ABSTRACT
Due to the limited resource of bear bile powder, the major raw material of Tanreqing Capsules(TRQ), cultured bear bile powder is used as a replacement to develop the Tanreqing Capsules Substitute(TRQS). An LC-MS/MS method was established in this study for simultaneous quantitation of 8 compounds from TRQS in rat plasma: tauroursodeoxycholic acid(TUDCA), taurocheno-deoxycholic acid(TCDCA), ursodeoxycholic acid(UDCA), chenodeoxycholic acid(CDCA), ferulic acid, wogonoside, baicalin, and forsythoside A. Thereby, the pharmacokinetic behaviors of TRQ and TRQS were evaluated. Concentration of endogenous compounds TUDCA, TCDCA, UDCA, and CDCA was determined with the stable isotope surrogate analytes: D4-TUDCA, D4-TCDCA, D4-UDCA, and D4-CDCA. Plasma samples were extracted by acetonitrile-induced protein precipitation. The LC conditions are as follows: Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 µm), mobile phase of 10 mmol·L~(-1) ammonium formate aqueous solution(containing 0.01% formic acid) and acetonitrile-methanol mixture(1â¶5). MS conditions are as below: multiple reaction monitoring(MRM), ESI~(+/-). Concentration of UDCA, CDCA, TUDCA, and TCDCA was corrected with a response factor, which is the ratio between the responses recorded for the surrogate and the authentic analyte at the equal concentration. Each of the plasma components showed good linearity(r > 0.995 1). Accuracy and precision met the criteria(inter-day RSD<7.0%, RE 89.98%-112.0%; intra-day RSD<12%, RE 90.41%-111.2%). The recovery was 64.83%-119.9% and matrix effect was 87.15%-113.8%. The validated method was applied for pharmacokinetic study of TRQS and TRQ(po, 0.94 g·kg~(-1)). There was no significant difference in C_(max) and AUC_(0-24 h) of baicalin, UDCA, TUDCA, and TCDCA between the two groups, indicating similar pharmacokinetic behaviors between TRQS and TRQ in rats.
Subject(s)
Drugs, Chinese Herbal/pharmacokinetics , Animals , Capsules , Chromatography, Liquid , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Tandem Mass SpectrometryABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Intrahepatic cholestasis is a common condition of many liver diseases with few therapies. Yinchenzhufu decoction (YCZFD) is a representative traditional Chinese herbal formula used for treating jaundice and liver disease. AIM OF THE STUDY: To investigate the hepatoprotective effect of YCZFD against cholestatic liver injury and reveal its potential mechanism. MATERIALS AND METHODS: Mice with alpha-naphthyl isothiocyanate (ANIT)-induced intrahepatic cholestasis were orally administered YCZFD at doses of 3, 6, and 12g crude drug/kg for 2 weeks followed by subsequent analyses. A serum metabolomics study was then performed to explore the different metabolites influenced by YCZFD using ultra-high-performance liquid chromatography coupled with linear ion trap-Orbitrap hybrid mass spectrometry (UPLC-LTQ-Orbitrap-MS/MS).The levels of individual bile acids in the serum, liver, and bile were determined by UPLC-MS/MS. The expression of metabolic enzymes, transporters, inflammatory factors, and cytokeratin-19 (CK-19) was determined by real-time PCR, western blotting, and immunohistochemistry. RESULTS: YCZFD administration decreased the serum biochemical indexes and ameliorated pathological damage, such as hepatic necrosis and inflammatory cell infiltration. Serum metabolomics revealed that the metabolites influenced by YCZFD were mainly associated with bile acid metabolism and inflammation. YCZFD administration effectively ameliorated the disordered bile acid homeostasis. The bile acid transporter, multidrug-resistance associated protein 2 (Mrp2), and the metabolic enzyme, cytochrome P450 2b10 (Cyp2b10), were upregulated in the YCZFD intervention group compared to those in the ANIT-induced group. YCZFD administration also significantly inhibited nuclear factor-κB (NF-κB) and its phosphorylation and decreased the expression of proinflammatory cytokines including tumor necrosis factor-α, interleukin-1ß, and intercellular adhesion molecule-1 in ANIT-induced cholestatic mice. Additionally, the level of CK-19 was lower in the YCZFD intervention group than in the ANIT-induced cholestatic mice. CONCLUSION: YCZFD administration ameliorated disordered bile acid homeostasis, inhibited NF-κB pathway-mediated inflammation, and protected the liver from bile duct injury. Therefore, YCZFD exerted a protective effect against cholestatic liver injury.
