Toxicity attenuation processing technology and mechanism of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction / 中国中药杂志

This study explored toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction for the first time, and further explored its detoxification mechanism. Nine processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction were prepared by orthogonal experiment with three factors and three levels. Based on the decrease in the content of the main hepatotoxic component diosbulbin B before and after processing of Rhizoma Dioscoreae Bulbiferae by high-performance liquid chromatography, the toxicity attenuation technology was preliminarily screened out. On this basis, the raw and representative processed products of Rhizoma Dioscoreae Bulbiferae were given to mice by gavage with 2 g·kg~(-1)(equival to clinical equivalent dose) for 21 d. The serum and liver tissues were collected after the last administration for 24 h. The serum biochemical indexes reflecting liver function and liver histopathology were combined to further screen out and verify the proces-sing technology. Then, the lipid peroxidation and antioxidant indexes of liver tissue were detected by kit method, and the expressions of NADPH quinone oxidoreductase 1(NQO1) and glutamate-cysteine ligase(GCLM) in mice liver were detected by Western blot to further explore detoxification mechanism. The results showed that the processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reduced the content of diosbulbin B and improved the liver injury induced by Rhizoma Dioscoreae Bul-biferae to varying degrees, and the processing technology of A_2B_2C_3 reduced the excessive levels of alanine transaminase(ALT) and aspartate transaminase(AST) induced by raw Rhizoma Dioscoreae Bulbiferae by 50.2% and 42.4%, respectively(P<0.01, P<0.01). The processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reversed the decrease protein expression levels of NQO1 and GCLM in the liver of mice induced by raw Rhizoma Dioscoreae Bulbiferae to varying degrees(P<0.05 or P<0.01), and it also reversed the increasing level of malondialdehyde(MDA) and the decreasing levels of glutathione(GSH), glutathione peroxidase(GPX), and glutathione S-transferase(GST) in the liver of mice(P<0.05 or P<0.01). In summary, this study shows that the optimal toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction is A_2B_2C_3, that is, 10% of Paeoniae Radix Alba decoction is used for moistening Rhizoma Dioscoreae Bulbiferae and processed at 130 ℃ for 11 min. The detoxification mechanism involves enhancing the expression levels of NQO1 and GCLM antio-xidant proteins and related antioxidant enzymes in the liver.

The metabolic profile of serum fatty acids in rats with Rhizoma Dioscoreae Bulbiferae-induced liver injury / 药学学报

Rhizoma Dioscoreae Bulbiferae is a traditional Chinese medicine with hepatotoxicity, but the metabolic profile of fatty acids has not been identified in the rats with liver injury. In this project, a gas chromatography-mass spectrometry method was applied to simultaneous quantification of 16 non-esterified fatty acids (NEFA) and esterified fatty acids (EFA) in the serum of control, ethanol extraction of Rhizoma Dioscoreae Bulbiferae (ethanol extraction, ET) and diosbulbin B (DB)-treated rats. Meanwhile, the change of fatty acid metabolic profile of liver injured rats was analyzed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The results of NEFA concentration indicated that the serum concen-trations of palmitic acid (C16:0), stearic acid (C18:0), palmitoleic acid (C16:1n7), oleic acid (C18:1n9), vaccenic acid (C18:1n7), linoleic acid (C18:2n6), linolenic acid (C18:3n3), eicosatrienoic acid (C20:3n6), arachidonic acid (C20:4n6) and docosahexaenoic acid (C22:6n3) in DB-treated rats decreased significantly, while that of C18:2n6 and C20:3n6 obviously increased and that of C20:4n6 and C22:6n3 noticeably dropped in ET-treated rats when compared with control. Furthermore, the results of EFA concentration illus-trated that the serum concentrations of C16:0, C18:0, C20:4n6, C22:6n3 and eicosapentaenoic acid (C20:5n3) in two toxic groups were remarkably decreased when compared with control. The fatty acid meta-bolic profiles of the two toxic groups exhibited significant difference from the normal levels, and the degree of deviation of ET group was higher than that of DB group. More importantly, the results of PLS-DA showed that C20:4n6 and C22:6n3 were important indicators of the hepatotoxicity induced by ET and DB, and the serum concentrations of the two fatty acids had good correlation with the levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin using Pearson's correlation analysis and canonical correlation analysis (CCA). Therefore, C20:4n6 and C22:6n3 were identified as potential biomarkers of ET and DB-induced liver injury. The project can provide a foundation for furture investigation of molecular mechanism of hepato-toxicity caused by Rhizoma Dioscoreae Bulbiferae.

