RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Liu-Shen-Wan (LSW) is one of the popular over-the-counter drugs in Asia, which contains realgar (As4S4), used for the treatment of upper respiratory tract inflammation and skin infections. However, the safety and potential risk of this arsenic remain unknown. AIM OF THE STUDY: The aim of this study was to determine total arsenic in tissue and investigate effects of regular dose and overdose LSW exposure on rat liver. MATERIALS AND METHODS: We used a target lipidomics approach to quantify inflammatory eicosanoids and employed ICP-MS to determine total arsenic in tissue. RESULTS: The results showed that oral administration of 8 and 40â¯mg/kg LSW (1 and 5 fold human-equivalent dose) induced light changes of liver lipidomic profile in rats, which was associated with anti-inflammatory function of LSW. In our recent report, we observed that 41 and 134â¯mg/kg realgar (40 and 132 fold human-equivalent dose) stimulated rat liver inflammation through up-regulation of pro-inflammatory LOX-derived, CYP-derived HETEs and COX-derived PGs. However, we found that LSW in the form of drug combination, containing 41 and 134â¯mg/kg realger, could not stimulate these similar inflammatory responses in rats, although the liver total arsenic levels of the realger and LSW groups were same. CONCLUSION: The downregulation of pro-inflammatory response showed that the LSW containing realger is safer than realger alone administrated to rats. These results suggested that Chinese medicines combination could reduce realgar-derived arsenic toxicity in rats.
Asunto(s)
Mezclas Complejas/efectos adversos , Inflamación/inducido químicamente , Inflamación/metabolismo , Medicina Tradicional China/efectos adversos , Animales , Antiinflamatorios/farmacología , Arsénico/efectos adversos , Mezclas Complejas/farmacología , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Lipidómica/métodos , Hígado/efectos de los fármacos , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
In order to evaluate the quality of Bufonis Venenum commercial herbs, a three-step qualitative and quantitative research study was performed. Firstly, we tried to identify small molecules and peptides in Bufonis Venenum using pre-fractionation chromatography and high-resolution mass spectrometry. The database search of the small molecules and peptides of Bufonis Venenum revealed that the dried venom consisted of free/conjugated-type bufadienolides and peptides with a mass range of 0.4-2 kDa. Secondly, we used partial least squares (PLS) multivariate statistical analysis to screen bufadienolides markers (VIP > 1.5) responsible for the anti-tumor cell activity of Bufonis Venenum, including 21 identified bufadienolides and 7 unknown compounds. It is noticeable that these bufadienolide markers could not be recognized by traditional HPLC-UV based spectrum-effect relationship analysis (correlation coefficient ranging from -0.24 to 0.40). Finally, we proposed a weight coefficient-based corrected total contents of 9 bufadienolides as a quality evaluation indicator, which had good correlation with inhibitory effects on tumor cells of commercial Bufonis Venenum. The correlation coefficient increased from 0.4 to 0.6. Thus, our pre-fractionation chromatography and mass spectrometry strategy had significant advancement over the traditional spectrum-effect relationship method for chemical marker identification. These results could be crucial and helpful in the development of a quality evaluation method that could reflect the pharmacological activity of Bufonis Venenum.
