基于UPLC-Q-Orbitrap HRMS、网络药理学与体外实验的潞党参入血抗氧化成分改善心肌缺血再灌注损伤机制探究OA

Objective:To investigate the mechanisms through which the blood-absorbed antioxidant components of Codonopsis Radix from Shanxi ameliorate myocardial ischemia-reperfusion injury(MIRI)using ultra-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap HRMS),network pharmacology,and cell experiments.Methods:UPLC-Q-Orbitrap HRMS was employed to compare the Codonopsis Radix extract and residual components obtained after riboflavin-methionine photolysis treatment,thus rapidly identifying the blood-absorbed antioxidant components of Codonopsis Radix.Network pharmacology and GEO database were employed to screen the key active components and differentially expressed genes.Molecular docking was then performed to validate their binding activities.H9c2 cardiomyocytes exposed to hypoxia-reoxygenation were used to validate the key pathways and targets of Codonopsis Radix in ameliorating MIRI.Results:Nineteen blood-absorbed antioxidant components were qualitatively identified in Codonopsis Radix.Through network pharmacology,seven core components were identified,along with six key targets.Among them,the binding energy of tangshenoside Ⅰ with each key target was less than-8.5 kcal·mol-1,indicating strong binding activity.Cell experiments revealed that the high-dose Codonopsis Radix extract significantly enhanced the superoxide dismutase(SOD)activity,reduced the levels of lactate dehydrogenase(LDH)and malondialdehyde(MDA),and increased cell survival rate.At the same time,it stimulated overexpression of hypoxia-inducible factor-1 alpha(HIF-1α)and downregulated the protein level of nuclear factor-kappa B1(NF-κB1),which inhibited NF-κB signaling pathway activation and ultimately suppressed the expression of the downstream inflammatory cytokine tumor necrosis factor-alpha(TNF-α).Tangshenoside Ⅰ,the core antioxidant component,also exerted a similar inhibitory effect on oxidative stress and normalized expression of HIF-1α,NF-κB1,and TNF-α,but with slightly weaker efficacy.Conclusion:The construction of the component-disease-pathway-target network and cell experiments identified the active components of Codonopsis Radix that can effectively inhibit oxidative stress and ameliorate ischemia-reperfusion injury in H9c2 cardiomyocytes.Furthermore,Codonopsis Radix does not rely on the potent effect of a single component but forms an overall regulatory network through the synergistic effects of multiple components and targets,thereby achieving a comprehensive intervention effect.