Subject(s)
Bile Acids and Salts/metabolism , Chemical and Drug Induced Liver Injury/prevention & control , Cholestasis, Intrahepatic/prevention & control , Drugs, Chinese Herbal/pharmacology , Homeostasis/drug effects , 1-Naphthylisothiocyanate , Animals , Bile/metabolism , Bile Acids and Salts/blood , Cholestasis, Intrahepatic/chemically induced , Cholestasis, Intrahepatic/metabolism , Dose-Response Relationship, Drug , Inflammation Mediators/blood , Keratin-19/blood , Male , Metabolomics , MiceABSTRACT
Intrahepatic cholestasis (IC) is a common syndrome that affects the liver, with treatment options being limited. Huangqi decoction (HQD), a classic herbal medicine, has shown protective effects against IC. In this study, isobaric tags for relative and absolute quantification-based quantitative proteomics was performed to investigate the potential mechanism of action of HQD on α-naphthylisothiocyanate (ANIT)-induced IC, resulting in 2796 quantified proteins across all samples, including 270 differentially expressed proteins under HQD treatment. Fuzzy c-means clustering analysis of these 270 proteins assigned the proinflammatory proteins, such as LCN2, SAA1, FGG, FGA, and FGB, to Cluster 1 (upregulated by ANIT, and downregulated by HQD). Functional bioinformatics and protein-protein interaction network analyses indicated that these proinflammatory proteins were involved in the STAT3 signaling pathway. Further real-time PCR and Western blot experiments confirmed that the expression of these proteins was consistent with the proteomic results. Moreover, HQD treatment decreased the phosphorylation of STAT3, induced by ANIT. Western blot experiments revealed that HQD treatment decreased phosphorylation of NF-κB and downregulated the expression of the inflammatory gene IL-6 and therefore inhibited the IL-6/STAT3 signaling pathway. In summary, the present study suggested that HQD may ameliorate acute cholestatic liver injury via inhibition of the NF-κB/IL-6/STAT3 signaling pathway.
Subject(s)
Interleukin-6 , NF-kappa B , Drugs, Chinese Herbal , Interleukin-6/genetics , Liver/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Proteomics , Signal TransductionABSTRACT
BACKGROUND: Huangqi decoction (HQD), a classic traditional herbal medicine, has been used for liver fibrosis, but its effect on intrahepatic chronic cholestatic liver injury remains unknown. PURPOSE: In the present study, we investigated the hepatoprotective effect of HQD and the underlying molecular mechanisms in 3, 5-diethoxycarbonyl-1, 4-dihydroxychollidine (DDC)-induced chronic cholestatic mice. METHODS: The DDC-induced cholestatic mice were administrated HQD for 4 or 8 weeks. Serum biochemistry and morphology were investigated. The serum and liver bile acid (BA) levels were detected by ultra performance liquid chromatography-tandem mass spectrometry. The liver expression of BA metabolizing enzymes and transporters, and inflammatory and fibrotic markers was measured by real-time polymerase chain reaction, western blotting, and immunohistochemistry. RESULTS: HQD treatment for 4 or 8 weeks ameliorated DDC-induced liver injury by improving impaired hepatic function and tissue damage. HQD treatment for 8 weeks further decreased the liver expression of cytokeratin 19, tumor growth factor (TGF)-ß, collagen I, and α-smooth muscle actin, and ameliorated ductular reaction and liver fibrosis. HQD markedly decreased the accumulation of serum and liver BA. The expression of BA-metabolizing enzymes, cytochrome P450 2b10 and UDP glucuronosyltransferase 1 A1, and multidrug resistance-associated protein 2, Mrp3, and Mrp4 involved in BA homeostasis was increased by 4 weeks of HQD treatment. The expression of BA uptake transporter Na+-taurocholate cotransporting polypeptide was decreased and that of Mrp4 was increased after 8 weeks of HQD treatment. Nuclear factor-E2-related factor-2 (Nrf2) was remarkably induced by HQD treatment. Additionally, HQD treatment for 8 weeks decreased the liver expression of inflammatory factors, interleukin (IL)-6, IL-1ß, tumor necrosis factor-α, monocyte chemoattractant protein-1, and intracellular adhesion molecule-1. HQD suppressed the nuclear factor (NF)-κB pathway. CONCLUSION: HQD protected mice against chronic cholestatic liver injury and biliary fibrosis, which may be associated with the induction of the Nrf2 pathway and inhibition of the NF-κB pathway, ameliorating BA-stimulated inflammation.
