背景:脓毒症是一种危及生命的器官功能障碍,严重威胁人类健康。临床和实验结果均证实,中医,如黄芩,具有抗炎作用。这为脓毒症的医治供给了新的思绪。本研究基于网络药理学,系统分析黄芩治疗脓毒症的作用机制,RNA测序和分子对接。
方法:使用BulkRNA测序对脓毒症患者和健康志愿者进行基因表达分析。对结果进行质量控制后,分析差异表达基因(DEGs)。利用中药系统药理学数据库和分析平台(TCMSP)对黄芩的有效成分和作用靶点进行鉴定。对疾病-药物交叉靶标进行基因本体论(GO)和蛋白质-蛋白质相互作用(PPI)分析。在GEO数据库的帮助下,对交叉指标进行生存分析和Meta分析,评价预后价值,筛选核心指标。随后,单细胞RNA测序用于确定核心靶标在细胞内的位置。最后,在这项研究中,通过分子对接实验进一步阐明黄芩有效成分与相应靶标的相互关系。
结果:黄芩的有效成分有72种,和50个常见的药物和疾病的目标。GO和PPI分析表明,交叉目标主要参与对化学胁迫的响应,对氧气水平的反应,对药物的反应,调节免疫系统的过程。生存分析显示PRKCD,EGLN1和CFLAR与脓毒症预后呈正相关。Meta分析发现这3个基因在脓毒症幸存者中高表达,而在非幸存者中卑微。PRKCD主要在巨噬细胞中发现,而EGLN1和CFLAR在免疫细胞中广泛表达。活性成分芹菜素调节CFLAR表达,黄芩素调节EGLN1表达,和Wogonin调节PRKCD表达。分子对接研究表明,黄芪的三种活性成分与其相应的靶标具有良好的结合活性。
结论:芹菜素,黄芩素和Wogonin,黄芩的重要活性成分,通过调节其靶标CFLAR的表达来产生抗脓毒症作用,EGLN1和PRKCD。
BACKGROUND: Sepsis is a life-threatening organ dysfunction, which seriously threatens human health. The clinical and experimental results have confirmed that Traditional Chinese medicine (TCM), such as Scutellariae Radix, has anti-inflammatory effects. This provides a new idea for the treatment of sepsis. This study systematically analyzed the mechanism of Scutellariae Radix treatment in sepsis based on network pharmacology, RNA sequencing and molecular docking.
METHODS: Gene expression analysis was performed using Bulk RNA sequencing on sepsis patients and healthy volunteers. After quality control of the results, the differentially expressed genes (DEGs) were analyzed. The active ingredients and targets of Scutellariae Radix were identified using The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Gene Ontology (GO) and Protein-Protein Interaction (PPI) analysis were performed for disease-drug intersection targets. With the help of GEO database, Survival analysis and Meta-analysis was performed on the cross-targets to evaluate the prognostic value and screen the core targets. Subsequently, single-cell RNA sequencing was used to determine where the core targets are located within the cell. Finally, in this study, molecular docking experiments were performed to further clarify the interrelationship between the active components of Scutellariae Radix and the corresponding targets.
RESULTS: There were 72 active ingredients of Scutellariae Radix, and 50 common targets of drug and disease. GO and PPI analysis showed that the intersection targets were mainly involved in response to chemical stress, response to oxygen levels, response to drug, regulation of immune system process. Survival analysis showed that PRKCD, EGLN1 and CFLAR were positively correlated with sepsis prognosis. Meta-analysis found that the three genes were highly expressed in sepsis survivor, while lowly in non-survivor. PRKCD was mostly found in Macrophages, while EGLN1 and CFLAR were widely expressed in immune cells. The active ingredient
Apigenin regulates CFLAR expression, Baicalein regulates EGLN1 expression, and Wogonin regulates PRKCD expression. Molecular docking studies confrmed that the three active components of astragalus have good binding activities with their corresponding targets.
CONCLUSIONS: Apigenin, Baicalein and Wogonin, important active components of Scutellaria Radix, produce anti-sepsis effects by regulating the expression of their targets CFLAR, EGLN1 and PRKCD.