Asunto(s)
Antineoplásicos/farmacología , Bufanólidos/análisis , Bufanólidos/farmacología , Espectrometría de Masas , Péptidos/análisis , Péptidos/farmacología , Secuencia de Aminoácidos , Calibración , Línea Celular Tumoral , Humanos , Análisis de los Mínimos Cuadrados , Límite de Detección , Análisis Multivariante , Péptidos/química , Reproducibilidad de los Resultados , Bibliotecas de Moléculas Pequeñas/análisisRESUMEN
Arsenic sulfide compounds provide nearly all of the world's supply of arsenic. However, the risk of arsenic trisulfide exposure is still not fully investigated. Here, we systemically assessed the toxicology of As4S4 in rats by combining arsenic metabolite detection, routine testing and lipidomic profiling. It was revealed that the oral administration of As4S4 for two months increased the total arsenic content in the liver reaching a saturation level. Further analysis by anion exchange chromatography coupled with inductively coupled plasma mass spectrometry (ICP-MS) technology showed no trace of inorganic arsenic, but there was significant presence of dimethylarsinic acid (DMA), in the livers of rats. This arsenic metabolite was less toxic to rats and did not induce overt liver pathology and functional injury. In contrast, lipidomic profiling provided a comprehensive map of lipids and uncovered a more complex inflammatory response, exhibiting more sensitive change to arsenic exposure. We observed that metabolites of cyclooxygenase, including PGF2α, dhk PGF2α, 15k PGF2α, 8-iso-PGF2a, PGE2, dhk PGE2, PGD2, 15d-PGD2, and PGJ2, were significantly elevated. But mediators from lipoxygenase, cytochrome P450, docosahexaenoic acid, and eicosapentaenoic acid pathways were not markedly affected. In summary, we identified DMA as the predominant arsenic species in the livers of rats, and found cyclooxygenase-derived lipids as the inflammatory mediators before the development of overt liver injury for subchronic As4S4 exposure. These mediators could translate into potential metabolic biomarkers in early arsenic risk assessment and as targets for therapeutic intervention.
Asunto(s)
Arsenicales , Mediadores de Inflamación , Lipidómica , Lípidos/análisis , Sulfuros , Animales , Arsenicales/análisis , Biomarcadores/análisis , Biomarcadores/metabolismo , Eicosanoides/metabolismo , Mediadores de Inflamación/análisis , Mediadores de Inflamación/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Sulfuros/análisis , Sulfuros/toxicidadRESUMEN
A highly sensitive and rapid ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been developed and validated for simultaneous quantification of seven components in rat plasma and five components in rat tissues after oral administration of the extracts of different combination Radix Angelicae Sinensis-Radix Paeoniae Alba herb couple and has been applied to compare the different pharmacokinetics and tissue distribution properties of these bioactive components. The extracts of Radix Angelicae Sinensis (RAS), Radix Paeoniae Alba (RPA) and Radix Angelicae Sinensis-Radix Paeoniae Alba herb couple (RRHC) were orally administrated to rats, respectively. The concentrations of ferulic acid, caffeic acid, vanillic acid, ligustilide, paeoniflorin, albiflorin and oxypaeoniflorin in rat plasma and the concentrations of ferulic acid, vanillic acid, paeoniflorin, albiflorin and oxypaeoniflorin in tissues were determined by UPLC-MS/MS. The plasma samples were pretreated by protein precipitation with methanol and the tissue samples were homogenated with water and pretreated by protein precipitation with methanol. Chromatographic separation was performed on a C18 column using 0.1% formic acid-acetonitrile as mobile phase for gradient elution. A triple quadrupole (TQ) tandem mass spectrometry equipped with an electrospray ionization source was used as detector operating both in positive and negative ionization mode and operated by multiple-reaction monitoring (MRM) scanning. Noncompartmental pharmacokinetic parameters were calculated by DAS 2.0 program. The differences between each group were compared by SPSS 16.0 with Independent-Samples T-test. The pharmacokinetic parameters (such as Cmax, Tmax, T1/2, AUC0-T, MRT0-T, Vz/F or CLz/F) of all the detected components between the single herb (RAS or RPA) and herb pair (RRHP) showed significant differences (P<0.05). It indicated that the compatibility of RAS and RPA could alter the pharmacokinetics features of each component. Tissue distribution results showed that ferulic acid, vanillic acid, paeoniflorin, albiflorin and oxypaeoniflorin mostly distributed in liver and kidney both in herb couple and single herb distributed most in liver and kidney. Compared with single herb, RRHC could increase or decrease the concentrations of five components at different time points compared with the sing herb. The results indicated the method was successfully applied to the comparative study on pharmacokinetics and tissue distribution of different combination of RRHC in rats. The compatibility of two Chinese herbs could alter the pharmacokinetics and tissue distribution properties of major bio-active components in the single herb. The results might be helpful for further investigation of compatibility mechanism of RRHC.
