基于血清代谢组学与网络药理学研究咳喘停穴位贴敷治疗哮喘的效应物质与作用机制

Anti-asthma components and mechanism of Kechuanting acupoint application therapy: based on serum metabolomics and network pharmacology

基于血清代谢组学和网络药理学研究咳喘停穴位贴敷治疗哮喘的效应物质及作用机制。将60例长期使用低剂量吸入性糖皮质激素-福莫特罗(ICS-formoterol)的哮喘患者随机分为西药组(30例)和西药+咳喘停组(30例),另纳入30名健康人为健康对照组。其中西药组予以低剂量ICS-formoterol治疗,西药+咳喘停组在西药基础上进行咳喘停穴位贴敷,健康对照组不进行任何干预。哮喘患者在治疗前、后进行哮喘控制测试(ACT)评分、肺功能指标第1秒用力呼气容积(FEV1)、呼气流量峰值(PEF)等疗效评价。采用气相色谱-质谱联用的代谢组学技术,结合多元数据统计筛选咳喘停穴位贴敷治疗哮喘的潜在生物标志物,进一步分析相关代谢通路。采用UPLC/LTQ-Orbitrap-MS技术结合网络药理学,构建咳喘停穴位贴敷治疗哮喘的“成分-靶点-通路”网络,探讨咳喘停穴位贴敷治疗哮喘的效应物质及作用机制。ACT评分、FEV1、PEF结果表明咳喘停穴位贴敷治疗哮喘具有较好的疗效。血清代谢组学共筛选出10个咳喘停穴位贴敷治疗哮喘的内源性生物标志物,获取了甘氨酸、丝氨酸和苏氨酸代谢,乙醛酸和二羧酸代谢等代谢通路。UPLC/LTQ-Orbitrap-MS鉴定了咳喘停穴位贴敷的51个化学成分。网络药理学分析表明咳喘停穴位贴敷治疗哮喘主要作用于SRC、MMP9等靶蛋白,与代谢途径、PI3K-Akt、MAPK、钙信号等通路密切相关;咳喘停穴位贴敷中芥子碱硫氰酸盐、延胡索甲素、氢化小檗碱等14个治疗哮喘的潜在效应物质。该研究通过血清代谢组学和网络药理学初步揭示了咳喘停穴位贴敷治疗哮喘的效应物质及作用机制,为深入研究作用机制和临床开发应用奠定理论基础。

This study aims to explore the anti-asthma components and mechanism of Kechuanting acupoint application therapy(KAAT) based on serum metabolomics and network pharmacology. A total of 60 asthma patients who had used low-dose inhaled corticosteroids-formoterol(ICS-formoterol) for a long time were randomized into the western medicine group(low-dose ICS-formoterol) and western medicine+Kechuanting group(KAAT+low-dose ICS-Formoterol), 30 in either group. In addition, 30 healthy people were included as the control(no intervention). The asthma control test(ACT) score, forced expiratory volume in 1 second(FEV1), and peak expiratory flow(PEF) were measured in the western medicine group and western medicine+Kechuanting group before and after treatment. The potential biomarkers of KAAT in the treatment of asthma were screened by gas chromatography-mass spectrometry combined with multivariate analysis, and the related metabolic pathways were further analyzed. UPLC/LTQ-Orbitrap-MS, together with network pharmacology, was employed to construct the component-target-pathway network. Thereby, the effective components and me-chanism of KAAT in the treatment of asthma were clarified. According to the ACT score, FEV1, and PEF, KAAT was effective in the treatment of asthma. A total of 10 endogenous biomarkers of KAAT in the treatment of asthma were screened by serum metabolomics, and the pathways of the metabolism of glycine, serine and threonine, and the metabolism of glyoxylic acid and dicarboxylic acid were obtained. UPLC/LTQ-Orbitrap-MS identified 51 chemical components of KAAT: 24 flavonoids, 11 alkaloids, 8 phenols, 2 diterpenoids, 2 triterpenoids, 2 glycosides, and 2 aldehydes. Network pharmacology analysis suggested that KAAT mainly acted on serum crea-tinine(SRC), matrix metalloproteinase 9(MMP-9), and other target proteins. The treatment was closely related to metabolic pathway, phosphatidylinositol 3-kinase-protein kinase B(PI3 K-Akt), mitogen-activated protein kinase(MAPK), and calcium signaling pathway. Sinapine thiocyanate, corydaline, dihydroberberine, stylopine, leonticine, N-methyl tetrahydroberberine, kaempferide, erio-dictyol, quercetin, catechin, 6-gingerol, 6-shogaol, ingenol, and luteolin may be potential effective compounds of KAAT in the treatment of asthma. This study preliminarily revealed that the effective components and mechanism of KAAT in treatment of asthma based on serum metabolomics and network pharmacology. It lays a theoretical foundation for in-depth study of the mechanism and clinical development and application.