Research Of Effective Components In Huangqi Decoction Against Liver Fibrosis

Objective:Huangqi Decoction（HQD）is a classic prescription for the treatment of liver fibrosis,but its effective ingredients are still unclear.The purpose of this study is to explore the effective components of HQD against liver fibrosis based on the strategies of analyzing of the chemical composition combined with efficacy verification in vivo and in vitro.And the results could help to analyze the effective material basis of HQD and provide new methods for investigation of traditional Chinese medicine（TCM）formulas.Methods:（1）Using UPLC-LTQ-Orbitrap method,the main components of HQD were characterized.（2）The same method was used to qualitatively analyze the absorbed components and metabolites in plasma and urine samples of healthy volunteers after oral administration of HQD.（3）The effects of the absorption components of HQD on cell apoptosis were evaluated in cultured LX-2 cells;and the expression of alpha-smooth muscle actin（a-SMA）and type I collagen（Col I）in TGF-β1 induced cell fibrosis model were investigated.（4）The mouse liver fibrosis model inducedby intraperitoneal injection of 10%CCl₄was used to investigate the effect of combination of multiple effective components（MCHD）.The serum level of alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,and liver morphology and hydroxyproline content that influenced by MCHD were further investigated.Results:（1）Qualitative analysis of HQD extract identified 64 representative components,mainly including triterpene saponins and flavonoids.Among them,34ingredients are derived from Astragali Radix and 31 are derived from Radix et Rhizoma Glycyrrhizae.（2）A total of 24 absorption components and 17 metabolites were further identified in plasma and urine samples of healthy volunteers after oral administration of HQD.Among them,24 prototypes and 17 metabolites were identified in human plasma;9 prototypes and 15 metabolites were identified in human urine.（3）In vitro experiments revealed that,treatment with astragaloside IV,cycloastragenol,glycyrrhizic acid,glycyrrhetinic acid,liquiritigenin,and isoliquiritigenin markedly decreased the expression ofα-SMA;and treatment with cycloastragenol,calycosin-7-glucoside,formononetin,glycyrrhetinic acid,liquiritin orisoliquiritin significantly decreased the m RNA expression of Col I;while treatment with calycosin,liquiritigenin,isoliquiritigenin,cycloastragenol and glycyrrhetinic acid significantly accelerated the apoptosis of LX-2 cells.（4）Compared with model group,treatment with MCHD significantly reduced the serum levels of ALT and AST of fibrotic mice.Additionally,MCHD also ameliorated the liver collagen fibril deposition and hydroxyproline content in CCl₄-induced liver fibrosis mice,and improved liver injury and fibrosis.Conclusion:This study analyzed and identified the main components of HQD in vivo and in vitro.The combination of the multiple effective components of HQD has a profound significant anti-fibrosis effect.The current research provides new methods and strategy for exploring effective ingredients of traditional Chinese herbal formula.