRPV1蛋白的调节中起重要作用，并可能调节对伤害性刺激的感知、脂质代谢和免疫反应。
