小续命汤抗缺血性脑卒中网络药理学分析及其抗炎和神经血管保护机制的体内验证

doi:10.5246/jcps.2022.05.030

中国药学(英文版) ›› 2022, Vol. 31 ›› Issue (5): 343-359.DOI: 10.5246/jcps.2022.05.030

小续命汤抗缺血性脑卒中网络药理学分析及其抗炎和神经血管保护机制的体内验证

杨滢霖¹^,², 张姗姗¹^,², 刘漫¹^,², 刘冬妮¹^,², 王月华¹^,²^,^*(), 杜冠华¹^,²^,^*()

1. 中国医学科学院药物研究所天然药物活性物质与功能国家重点实验室, 北京 100050
2. 中国医学科学院药物研究所药物靶点研究与新药筛选北京市重点实验室, 北京 100050

收稿日期:2021-12-12 修回日期:2022-02-18 接受日期:2022-03-06 出版日期:2022-06-02 发布日期:2022-06-02
通讯作者: 王月华, 杜冠华
作者简介:
+ Tel.: +86-10-63165313, E-mail: wangyuehua@pku.org.cn
+ Tel.: +86-10-63165184, E-mail: dugh@imm.ac.cn
基金资助:
National Natural Science Foundation of China (Grant No. 81473383), the Medical and Health Innovation Project of Chinese Academy of Medical Sciences (Grant No. 2016-I2M-3-007), and Innovation Fund for Graduate of Beijing Union Medical College (Grant No. 2018-1007-04).

Network pharmacological analysis of Xiao-Xu-Ming decoction against ischemic stroke and verification of its mechanism of anti-inflammation and neurovascular protection in vivo

Yinglin Yang¹^,², Shanshan Zhang¹^,², Man Liu¹^,², Dongni Liu¹^,², Yuehua Wang¹^,²^,^*(), Guanhua Du¹^,²^,^*()

1 State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
2 Beijing Key Laboratory of Drug Target Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China

Received:2021-12-12 Revised:2022-02-18 Accepted:2022-03-06 Online:2022-06-02 Published:2022-06-02
Contact: Yuehua Wang, Guanhua Du

摘要/Abstract

摘要：

中风是导致严重残疾和死亡的主要原因。中药复方小续命汤是治疗中风及其后遗症的有效方剂, 但其有效成分和作用机制尚不清楚。本研究旨在运用网络药理学方法探讨小续命汤治疗脑缺血的有效成分和作用机制。通过TCMSP数据库和分析平台数据库, 获得小续命汤12种药材的主要化学成分, 根据口服生物利用度和类药物性质筛选出小续命汤的活性成分; 通过Genecards、OMIM、TTD、Digenet和Drugbank数据库获得脑缺血疾病相关靶点; 应用Metascape数据分析平台分析小续命汤治疗脑缺血的病理生理过程和途径。结果表明, β-谷甾醇、山奈酚、槲皮素、豆甾醇、汉黄芩素和儿茶素可能是小续命汤治疗脑缺血的潜在核心活性成分, 小续命汤对中风的治疗作用与调节神经炎症反应和神经血管保护机制密切相关。在脑缺血大鼠模型上, 进一步验证了小续命汤治疗脑缺血机制与抗神经炎症和神经血管保护作用相关。综上所述, 本研究揭示了小续命汤治疗脑缺血机制与抗神经炎症及神经血管保护相关。

关键词: 小续命汤, 脑缺血, 网络药理学, 神经保护

Abstract:

Stroke is a major cause of severe disability and death. Xiao-Xu-Ming decoction (XXMD) is an effective prescription for stroke and its sequelae, while its effective ingredients and mechanism are still unclear. In the present study, we aimed to explore the effective ingredients and mechanism of XXMD in treating cerebral ischemia using network pharmacology. The main chemical components and targets of 12 herbs of?XXMD were obtained by the TCMSP database and analysis platform database. The active components in XXMD were screened according to oral utilization and drug-like properties. Then, the cerebral ischemia targets were obtained through GeneCards, OMIM, TTD, Diligent and Drugbank databases. We analyzed the pathophysiological processes and pathways involved in the treatment of cerebral ischemia with XXMD by using the Metascape data analysis platform. Results showed that β-sitosterol, kaempferol, quercetin, stigmasterol, wogonin, and catechins might be the potential core active ingredients of XXMD in the treatment of cerebral ischemia. The therapeutic effect of XXMD on stroke was mainly exerted through regulating neuroinflammatory response and neurovascular protection. Furthermore, the anti-neuroinflammation and neurovascular protection of XXMD were further confirmed using?cerebral ischemia rats. Collectively, our findings revealed that the mechanism of XXMD on the treatment of cerebral ischemia was related to anti-neuroinflammation and neurovascular protection.