Subject(s)
Bile Acids and Salts/metabolism , Cholestasis, Intrahepatic/drug therapy , Drugs, Chinese Herbal/pharmacology , Animals , Cholestasis, Intrahepatic/chemically induced , Cholestasis, Intrahepatic/metabolism , Cholestasis, Intrahepatic/pathology , Dicarbethoxydihydrocollidine , Drugs, Chinese Herbal/chemistry , Enzymes/metabolism , Hepatitis/drug therapy , Hepatitis/etiology , Liver/drug effects , Liver/metabolism , Liver/pathology , Liver Cirrhosis/drug therapy , Liver Cirrhosis/pathology , Male , Mice, Inbred C57BL , NF-E2-Related Factor 2/metabolism , NF-kappa B/antagonists & inhibitors , NF-kappa B/metabolism , Protective Agents/pharmacologyABSTRACT
Pharmacokinetic studies are crucial for elucidating the effective constituents and formula compatibility of traditional Chinese medicines (TCMs). However, studies have usually been limited to single dosages and detection of systemic blood concentrations. To obtain comprehensive pharmacokinetic information, here we propose a multi-dosage and multi-sampling (blood from portal vein or systemic circulation, and liver) strategy to comparatively study the pharmacokinetics of multi-form TCMs, i.e., pure constituents, TCMs, or TCM formula extracts. Based on this strategy, we studied the pharmacokinetics of pure berberine, berberine in CoptidisRhizoma (CRE), and berberine in CoptidisRhizoma-GlycyrrhizaeRadix etRhizoma extracts (CR-GRE). After simple calculation and comparison of the obtained area under the curve (AUC) values, the results revealed the drastically different pharmacokinetic properties of pure berberine compared to CRE and CR-GRE. The results contribute to explaining the pharmacological loss of berberine activity after purification and the compatibility of the CR-GR drug pair. The results also innovatively showed that it was intestinal absorption that differentiated the pharmacokinetics of CRE and pure berberine, and CRE and CR-GRE. In conclusion, we propose a composite strategy to comparatively study the pharmacokinetics of TCMs, which could provide sufficient information to obtain a comprehensive view, before follow-up mechanism-of-action studies.
Subject(s)
Berberine/chemistry , Berberine/pharmacokinetics , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Animals , Antioxidants/chemistry , Antioxidants/pharmacokinetics , Area Under Curve , Chemistry, Pharmaceutical , Dose-Response Relationship, Drug , Female , Humans , Intestinal Absorption/physiology , Male , Medicine, Chinese Traditional , Mice , Molecular Structure , Terpenes/pharmacologyABSTRACT
A rapid, sensitive and accurate UPLC-MS/MS method was developed for the simultaneous quantification of components of Huangqi decoction (HQD), such as calycosin-7-O-ß-d-glucoside, calycosin-glucuronide, liquiritin, formononetin-glucuronide, isoliquiritin, liquiritigenin, ononin, calycosin, isoliquiritigenin, formononetin, glycyrrhizic acid, astragaloside IV, cycloastragenol, and glycyrrhetinic acid, in rat plasma. After plasma samples were extracted by protein precipitation, chromatographic separation was performed with a C18 column, using a gradient of methanol and 0.05% acetic acid containing 4mm ammonium acetate as the mobile phase. Multiple reaction monitoring scanning was performed to quantify the analytes, and the electrospray ion source polarity was switched between positive and negative modes in a single run of 10 min. Method validation showed that specificity, linearity, accuracy, precision, extraction recovery, matrix effect and stability for 14 components met the requirements for their quantitation in biological samples. The established method was successfully applied to the pharmacokinetic study of multiple components in rats after intragastric administration of HQD. The results clarified the pharmacokinetic characteristics of multiple components found in HQD. This research provides useful information for understanding the relation between the chemical components of HQD and their therapeutic effects.
Subject(s)
Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/pharmacokinetics , Tandem Mass Spectrometry/methods , Administration, Oral , Animals , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/chemistry , Female , Flavonoids/blood , Flavonoids/chemistry , Flavonoids/pharmacokinetics , Glucosides/blood , Glucosides/chemistry , Glucosides/pharmacokinetics , Glycosides/blood , Glycosides/chemistry , Glycosides/pharmacokinetics , Linear Models , Male , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and Specificity , Terpenes/blood , Terpenes/chemistry , Terpenes/pharmacokineticsABSTRACT
Intrahepatic cholestasis is a serious symptom of liver disorders with limited therapies. In this study, we investigated the efficacy of Huangqi decoction (HQD), a two-herb classic traditional Chinese medicine (TCM), in the treatment of alpha-naphthylisothiocyanate (ANIT)-induced intrahepatic cholestasis in mice. HQD treatment ameliorated impaired hepatic function and tissue damage. A metabolomics study revealed that the endogenous metabolites significantly affected by HQD were related to bile acid (BA) biosynthesis and glutathione metabolism pathways. HQD treatment decreased the intrahepatic accumulation of cytotoxic BAs, normalized serum BA levels, and increased biliary and urinary BA excretion. Additionally, HQD restored the hepatic glutathione content and suppressed reactive oxygen species (ROS) in cholestatic mice. Protein and gene analysis revealed that HQD increased the expression of the hepatic metabolizing enzymes cytochrome P450 (CYP) 2B10 and UDP glucuronosyltransferase family 1 member A1 (UGT1A1), as well as multidrug resistance-associated protein 2 (Mrp2), Mrp3, and Mrp4, which play crucial roles in BA homeostasis. Further, HQD increased the protein expression of glutamate-cysteine ligase, which is involved in the synthesis of glutathione. Importantly, HQD increased the nuclear expression of nuclear factor-E2-related factor-2 (Nrf2). In conclusion, HQD protects against intrahepatic cholestasis by reversing the disordered homeostasis of BAs and glutathione.
