Xinjiang Institute of Materia Medica, Xinjiang Key Laboratory of Uygur Medical Research, Urumqi, Xinjiang, China.
College of traditional Chinese medicine, Xinjiang Medical University, Urumqi, Xinjiang, China.
PLoS One. 2023 Jul 26;18(7):e0289118. doi: 10.1371/journal.pone.0289118. eCollection 2023.
Cerebral ischemia-reperfusion injury (CIRI) is a major injury that seriously endangers human health and is characterized by high mortality and high disability. The total flavonoid extract of Dracocephalum moldavica L.(TFDM) in the treatment of CIRI has been proved by clinical practice. But the mechanism for the treatment of CIRI by TFDM has not been systematically revealed.
The active compounds contained in TFDM were screened by literature mining and pharmacokinetic parameters, and the targets related to CIRI were collected by searching Drugbank, Genecards and OMIM databases. Cytoscape software was used to construct the protein interaction network of TFDM for the prevention and treatment of CIRI. Geneontology and signal pathway enrichment were analyzed. The key target pathway network of TFDM compounds was constructed and verified by pharmacological experiment in vitro.
21 active components were screened, 158 potential drug targets for the prevention and treatment of CIRI were obtained, 53 main targets were further screened in the protein-protein interaction network, and 106 signal pathways, 76 biological processes, 26 cell components and 50 molecular functions were enriched (P<0.05). Through the compound-target-pathway network, the key compounds that play a role in the prevention and treatment of CIRI, such as acacetin, apigenin and other flavonoids, as well as the corresponding key targets and key signal pathways, such as AKT1, SRC and EGFR were obtained. TFDM significantly decreased LDH, MDA levels and increased the NO activity levels in CIRI. Further studies have shown that TFDM increases the number of SRC proteins, and TFDM also increases p-AKT/ AKT. Molecular docking results showed that acacetin-7-O (- 6''-acetyl) -glucopyranoside, acacetin7-O-β-D-glucopyranoside, apigenin-7-O-β-D-galactoside respectively had good affinity for SRC protein. Acacetin-7-O (- 6''-acetyl) -glucopyranoside,acacetin-7-O-β-D-glucuronide, acacetin7-O-β-D-glucopyranoside had good affinity for AKT1 protein, respectively.
Our research showed that TFDM had the characteristics of multi-component, multi-target and multi-channel in the treatment of CIRI. The potential mechanism may be associated with the following signaling pathways:1) the signaling pathways of VEGF/SRC, which promote angiogenesis, 2) the signaling pathways of PI3K/AKT, which inhibit apoptosis, and 3) acacetin-7-O (- 6''-acetyl) -glucopyranoside is expected to be used as a candidate monomer component for natural drugs for further development.
脑缺血再灌注损伤（CIRI）是严重危害人类健康的重大损伤，具有高死亡率和高致残率的特点。Dracocephalum moldavica L.（TFDM）总黄酮提取物在治疗 CIRI 方面已被临床实践所证实。但 TFDM 治疗 CIRI 的机制尚未得到系统揭示。
通过文献挖掘和药代动力学参数筛选 TFDM 中的活性化合物，通过搜索 Drugbank、Genecards 和 OMIM 数据库收集与 CIRI 相关的靶点。采用 Cytoscape 软件构建 TFDM 防治 CIRI 的蛋白质相互作用网络，进行基因本体论和信号通路富集分析。构建并验证 TFDM 化合物的关键靶标通路网络的体外药理学实验。
筛选出 21 个活性成分，获得 158 个 TFDM 防治 CIRI 的潜在药物靶点，进一步从蛋白质-蛋白质相互作用网络中筛选出 53 个主要靶点，富集到 106 条信号通路、76 个生物学过程、26 个细胞成分和 50 个分子功能（P<0.05）。通过化合物-靶标-通路网络，得到了在防治 CIRI 中起作用的关键化合物，如芹菜素、木樨草素等黄酮类化合物，以及相应的关键靶标和关键信号通路，如 AKT1、SRC 和 EGFR。TFDM 可显著降低 CIRI 模型中 LDH、MDA 水平，增加 NO 活性水平。进一步研究表明，TFDM 增加了 SRC 蛋白的数量，并且 TFDM 还增加了 p-AKT/AKT。分子对接结果表明，芹菜素-7-O-（6''-乙酰基）-葡萄糖苷、芹菜素 7-O-β-D-吡喃葡萄糖苷、木樨草素-7-O-β-D-半乳糖苷分别与 SRC 蛋白具有良好的亲和力。芹菜素-7-O-（6''-乙酰基）-葡萄糖苷、芹菜素-7-O-β-D-葡萄糖醛酸苷、芹菜素 7-O-β-D-吡喃葡萄糖苷分别与 AKT1 蛋白具有良好的亲和力。
本研究表明，TFDM 治疗 CIRI 具有多成分、多靶点、多通道的特点。其潜在机制可能与以下信号通路有关：1）VEGF/SRC 信号通路促进血管生成，2）PI3K/AKT 信号通路抑制细胞凋亡，3）芹菜素-7-O-（6''-乙酰基）-葡萄糖苷有望成为天然药物候选单体成分进一步开发。
