Study On Hepatotoxicity Mechanism Of Asarum Powder Based On Metabolomics And Transcriptomics

Objective: The volatile components of Asarum were determined using solid-phase microextraction technology,and the mechanism of hepatotoxicity caused by Asarum was predicted by network toxicology.Metabolomics and transcriptomics combined with biochemical methods were applied to study hepatotoxicity of Asarum powder and explore its hepatotoxicity mechanism.Methods: The volatile components of Asarum was determined by solid phase microextraction combined with GC-MS.The target of the measured volatile components was predicted by Swiss Target Prediction.The target of liver injury or liver toxicity was obtained by using Dis Ge NET.The mechanism of Asarum-induced liver toxicity was predicted by constructing network of compound-target,disease-target,compound-target-pathway.80 SD rats were randomly divided into four groups: control group,Asarum low-dose group(0.27 g/kg),Asarum medium-dose group(0.81 g/kg),and Asarum high-dose group(1.35 g/kg).There were 20 rats in each group.The rats were administered orally for 28 days,and the control group was given an equal volume of distilled water.The weight changes of rats in each group were recorded separately.After gavage,the serum was measured to determine liver function;the liver was weighed to calculate the liver coefficients;liver tissue was taken for HE staining to observe the morphological and histological changes;and then livers were tested for metabolomics.The differential metabolites were screened through multivariate statistical analysis method,and then the relevant metabolic pathways were found out.The livers of the high-dose group and the control group were sequenced to find differentially expressed genes;then GO enrichment analysis and pathway enrichment analysis were performed.Finally,Met Scape analysis was used to integrate the data of the screened differential metabolites and differentially expressed genes for exploring the mechanism of Asarum-induced liver toxicity.Results:(1)31 volatile components and 248 potential targets were detected in chemical composition analysis.There were 11 targets intersecting with liver toxicity.11 targets were mainly enriched in 5 metabolic pathways including PPAR signal pathway,chemical carcinogenesis,bile secretion,porphyrin and chlorophyll metabolism,metabolism of xenobiotics by cytochrome P450.(2)Compared with the control group,the weight of rats in each administration group decreased and the organ coefficients became larger,especially in the high-dose group(P<0.05).Liver function results showed that ALT,AST,TBil,and ALP increased,and TP decreased,and liver changes were greatest in the high-dose group.The HE staining results showed that Asarum in different dose groups showed different degrees of liver damage,manifested as balloon-like degeneration,loose cytoplasm,and punctate necrosis.(3)According to the GC-MS spectrum,14 differential metabolites were screened.Compared with the control group,lactic acid,alanine,aspartic acid,phosphoglycolic acid,taurine,xylose,sorbitol,and inositol were increased in livers of low,medium and high-dose rats;levels of proline,methionine,ribitol,spermidine,trehalose,and maltotriose were decreased.Pathway enrichment results show that the main metabolic pathways affected by Asarum were aminoacyl-t RNA biosynthesis,phosphoinositide metabolism,galactose metabolism,alanine,aspartic acid and glutamic acid metabolism,arginine and proline metabolism,metabolism of taurine and hypotaurine.(4)Transcriptome sequencing results revealed that 434 genes were significantly changed,of which 214 genes were significantly up-regulated and 220 genes were significantly down-regulated.These differentially expressed genes are mainly related to circadian rhythm,p53 signaling pathway,metabolic pathway,steroid biosynthesis,bile secretion and other pathways.Conclusion:(1)Long-term use of high-dose Asarum powder has damage to the liver function and tissue morphology of rats,and this damage is positively related to the dose,that is,the higher the dose,the greater the degree of liver damage.(2)The mechanism of toxicity of Asarum on the liver is closely related to bile acid metabolism,amino acid metabolism,inositol phosphate metabolism,galactose metabolism,circadian rhythm,and p53 signaling pathway.