ABSTRACT
Traditional Chinese medicine (TCM) has a long history of medical use in China and is still used worldwide. Unexpected herb-drug interactions (HDIs) may lead to adverse drug reactions or loss of therapeutic efficacy of the victim drug. Here, based on searches of Medline, EBSCO, Science Direct and Web of Science using various keywords, we summarize the TCM-derived pharmacokinetic HDIs that were reported from 1990 to 2015 and discuss the underlying mechanisms. In general, many pre-clinical and clinical pharmacokinetic HDIs have been reported. Our searches show that TCMs cause pharmacokinetic interactions with therapeutic drugs mainly by inhibiting or inducing drug-metabolizing enzymes and transporters. However, most of the interactions result from a small number of prescription medications and the actual potential for harm is low. Moreover, such HDIs can be avoided by discontinuing the TCMs. Despite the extensive number of reports on TCM-derived HDIs, the findings are frequently conflicting and can be confusing. The causes of the conflicts vary, but we classified them into three basic categories as follows: (1) complicated nature and poor quality control of TCMs, (2) different responses of various test systems to TCM exposure and (3) diverse study designs. Accordingly, we propose rational study designs for future HDI research. We also propose that a specific authoritative guide be established that provides recommendations for HDI studies. This review provides insights into the progress and challenges in TCM-derived pharmacokinetic HDI research.
Subject(s)
Drugs, Chinese Herbal/pharmacokinetics , Herb-Drug Interactions , Animals , Drugs, Chinese Herbal/pharmacology , Humans , Medicine, Chinese Traditional/adverse effects , Medicine, Chinese Traditional/methodsABSTRACT
Pharmacological activities of some natural products diminish and even disappear after purification. In this study, we explored the mechanisms underlying the decrease of acute oral toxicity of Coptidis Rhizoma extract after purification. The water solubility, in vitro absorption, and plasma exposure of berberine (the major active compound) in the Coptidis Rhizoma extract were much better than those of pure berberine. Scanning electron microscopy, laser scanning confocal microscopy (LSCM), and dynamic light scattering experiments confirmed that nanoparticles attached to very fine precipitates existed in the aqueous extract solution. The LSCM experiment showed that the precipitates were absorbed with the particles by the mouse intestine. High-speed centrifugation of the extract could not remove the nanoparticles and did not influence plasma exposure or acute oral toxicity. However, after extract dilution, the attached precipitates vanished, although the nanoparticles were preserved, and there were no differences in the acute oral toxicity and plasma exposure between the extract and pure berberine. The nanoparticles were then purified and identified as proteinaceous. Furthermore, they could absorb co-dissolved berberine. Our results indicate that naturally occurring proteinaceous nanoparticles in Coptidis Rhizoma extract act as concentration-dependent carriers that facilitate berberine absorption. These findings should inspire related studies in other natural products.