Asunto(s)
Medicamentos Herbarios Chinos/farmacocinética , Paeonia/química , Extractos Vegetales/farmacocinética , Angelica sinensis , Animales , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/análisis , Extractos Vegetales/sangre , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem/métodos , Distribución TisularRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Blood deficiency is commonly encountered among women, and is the root of many gynecological disorders. Danggui Buxue Decoction (DBD), a classical traditional Chinese formula which is composed of Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) at the ratio of 5:1 (w/w), is widely used in TCM clinics for treatment of blood deficiency syndrome. This study is to compare the in vivo pharmacokinetic properties of seven major bio-active components in normal and blood deficiency rats after oral administration of DBD. MATERIALS AND METHODS: Blood deficiency rats were induced by bleeding from orbit at the dosages of 5.0mL/kg each day for 12 days. Normal and blood deficiency rats were administrated of DBD on the 12th day at the dosage of 20g/kg, and blood was collected at different time points after then. Concentrations of ferulic acid, caffeic acid, butylphthalide, ligustilide, calycosin-7-O-ß-glucoside, ononin, and astragaloside IV in plasma were quantified by UPLC-TQ/MS, and the main pharmacokinetic parameters were calculated by DAS 2.0. RESULTS: It was found that Cmax, Tmax and MRT0~T of astragaloside IV, Cmax, T1/2Z, AUC0~T and MRT0~T of calycosin-7-O-ß-glucoside, T1/2Z and AUC0~T of ferulic acid, T1/2Z, AUC0~T and MRT0~T of ononin, and MRT0~T of ligustilide, butylphthalide, and caffeic acid in blood deficiency rats was significantly different (P<0.05) from normal rats. CONCLUSIONS: This study was the first report about pharmacokinetic investigation in blood deficiency animals which was conducted by bleeding. And the results demonstrated that the seven DBD constituents in normal and blood deficiency rats had obvious differences in some pharmacokinetic characteristics, suggesting that the rate and extent of drug metabolism were altered in blood deficiency animals.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/farmacocinética , Enfermedades Hematológicas/metabolismo , Espectrometría de Masas/métodos , Animales , Área Bajo la Curva , Modelos Animales de Enfermedad , Femenino , Masculino , Medicina Tradicional China , Ratas , Ratas Sprague-DawleyRESUMEN
To explore rapidly the potential chemical markers for differentiating Radix Paeoniae Alba and Radix Paeoniae Rubra, a method is proposed based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) coupled with multivariate statistical analysis. Batches of commercial samples were analyzed by UPLC-Q-TOF/MS. The datasets of t(R)-m/z pair, ion intensities and sample codes were further processed with orthogonal partial least squared discriminant analysis (OPLS-DA) to compare holistically the difference between these two kinds of samples. Then statistics were used to generate an S-plot, in which the variables (t(R)-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components correlated to these ions should be regarded as the chemical markers. The identities of the most changed markers can be identified by comparing the mass/UV spectra and retention times with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of known compounds published in the literature. Using this proposed approach, albflorin, paeoniflorin, oxypaeoniflorin, benzoylpaeoniflorin, galloylalbiflorin and paeoniflorigenone were found to be the differentiating components for discrimination of Radix Paeoniae Alba and Radix Paeoniae Rubra. Moreover, paeoniflorin sulfonate and its isomer, isomaltopaeoniflorin sulfonate, were found to be the characteristic markers for all Radix Paeoniae Alba samples that were processed by sulfurdioxide gas fumigation. The results suggested that this newly established approach could be used to explore rapidly the potential chemical markers for herbs with similar chemical characteristics.
Asunto(s)
Paeonia/química , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Análisis MultivarianteRESUMEN
OBJECTIVE: To study the major metabolites of ferulic acid and gallic acid compatible with Danggui Chishaoyao in rat plasma and urine. METHOD: The metabolites of ferulic acid and gallic acid in rat plasma and urine were analyzed after oral administration of compatible Danggui Chishaoyao using UPLC-Q-TOF-MS. RESULT: On the basis of the mass information, it was inferred that in vivo metabolites of ferulic acid were be in the form of methylation products, sulfate conjugation products and glucuronidation conjugation products and so on; meanwhile, gallic acid was mainly transformed into eduction products and methylation products. CONCLUSION: There are kinds of phase I and phase II metabolites of ferulic acid and gallic acid in rat plasma and urine, which provide a basis for its efficacious materials and action mechanism.