Protection of Herba Scutellariae Barbatae against hepatotoxicity induced by Rhizoma Dioscoreae Bulbiferae and its mechanism / 药学学报

This study was conducted to test the protective activity of ethanol extract of Herba Scutellariae Barbatae (SE) against hepatotoxicity induced by Rhizoma Dioscoreae Bulbiferae in mice and its mechanism. SE was orally given to mice at various doses, and ethyl acetate fraction of Rhizoma Dioscoreae Bulbiferae (EF, 450 mg·kg-1) was also orally given at the same time. After 11 days, serum levels of alanine/aspartate aminotransferase (ALT/AST), alkaline phosphatase (ALP), total protein (TP) and albumin (ALB) were measured, and liver histological examination was conducted. Liver glutathione (GSH) amount, myeloperoxidase (MPO) activity and serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interferon-γ (IFN-γ) levels were measured. The expression of heme oxygenase-1 (HO-1), inhibitor of kappa B (IκB) and nuclear factor κB (NF-κB) p65 were determined by Western blot. The results showed that SE (200 mg·kg-1) reversed EF-induced changes of serum ALT, AST, ALP, TP and ALB. Liver histology also suggests the protection of SE against EF-induced liver injury. SE reduced the increased MPO activity in liver and TNF-α, IL-6, IFN-γ contents in serum, and blocked the decrease in IκB expression and subsequent increase in phosphorylation and nuclear translocation of p65 induced by EF. EF increased liver GSH amount and heme oxygenase-1 (HO-1) protein expression in mice. SE increased liver GSH amount, but decreased the expression of HO-1. All those results suggest that SE alleviates liver injury induced by consecutive administration of EF by alleviating inflammatory injury via inhibiting NF-κB signaling pathway and elevating antioxidant capacity.

Metabonomic study on early biomarkers of hepatic injury induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae in rats based on 1H-NMR / 中国药理学与毒理学杂志

OBJECTIVE Dynamics of serum and urine metabolites in hepatic injury rats induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae(RDB)was investigated by 1H-NMR-based metabo?nomic methods in order to discover early biomarkers of liver toxicity induced by RDB. METHODS Rats were ig adminisetred with RDB at a dose of 5 g·kg-1 for 28 d. Rats were sacrificed 3,7,14 and 28 d af?ter RDB administration,as well as after a recovery period,respectively. Blood was taken for routine bio?chemical analysis by an automatic biochemical analyzer. Liver/body mass indexes were calculated ,and liver pathological changes were observed with hematoxylin-eosin staining. Urine samples were collected before and 3,7,14 and 28 d after RDB administration,respectively,as well as after withdrawal. Metabo?nomic analysis was carried out for serum and urine samples. Principal component analysis and orthogonal partial least squares-discriminant analysis were used for screening and identifiying early biomarkers. RESULTS Compared with the control group,total bilirubin (TB) and total cholesterol (TC) values were increased in 3-28 d in RDB group(P<0.05,P<0.01). Total bile acid(TBA)was elevated in 7-28 d (P<0.05,P<0.01). TB,TC and TBA became normal after discontinuation with RDB. There was no significant difference between RBD-treated group and control group in the activity of glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase,and the content of glucose also was not different between the two groups. The ratio of liver/body mass was elevated at 3-28 d(P<0.01)but returned to normal after withdraval of RDB. The enlargement and necrosis of hepatocytes were observed 7 d after RDB administration,and lesion degree was aggravated with the extension of RDB delivery time. Meta?bonomic analysis showed that the serum lipids (low density lipoprotein/very low density lipoprotein (LDL/VLDL),glutamic acid,choline phosphate and glycerolphosphatecholine were increased in the early stage. Pyruvate and N-acetylglutamate were decreased in urine. These metabolites became normal 7 d after discontinuation with RDB. CONCLUSION The serum lipids (LDL/VLDL),glutamic acid,glycerol phosphate choline,as well as urine pyruvic acid salt and N-acetyl glutamate may be used as the early biomarkers for liver toxicity induced by RDB.

Sensitive genes screening of mice liver injury induced by hepatotoxicant and their biological function analysis / 中华中医药杂志

Objective:To scan the common differentially expressed genes in mice liver tissue following subacute exposure to hepatotoxicants,including alcohol,carbon tetrachloride,BCG & LPS,the alcohol extracts of Rhizoma Dioscoreae Bulbiferae and Senecio scandens Buch.-Ham.,and apply to the rapid toxic evaluation of TCM and other xenobiotics.Methods:The gene expression profiles of mice liver tissue,after respective administration of alcohol,CCl4,BCG associated with LPS,the alcohol extracts of Rhizoma Dioscoreae Bulbiferae and Senecio scandens Buch.-Ham.to healthy mice,were analyzed by use of mouse genome cDNA microarray.The correlations between the common differential expressed genes and the liver injury were analyzed based on the biological functions of those differentially expressed genes.Results:Among all of the five drug administration groups,there were 7 known function genes(BC05200,NM-019410,NM-173019,AB028272,AK088925,AK030862 and AK088816)and 1 unknown function gene(BC069871)differentially expressed.Beside AK088816,the other 7 genes were all down-regulate expressed.Conclusion:The common differentially expressed genes participate in the process of saccharometabolism,apoptosis,cell growth cycle,cytoskeleton and signal transmission,protein folding and protein ubiquitin.The abnormal expression of the common genes were closely correlated to the liver,and might be important for rapid hepatoxicity elucidation of traditional Chinese medicine and other xenobiotics.