Subject(s)
Berberine , Drugs, Chinese Herbal , Nanoparticles , Plant Proteins , Animals , Berberine/pharmacokinetics , Berberine/toxicity , Coptis chinensis , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/toxicity , Mice , Nanoparticles/chemistry , Nanoparticles/toxicity , Plant Proteins/chemistry , Plant Proteins/toxicityABSTRACT
Entecavir (ETV), a guanosine nucleotide antiviral agent with activity against hepatitis B virus (HBV) and Huangqi decoction (HQD) that exerts significant therapeutic effects in liver cirrhosis are used as an effective drug combination in the treatment of liver cirrhosis with HBV. Therefore, this study was designed to assess the effect of HQD on ETV pharmacokinetics in rat plasma. Spraque-Dawley (SD) rats were randomized into single- and 7-day-dose experimental groups. The ETV and ETV-HQD groups were administered ETV and a simultaneous combination of ETV and HQD, respectively while the ETV-HQD-2h group received HQD 2 h after ETV treatment, all administered via intragastric (i.g.) gavage. A rapid, sensitive, and efficient ultra-high- performance liquid chromatography-linear trap quadrupole (UHPLC-LTQ)-Orbitrap method was developed and validated to determine ETV in rat plasma from blood samples collected at different time points following treatment. The linearity, accuracy, precision, recovery, matrix effects and stability of ETV were all satisfactory. The ETV-HQD group exhibited a decrease in the maximum plasma concentration (Cmax), and a delay in time to achieve Cmax (tmax) following single- and multi-dose administrations, and decreased area under the concentration- time curve (AUC0t) following single dosing. ETV pharmacokinetics did not change significantly between the ETV and ETV-HQD-2h groups. In vitro everted intestinal sac models experiments indicated that HQD decreased the absorption of ETV. HQD prevented ETV from accessing the intestinal mucosa epithelial surface, thereby decreasing its absorption in rats.
Subject(s)
Antiviral Agents/pharmacokinetics , Drugs, Chinese Herbal/pharmacology , Guanine/analogs & derivatives , Adult , Animals , Antiviral Agents/blood , Chromatography, High Pressure Liquid , Guanine/blood , Guanine/pharmacokinetics , Humans , Plasma , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
Xiexin decoction, a herbal therapeutic agent commonly used in traditional Chinese medicine, is recognized for its beneficial effects on diabetic nephropathy exerted through the combined action of multiple components, including Rhizoma Coptidis alkaloids (A), Radix et Rhizoma Rhei polysaccharides (P), and Radix Scutellaria flavones (F). Our previous studies have shown that a combination of A, P, and F (APF) exhibits renoprotective effects against diabetic nephropathy. This study was aimed at determining the effects of APF on renal fibrosis in diabetic nephropathy and elucidating the underlying molecular mechanisms. To evaluate the effects of APF, in vivo, db/db diabetic mice were orally administered a low or high dose of APF (300 or 600 mg/kg, respectively) once a day for 8 weeks. We evaluated the blood and urine indices of metabolic and renal function, renal tissue histopathology, renal inflammation, and fibrosis. APF treatment significantly ameliorated glucose and lipid metabolism dysfunction, decreased urinary albumin excretion, normalized creatinine clearance, and reduced the morphological changes in renal tissue. Additionally, APF administration in db/db diabetic mice reduced the elevated levels of renal inflammation mediators such as intercellular adhesion molecule-1, monocyte chemotactic protein-1, tumor necrosis factor-α, interleukin-1ß, and active nuclear factor κB (NF-κB). APF treatment also reduced type I and IV collagen, transforming growth factor-ß1 (TGF-ß1), and TGF-ß1 type II receptor expression levels, and decreased the phosphorylation of Smad2/3 in the kidneys of db/db diabetic mice. These results suggest that APF reduces renal fibrosis in diabetic nephropathy through the NF-κB and TGF-ß1/Smad signaling pathways. In vitro, APF treatment reduced cell proliferation and protein expression of α-smooth muscle actin, collagen I, TGF-ß1 and NF-κB in mesangial cells cultured with high glucose concentrations. Our findings indicate that treatment with multi-component herbal therapeutic formulations may be a useful approach for the treatment of diabetic nephropathy.
Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Kidney Diseases/drug therapy , NF-kappa B/antagonists & inhibitors , Signal Transduction/drug effects , Analysis of Variance , Animals , Blotting, Western , DNA Primers/genetics , Fibrosis , Immunohistochemistry , Mice , Mice, Mutant Strains , Microscopy, Electron , Real-Time Polymerase Chain Reaction , Smad Proteins/antagonists & inhibitors , Transforming Growth Factor beta1/antagonists & inhibitorsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Xiexin decoction (XXD) has been used as a treatment for diabetes mellitus for more than 1300 years. XXD constituents with protective effects against diabetic nephropathy (DN) include Rhizoma Coptidis alkaloids (RA), Radix et Rhizoma Rhei polysaccharides (RP), and Radix Scutellaria flavones (RF). The aim of the study is to investigate the effects of combinations of RA, RP, and RF on DN and their mechanisms of action. MATERIALS AND METHODS: In vitro, high glucose-induced rat mesangial cells were treated with RA, RP, RF, and combinations thereof. Cell proliferation and levels of inflammatory factors were measured. In vivo, high-fat diet and streptozotocin-induced diabetic rats were treated with different combinations of RA, RP, and RF once per day for 12 weeks. Blood and urine biochemical parameters, renal tissue morphology, and inflammation were investigated. RESULTS: In vitro, the combination of the three groups of components inhibited mesangial cell proliferation and reduced the levels of monocyte chemotactic protein-1 (MCP-1) and collagen IV. The effects of the three constituent groups in combination were stronger than those of each group alone or combinations of two groups. In diabetic rats, combinations of the three groups of herb components ameliorated blood glucose, urinary albumin excretion and decreased renal mesangial matrix expansion and basement membrane thickening. In addition, the combinations reduced renal tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) protein levels, down-regulated the expression of nuclear factor κB (NF-κB) and transforming growth factor beta 1 (TGF-ß1), and up-regulated the expression of inhibitor of nuclear factor κB (IκB) protein. Among the three groups of herb components, RA produced the strongest effects, followed by RP, and then by RF. CONCLUSIONS: The combination of the three groups of herb components produced anti-DN effects through inhibition of inflammation mediated by NF-κB. Among the three groups of herb components, RA produced the strongest effect while RP and RF produced weaker effects.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Diabetic Nephropathies/drug therapy , Drugs, Chinese Herbal/pharmacology , Inflammation/drug therapy , Animals , Cell Proliferation/drug effects , Diabetes Mellitus, Experimental/complications , Diabetic Nephropathies/pathology , Diet, High-Fat , Drugs, Chinese Herbal/chemistry , Gene Expression Regulation/drug effects , Male , Mesangial Cells/drug effects , Mesangial Cells/metabolism , NF-kappa B/metabolism , Rats , Rats, Sprague-Dawley , StreptozocinABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Yinchenzhufu decoction (YCZFD) is a classical Chinese herbal formula and has been used to treat severe jaundice in chronic liver injuries since the Qing Dynasty (18th century CE). To identify the components absorbed into the blood in YCZFD and explore their pharmacokinetic profile for understanding the effective ingredients of YCZFD. MATERIALS AND METHODS: After rats were given YCZFD by intragastric administration, the plasma was processed by precipitation of protein. The compounds in YCZFD extract and the plasma were identified by using high-resolution mass spectrometry with a database-directed strategy. The pharmacokinetics of multiple compounds from YCZFD in rat plasma was studied by using the established UPLC-MS/MS method. RESULTS: Forty compounds in YCZFD extract and 21 prototype compounds with 11 metabolites in rat plasma were detected after oral administration. The pharmacokinetic parameters of glycyrrhizic acid, glycyrrhetic acid, cinnamic acid, ononin, atractylenolide III, and liquiritin from YCZFD were obtained in rats. CONCLUSIONS: The identified constituents and the pharmacokinetic features of YCZFD are helpful for understanding the material bases of its therapeutic effects.
Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Animals , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/analysis , Female , Male , Phytochemicals/isolation & purification , Phytochemicals/pharmacokinetics , Rats, Sprague-Dawley , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
Fuzi Xiexin Tang (FXT) is a classic traditional Chinese medicine formula which has been employed in clinical for more than 1800 years. The distinctive preparation method (maceration) recorded in ancient time is different from one in modern clinical practice (decoction). Aim of this study is to investigate the pharmacokinetic difference of alkaloids, flavones and anthraquinones in rats after oral administration of decoction of FXT (DFXT, 30gkg(-1)), maceration of FXT (MFXT, 30gkg(-1)) and decoction of Aconiti Lateralis Radix Preparata (DAR, 6gkg(-1)) by a validated UPLC-MS/MS method. Plasma concentrations and pharmacokinetic parameters of 16 active constituents (aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, berberine, palmatine, jatrorrhizine, coptisine, baicalin, wogonin, wogonoside, emodin, aloe-emodin, rhein) in rat were quantified and compared. Different preparative methods resulted in significant difference on exposure and pharmacokinetic characteristics of alkaloids, flavones and anthraquinones from FXT, especially protoberberine alkaloids. Concentrations of monoester-diterpenoid alkaloids were below the LOD in rat plasma after administration of DFXT and MFXT because of the existence of other three herbs from FXT. Maceration could decrease the absorption of flavones while increased the absorption of anthraquinones. Cmax of emodin and rhein were 3.1 and 10.3 times increased, while eliminations of these two constituents were 8.0 and 19.0 times slower after administration of MFXT. Bioavailability of both flavones and anthraquinones increased after administration of MFXT, especially emodin and rhein increasing as much as 13.5 and 20.7 times. Herb-herb interaction between DAR and other three herbs from FXT significantly influenced the exposure of aconitum alkaloids.
Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Plant Extracts/chemistry , Plant Extracts/pharmacokinetics , Absorption , Administration, Oral , Alkaloids/chemistry , Alkaloids/pharmacokinetics , Animals , Anthraquinones/chemistry , Anthraquinones/pharmacokinetics , Biological Availability , Chromatography, High Pressure Liquid/methods , Diterpenes , Flavones/chemistry , Flavones/pharmacokinetics , Medicine, Chinese Traditional/methods , Rats , Rats, Sprague-Dawley , Tandem Mass Spectrometry/methodsABSTRACT
In this paper, absorption and pharmacokinetic study of Radix Rehmanniae was studied by liquid chromatography coupled with mass spectrometry method after oral administration to rats. By comparing the chromatograms of ultraviolet, full scan, extracted ion and selective reaction monitoring (SRM) of standard solution, Radix Rehmanniae, blank plasma and rat plasma post drug administration, catalpol and ajugol were found to be the main compounds absorbed from Radix Rehmanniae. Plasma concentrations of aucubin, dihydrocatalpol, rehmannioside A (or rehmannioside B/ melittoside) and rehmannioside D were very low. Quantitative method for catalpol and aucubin and semi-quantitative method for other compounds in rat plasma were established. The pharmacokinetic study of those absorbed components was conducted after oral administration of 6 g x kg(-1) Radix Rehmanniae water extract to rats. Cmax, t(1/2) and AUC(0-infinity) of catalpol and ajugol were (2349.05 +/- 1438.34) and (104.25 +/- 82.05) ng x mL(-1), (0.86 +/- 0.32) and (0.96 +/- 0.37) h, (4407.58 +/- 2734.89) and (226.66 +/- 188.38) ng x h x mL(-1), respectively. tmax was at 1.00 h for catalpol and ajugol. Both catalpol and ajugol were absorbed and excreted rapidly.
Subject(s)
Drugs, Chinese Herbal/pharmacokinetics , Iridoid Glucosides/pharmacokinetics , Iridoid Glycosides/pharmacokinetics , Pyrans/pharmacokinetics , Rehmannia/chemistry , Administration, Oral , Animals , Area Under Curve , Drugs, Chinese Herbal/chemistry , Female , Iridoid Glucosides/blood , Iridoid Glucosides/chemistry , Iridoid Glycosides/blood , Iridoid Glycosides/chemistry , Male , Molecular Structure , Plant Roots/chemistry , Plants, Medicinal/chemistry , Pyrans/blood , Pyrans/chemistry , Rats , Rats, Sprague-DawleyABSTRACT
The pharmacokinetics and anti-inflammatory effects of Sanhuang Xiexin Tang, composed of Rhei Radix et Rhizoma, Scutellariae Radix, and Coptidis Rhizoma, prepared by decoction and maceration, were investigated and compared. Rats were orally administered with the decoction and maceration of Sanhuang Xiexin Tang at 30 g/kg. The concentrations of 10 active constituents (berberine, palmatine, jatrorrhizine, coptisine, wogonin, baicalin, wogonoside, emodin, aloe-emodin, and rhein) in plasma were determined by UPLC-MS/MS. Mice were orally administered decoctions and macerations of Sanhuang Xiexin Tang continuously for 7 days at three doses and stimulated with lipopolysaccharide. The plasma concentrations of IL-10 and TNF-α were determined by ELISA. Different preparation methods resulted in significant differences in the pharmacokinetic characteristics of the SXT constituents, especially the protoberberine alkaloids. Maceration decreased the absorption of flavones while promoting the absorption of anthraquinones. Bioavailability of both flavones and anthraquinones increased after administration of macerated Sanhuang Xiexin Tang, especially those of baicalin and rhein, which increased by 3.27 and 7.10 times. Results of ELISA indicated that both the decoction and maceration of Sanhuang Xiexin Tang could significantly increase IL-10 production (p < 0.01) as well as decrease TNF-α production (p < 0.01). Macerated Sanhuang Xiexin Tang has a slightly higher anti-inflammatory effect than the Sanhuang Xiexin Tang decoction. Different preparation methods affected the pharmacokinetic characteristics and anti-inflammatory effects of Sanhuang Xiexin Tang's active constituents.
