Key Laboratory of Infectious Immune and Antibody Engineering of Guizhou Province, Engineering Research Center of Cellular Immunotherapy of Guizhou Province, Guizhou Medical University, Guiyang, 550025, China; School of Pharmaceutical Sciences, Guizhou Medical University, University Town, Guian New District, Guizhou, 550025, China; Immune Cells and Antibody Engineering Research Center of Guizhou Province, Key Laboratory of Biology and Medical Engineering, Guizhou Medical University, Guiyang, 550025, China; State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University, Guiyang, 550025, China.
Key Laboratory of Infectious Immune and Antibody Engineering of Guizhou Province, Engineering Research Center of Cellular Immunotherapy of Guizhou Province, Guizhou Medical University, Guiyang, 550025, China; School of Pharmaceutical Sciences, Guizhou Medical University, University Town, Guian New District, Guizhou, 550025, China; Immune Cells and Antibody Engineering Research Center of Guizhou Province, Key Laboratory of Biology and Medical Engineering, Guizhou Medical University, Guiyang, 550025, China; State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Medical University, Guiyang, 550025, China.
J Ethnopharmacol. 2023 Jan 30;301:115799. doi: 10.1016/j.jep.2022.115799. Epub 2022 Oct 8.
Sophora flavescens is a frequently used traditional Chinese medicine (TCM) for the treatment of skin disorders, diarrhea, vaginal itching and inflammatory diseases. In particular, the root of S. flavescens combination with other herbs mainly treat eczema ailment in the clinical applications. However, a holistic network pharmacology approach to understanding the mechanism by which alkaloids in S. flavescens treat eczema has not been pursued.
To examine the network pharmacological potential effect of S. flavescens on eczema, we studied the alkaloids, performed protein targets prediction and investigated interacting signal pathways. Furthermore, animal experiment was carried out to evaluate its efficacy and real-time quantitative polymerase chain reactions (RT-qPCR) analysis was explored the mechanism of action.
The detail information on alkaloids from S. flavescens were obtained from a handful of public databases on the basis of oral bioavailability (OB ≥ 30%) and drug-likeness (DL ≥ 0.18). Then, correlations between compounds and protein targets were linked using the STRING database, and targets associated with eczema were gathered by the GeneCards database. Human genes were identified and subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) functional enrichment analysis. Particularly, matrine, the crucial alkaloid from S. flavescens, was estimated using a 2,4-dinitrochlorobenzene (DNCB)-induced eczema Kunming (KM) mice model, administered (50 mg/kg and 10 mg/kg) to mice for 22 days. On the last day, the activities of serum tumor necrosis factor α (TNF-α), interleukin-4 (IL-4) and histopathologic examinations were determined. For further to elucidate the mechanisms, the mRNA levels of TNF-α, STAT3, TP53, AKT1, IL-6, JUN and EGFR in dorsal skin tissues were also tested.
Network analysis collected and identified 35 alkaloids from S. flavescens. Among them, in total 10 dominating alkaloids, including matrine, oxymatrine, sophoridine, sophocarpine, oxysophocarpine, allomatrine, sophoramine, anagyrine, cytisine and N-methylcytisine. And 71 related targets were provided of alkaloids for the treatment of eczema from S. flavescens. Furthermore, matrine dose-dependently (50 or 10 mg/kg, 22 days, apply to dorsal skin) remarkable decreased the serum levels of TNF-α and IL-4, and significantly alleviated the skin lesions. The effects of 50 mg/kg of matrine were almost identical to those of 200 mg/kg of the positive drug dexamethasone (DXM). The further RT-qPCR analyses could reveal that matrine down-regulate TNF-α, STAT3 and TP53 at transcriptional level in dorsal skin tissues.
Pharmacological network analysis can utilize to illuminate the pharmacodynamic substances and the potential molecular mechanism of S. flavescens for treating eczema. Matrine, as the crucial alkaloid from S. flavescens, could be a promising drug candidate for the treatment of eczema ailment.
苦参是一种常用于治疗皮肤疾病、腹泻、阴道瘙痒和炎症性疾病的传统中药。特别是苦参根与其他草药的组合主要用于临床应用中治疗湿疹。然而，苦参治疗湿疹的整体网络药理学机制尚未得到研究。
为了研究苦参生物碱治疗湿疹的网络药理学潜在作用，我们研究了生物碱，进行了蛋白质靶标预测，并研究了相互作用的信号通路。此外，还进行了动物实验来评估其疗效，并通过实时定量聚合酶链反应 (RT-qPCR) 分析探讨其作用机制。
根据口服生物利用度 (OB≥30%)和药物相似性 (DL≥0.18)，从少数公共数据库中获取苦参中的生物碱的详细信息。然后，使用 STRING 数据库将化合物和蛋白质靶标之间的相关性联系起来，并从 GeneCards 数据库中收集与湿疹相关的靶标。鉴定人类基因，并进行京都基因与基因组百科全书 (KEGG) 通路分析和基因本体论 (GO) 功能富集分析。特别是苦参中的关键生物碱苦参碱，通过 2,4-二硝基氯苯 (DNCB) 诱导的湿疹昆明 (KM) 小鼠模型进行评估，以 50mg/kg 和 10mg/kg 的剂量给小鼠给药 22 天。在最后一天，测定血清肿瘤坏死因子-α (TNF-α)、白细胞介素-4 (IL-4) 和组织病理学检查的活性。为了进一步阐明机制，还测试了背部皮肤组织中 TNF-α、STAT3、TP53、AKT1、IL-6、JUN 和 EGFR 的 mRNA 水平。
网络分析收集并鉴定了苦参中的 35 种生物碱。其中，苦参碱、氧化苦参碱、槐定碱、槐果碱、氧化槐果碱、金雀花碱、苦参胺、野靛碱、烟碱和 N-甲基烟碱等 10 种主要生物碱。苦参生物碱治疗湿疹的 71 种相关靶标。此外，苦参碱（50 或 10mg/kg，22 天，涂抹于背部皮肤）剂量依赖性地显著降低了血清 TNF-α和 IL-4 的水平，并显著缓解了皮肤损伤。50mg/kg 苦参碱的作用几乎与阳性药物地塞米松 (DXM) 200mg/kg 的作用相同。进一步的 RT-qPCR 分析表明，苦参碱在背部皮肤组织中转录水平下调 TNF-α、STAT3 和 TP53。
药理学网络分析可用于阐明苦参治疗湿疹的药效物质和潜在分子机制。苦参碱作为苦参中的关键生物碱，可能是治疗湿疹疾病的有前途的候选药物。