Key words: Xiao-Xu-Ming decoction, Cerebral ischemia, Network pharmacology, Neuroprotection

Supporting:

杨滢霖, 张姗姗, 刘漫, 刘冬妮, 王月华, 杜冠华. 小续命汤抗缺血性脑卒中网络药理学分析及其抗炎和神经血管保护机制的体内验证[J]. 中国药学(英文版), 2022, 31(5): 343-359.

Yinglin Yang, Shanshan Zhang, Man Liu, Dongni Liu, Yuehua Wang, Guanhua Du. Network pharmacological analysis of Xiao-Xu-Ming decoction against ischemic stroke and verification of its mechanism of anti-inflammation and neurovascular protection in vivo[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(5): 343-359.

图/表 6

Figure 1. XXMD-active ingredient-targets network analysis. XXMD is composed of 12 Chinese medicinal materials. The yellow circular icons represent medicinal materials, the circular icons around each medicinal material represent the ingredients it contains, and the green diamond icons represent the genes regulated by each ingredient.

Figure 2. Venn diagram of targets related to ischemic stroke and XXMD. The purple area represents the target of cerebral ischemia, and the blue area represents the target of XXMD. There are 2324 disease targets related to cerebral ischemia. There are 284 target genes related to XXMD, and 206 common genes are identified.

Figure 3. Core target gene PPI networks analysis. (A) Workflow for core target gene PPI networks analysis; (B) PPI networks for the core target gene. The yellow circular nodes in the network represent core target genes.

Figure 4. GO and KEGG pathway analysis. (A) KEGG pathway analysis; (B) GO-BP analysis; (C) GO-CC analysis; (D) GO-MF analysis.

Figure 5. Effect of XXMD on inflammatory pathways in cerebral ischemia. (A) There are 20 pathways in the inflammatory pathways network, involving 103 gene targets. The hexagonal icons of different colors represent different signal pathways. The darker the color, the more important the pathway is. Purple diamond icons represent different genes; (B) The level of IL-1β in the cortex of rats with cerebral ischemia; (C) The level of IL-6 in the cortex of rats with cerebral ischemia (n = 10); (D) The level of TNF-α in the cortex of rats with cerebral ischemia (n = 10); (E) The level of IFN-γ in the cortex of rats with cerebral ischemia (n = 10); (F) The level of MCP-1 in the cortex of rats with cerebral ischemia (n = 10); (G) The level of ICAM-1 in the cortex of rats with cerebral ischemia (n = 10); (H) The expression of COX2 in the cortex of rats with cerebral ischemia (n = 8); (I) The expression of iNOS in the cortex of rats with cerebral ischemia (n = 8); (J) The expression of TLR4 in the cortex of rats with cerebral ischemia (n = 8); (K) The expression of MyD88 in the cortex of rats with cerebral ischemia (n = 8). Data were presented as mean ± SD. #P < 0.05, ##P < 0.01 vs. sham group; *P < 0.05, **P < 0.01 vs. IS group.

Figure 6. Effect of XXM on neurovascular-related target pathways in cerebral ischemia. (A) There are 14 neurovascular-related pathways in this network, involving 102 gene targets. The yellow octagonal icons represent different signal paths. The green diamond icons represent different genes; (B) The expression of HIF-1α in the cortex of rats with cerebral ischemia (n = 8); (C) The expression of VEGFR1 in the cortex of rats with cerebral ischemia (n = 8); (D) The expression of BDNF in the cortex of rats with cerebral ischemia (n = 8); (E) The mRNA level of BDNF in the cortex of rats with cerebral ischemia (n = 6); (F) The ratio of p-CREB/CREB in the cortex of rats with cerebral ischemia (n = 8); (G) The ratio of p-TrkB/TrkB in the cortex (n = 8). Data were presented as mean ± SD. #P < 0.05, ##P < 0.01 vs. sham group; *P < 0.05, **P < 0.01 vs. IS group.

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小续命汤抗缺血性脑卒中网络药理学分析及其抗炎和神经血管保护机制的体内验证

Network pharmacological analysis of Xiao-Xu-Ming decoction against ischemic stroke and verification of its mechanism of anti-inflammation and neurovascular protection in vivo

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