Study On The Effective Substances And Mechanism Of Sanhuang Xiexin Decoction Against Hypercholesterolemia

Purpose: Hypercholesterolemia is a common metabolic disease,which is the main cause of cardiovascular and cerebrovascular diseases such as atherosclerosis,coronary heart disease and stroke.As a traditional Chinese compound preparation,Sanhuang Xiexin Decoction has the effects of clearing heat and detoxifying,purging fire and relaxing bowels,drying dampness and relieving heat.Modern pharmacological studies have shown that Sanhuang Xiexin Decoction has anti-inflammatory and lipid-lowering effects.The purpose of this study was to explore the effective substances and mechanism of Sanhuang Xiexin Decoction in the treatment of hypercholesterolemia,and to provide a new experimental basis for the clinical application of Sanhuang Xiexin Decoction in the treatment of hypercholesterolemia.Methods: Firstly,the cell and animal models were constructed to clarify the therapeutic effect of Sanhuang Xiexin Decoction on hypercholesterolemia.Then,UHPLC-QOrbitrap/MS combined with mz Cloud,Masslist,Chemspider and self-built database were used to comprehensively characterize the material groups in Sanhuang Xiexin Decoction.Based on this,the component-disease-target network was constructed.Through proteinprotein interaction network analysis and target-pathway network analysis,the key active components,important targets and action pathways of Sanhuang Xiexin Decoction were inferred.Based on UHPLC-Q-Orbitrap/MS,metabolomics studies were performed to determine the relevant metabolic pathways.Based on the comprehensive analysis of network pharmacology and metabolomics results,the common pathway was used as the main intervention pathway for Sanhuang Xiexin Decoction in the treatment of hypercholesterolemia.The key active components and corresponding targets were verified by molecular docking,and the pharmacokinetics of active components was studied.Results: Sanhuang Xiexin Decoction can significantly reduce the levels of TC,LDL-c and TG in Hep G2 cells with lipid accumulation,improve the level of HDL-c,and improve the levels of TC,LDL-c and TG in mice with hypercholesterolemia.Based on UHPLC-QOrbitrap/MS analysis,184 components were identified from Sanhuang Xiexin Decoction,including flavonoids,alkaloids,anthraquinones,phenols,glycosides,phenylpropanoids,etc.Nineteen prototype components were detected in plasma,including 9 flavonoids,4anthraquinones,5 alkaloids,1 organic acid,and 8 metabolites.Network pharmacology analysis showed that the main active components were emodin,aloe-emodin,wogonin,berberine,jatrorrhizine,etc.,involving 20 key targets such as JUN,SRC,AKT1,TNF,and tumor necrosis factor,P13K-Akt,sphingolipid metabolism pathway,interleukin-17 and other related signaling pathways.Metabolomics studies have shown that Sanhuang Xiexin Decoction may affect sphingolipid metabolism by regulating sphingosine and ceramide levels.It affects arginine and proline metabolism and arginine biosynthesis by regulating arginine and trans-4-hydroxy-D-proline levels.It affects tryptophan metabolism by regulating 5-hydroxyindole-3-acetic acid and kynurenine levels,and affects ether lipid metabolism and glycerophospholipid metabolism by regulating choline levels to improve hypercholesterolemia.Integrating the results of network pharmacology and metabolomics analysis,it is suggested that the active ingredient wogonin in Sanhuang Xiexin Decoction acts on AKT1,aloe-emodin,emodin and wogonin act on TNF and TP53,jatrorrhizine and berberine may act on e NOS to affect sphingolipid metabolism pathway,thus reducing cholesterol level.Molecular docking results showed that the active components had good affinity with key targets.The half-life of emodin,aloe-emodin,wogonin,jatrorrhizine and berberine in hypercholesterolemia rats was significantly delayed,and the peak time was also different.The bioavailability of aloe-emodin,wogonin,berberine and jatrorrhizine in hypercholesterolemia rats was higher than that in normal rats,which provided further reference for us to explore the effective substances of Sanhuang Xiexin Decoction in the treatment of hypercholesterolemia.Conclusion: In this paper,based on UHPLC-Q-Orbitrap/MS,the chemical constituents of Sanhuang Xiexin Decoction were comprehensively characterized in vitro and in vivo.Based on this,network pharmacology analysis was carried out.Combined with metabolomics research,it was found that the active ingredient wogonin in Sanhuang Xiexin Decoction may act on AKT1,aloe-emodin,emodin and wogonin may act on TNF and TP53,jatrorrhizine and berberine may act on e NOS to affect sphingolipid metabolism and achieve the effect of treating hypercholesterolemia.This paper systematically expounds the effective substances and mechanism of Sanhuang Xiexin Decoction in the treatment of hypercholesterolemia,and provides reference for the clinical application of Sanhuang Xiexin Decoction.