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Mechanism of Sangqi Qingxuan Liquid in Alleviating Vascular Endothelial Injury in Hypertension Focuses on β-Catenin

摘要Objective:To explore the main components and potential mechanisms of Sangqi Qingxuan Liquid in the treatment of arterial vascular endothelial cells(AVECs)injury in hypertension through network pharmacology.Methods:Traditional Chinese Medicine Systems Pharmacology and Analysis Platform(TCMSP)and Traditional Chinese Medicine Integrated Database(TCMID)were used to screen the active components of Sangqi Qingxuan Liquid(SQQX),which met the oral utilization rate and drug similarity criteria.An active component-target network was constructed using Cytoscape 3.6 software.A protein-protein interaction(PPI)network of targets associated with SQQX treatment for hypertension was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database.The Metascape database was used to perform enrichment analysis of gene ontology biological functions and MSigDB pathway enrichment analysis of proteins in the PPI network.Further analysis of the main components of SQQX was performed using UPLC-MS.Based on the results of network pharmacology,the mechanism of SQQX to improve the injury of AVECs in hypertension was verified through lentiviral transfection by Wnt/β-catenin signaling pathway.AVECs induced by angiotensin Ⅱ(Ang Ⅱ)was used to establish a model of endothelial function injury in hypertension.Cell viability,intracellular nitric oxide content,malonaldehyde content,and superoxide dismutase activity were measured to determine the optimal induction conditions.The optimal intervention conditions for SQQX were determined based on cell viability,cellular DNA activity,and the gradient method.The cells were further divided into blank,model,overexpression lentivirus negative control,overexpression lentivirus,overexpression lentivirus+SQQX intervention(2.47 mg/mL,12 h),inhibition lentivirus negative control,inhibition lentivirus,and inhibition lentivirus+SQQX intervention(2.47 mg/mL,12 h)groups.Finally,quantitative real-time PCR and Westem blotting were performed to analyze the molecular mechanisms of SQQX in the Wnt/β-catenin signaling pathway.Results:The main SQQX components were betaine,buddleoside,and chlorogenic acid,in descending order.Network pharmacology analysis screened 12 pathways associated with the hypertensive vascular endothelium.The results showed that 1 μ mol/L for 12 h was the optimal condition for Ang Ⅱ to induce AVECs injury,and 2.47 mg/mL SQQX intervention for 12 h was the optimal condition for treating AVECs injury.In the experimental validation based on the interaction network of the Wnt/β-catenin signaling pathway,SQQX significantly decreased the expressions of β-catenin,Smad2,peroxisome proliferator-activated receptors(PPARs),endothelial nitric oxide synthase(eNOS),and endothelin-1(ET-1)caused by the β-catenin overexpression lentivirus(P<0.05 or P<0.01).The function of vascular endothelial cells can be improved by the β-catenin inhibition lentivirus,and no obvious changes were observed after further intervention with SQQX.Conclusion:SQQX may protect against AVECs injury by regulating the Wnt/β-catenin signaling pathway.

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