液质联用结合网络药理学探究少腹逐瘀汤抗宫颈癌活性机理

The Anti-Cervical Cancer Mechanism of Shaofu Zhuyu Decoction Based on Liquid Chromatography Mass Spectrometry Combined with Network Pharmacology

采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-Exactive Orbitrap MS)技术,结合网络药理学和分子对接,对少腹逐瘀汤发挥抗宫颈癌作用的重要成分、靶点和通路进行预测分析,并通过细胞实验初步探究其作用机制。经鉴定发现苯丙素类、黄酮类、萜类、生物碱类、有机酸类、挥发油类、苯酞类、香豆素类、苯醛类、氨基酸类、酚类和其他类等共89种化合物,网络药理学和分子对接深入挖掘得到少腹逐瘀汤抗宫颈癌的7个重要成分、8个核心靶点和10条宫颈癌相关信号通路。细胞实验表明,给药组HeLa细胞与空白对照组相比,生存率逐渐降低,细胞凋亡率逐步提高,呈浓度依赖性(P<0.05)。少腹逐瘀汤可以提高Bax、Cyto c、Caspase 9和Caspase 3表达水平,降低PI3K、AKT和Bcl-2表达水平。少腹逐瘀汤可能通过槲皮素、芍药苷、黄芩素、山奈酚、沙立朴吩、丁香酚和鞣花酸等成分作用于TP53、AKT1和EGFR等靶点发挥抗宫颈癌作用,其机制与PI3K/AKT和Bcl-2家族蛋白信号通路的调控密切相关,充分体现了多成分、多靶点和多通路的特点。

The objective of this study was to elucidate the chemical components of Shaofu Zhuyu Decoction(SFZYD)and to investigate its potential anti-cervical cancer mechanism.This study applied UHPLC-QExactive Orbitrap MS technique,in combination with network pharmacology and molecular docking,to predict and analyze the important components,targets,and pathways of SFZYD involved in its anti-cervical cancer effect.CCK-8 assay,Hoechst 33342 staining assay,flow cytometry,and WB assay were employed to detect the apoptosis and related gene protein expression levels of HeLa cells.The results revealed that 89 compounds,belonging to phenylpropanoids,flavonoids,terpenoids,alkaloids,organic acids,volatile oils,phthalides,coumarins,benzaldehydes,amino acids,phenols,and others,were identified.Through in-depth exploration of network pharmacology and molecular docking,seven key components,eight core targets,and ten cervical cancer related signaling pathways of SFZYD were screened out to combat cervical cancer.Cell experiments demonstrated that compared with the blank control group,the survival rate of HeLa cells in the treatment group gradually decreased,and the apoptosis rate gradually increased in a concentration-dependent manner(P<0.05).Furthermore,SFZYD could increase the expression levels of Bax,Cyto c,Caspase 9,Caspase 3,and decrease the expression levels of PI3K,AKT,and Bcl-2.This suggested that SFZYD might exert its anti-cervical cancer effect on targets such as TP53,AKT1,and EGFR through components such as quercetin,paeoniflorin,baicalein,kaempferol,ST057701,eugenol,and ellagic acid.Its mechanism was closely associated with the regulation of PI3K/AKT pathway and Bcl-2 family,fully reflecting the characteristics of multi-component,multi-target,and multi-pathway.