Subject(s)
Alkaloids/pharmacology , Anthraquinones/pharmacology , Anti-Inflammatory Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Flavonoids/pharmacology , Inflammation/blood , Magnoliopsida/chemistry , Alkaloids/pharmacokinetics , Animals , Anthraquinones/pharmacokinetics , Anti-Inflammatory Agents/pharmacokinetics , Coptis/chemistry , Cytokines/blood , Drugs, Chinese Herbal/metabolism , Drugs, Chinese Herbal/pharmacokinetics , Flavones/pharmacokinetics , Flavones/pharmacology , Flavonoids/pharmacokinetics , Interleukin-10/blood , Mice , Mice, Inbred Strains , Rats , Rats, Sprague-Dawley , Rheum/chemistry , Scutellaria/chemistry , Tumor Necrosis Factor-alpha/bloodABSTRACT
In Chinese medicine, Xiexin decoction (XXD) has been used for the clinical treatment of diabetes for at least 1700 years. The present study was conducted to investigate the effective ingredients of XXD and their molecular mechanisms of antidiabetic nephropathy in rats. Rats with diabetes induced by high-fat diet and streptozotocin were treated with XXD extract for 12 weeks. XXD significantly improved the glucolipid metabolism disorder, attenuated albuminuria and renal pathological changes, reduced renal advanced glycation end-products, inhibited receptor for advanced glycation end-product and inflammation factors expression, suppressed renal nuclear factor- κ B pathway activity, and downregulated renal transforming growth factor- ß 1. The concentrations of multiple components in plasma from XXD were determined by liquid chromatography and tandem mass spectrometry. Pharmacokinetic/pharmacodynamic analysis using partial least square regression revealed that 8 ingredients of XXD were responsible for renal protective effects via actions on multiple molecular targets. Our study suggests that the renal protective role of XXD with multiple effective ingredients involves inhibition of inflammation through downregulation of the nuclear factor- κ B pathway, reducing renal advanced glycation end-products and receptor for advanced glycation end-product in diabetic rats.
ABSTRACT
A rapid and sensitive UPLC-ESI/MS method was established and validated to determine 17 active constituents (aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, berberine, palmatine, jatrorrhizine, coptisine, baicalein, wogonin, baicalin, wogonoside, emodin, aloe-emodin and rhein) in Sanhuang Xiexin Tang (SXT) and Fuzi Xiexin Tang (FXT), which are two classic compound recipes from Xiexin Tang categorized formulas in traditional Chinese medicien. The separation was performed on a UPLC BEH C18 column gradient eluted using acetonitrile and 0.1% formic acid as mobile phase. ESI/MS was operated in positive ([M + H](+)) in selected ion recording mode for analysis of alkaloids and flavones, while in negative ([M - H](-)) selected ion recording mode for anthraquinones. All of the 17 constituents exhibited good linearity in a relatively wide concentration ranges with the lowest limits of detection of 0.38 ng/mL. All of the relative standard deviation values of intra- and inter-precisions and stabilities of 17 constituents were within 5%. The method was successfully applied to determine 17 active constituents in decoctions and macerations of SXT and FXT. The results indicated that different preparative methods resulted in significant diversity in concentrations of the 17 analytes. Herb-herb interaction appeared between aconitum alkaloids in Aconiti Lateralis Radix Preparata and another three herbs.
Subject(s)
Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/analysis , Drugs, Chinese Herbal/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Alkaloids/analysis , Alkaloids/chemistry , Alkaloids/isolation & purification , Anthraquinones/analysis , Anthraquinones/chemistry , Anthraquinones/isolation & purification , Drug Stability , Flavonoids/analysis , Flavonoids/chemistry , Flavonoids/isolation & purification , Limit of Detection , Linear Models , Reproducibility of ResultsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Xie-xin decoction (XXD) has been used as a classic formula in China for the treatment of gastrointestinal dysfunction such as ulcerative colitis (UC). However, no potential action mechanisms and active compounds had been systematically investigated. AIM OF THE STUDY: To explore the effectiveness and the material basis of XXD in trinitrobenzene sulfonic acid (TNBS)-induced UC rats. MATERIALS AND METHODS: XXD was administered orally for 8 days at a dosage of 2 or 4g/kg/day. Plasma pharmacokinetic properties and colon tissue concentrations of multiple compounds from XXD were detected. Tissue damage scores, production of interleukin (IL)-10 and myeloperoxidase (MPO), expression of tumor necrosis factor-alpha (TNF-α) and nuclear factor-kappa Bp65 (NF-κBp65) in colon tissues were examined. Canonical correlation analysis was performed to evaluate the relationships between pharmacokinetics and efficacy to elucidate significantly active compounds of XXD. RESULTS: XXD promoted the recovery of colitis and inhibited the colonic inflammation damage in UC rats by reducing the level of MPO and the expression of TNF-α and NF-κBp65, and increasing the production of IL-10 in colon tissues. Efficacy of XXD was positively related with AUC of five plasma compounds (baicalin, berberine, wogonoside, wogonin, and rhein) and concentrations of six colon tissue compounds (coptisine, jatrorrhizine, palmatine, berberine, baicalein and emodin), respectively. CONCLUSIONS: The multiple compounds in plasma and colon tissues from XXD might be the main material basis for therapeutic potentials in UC rats.
Subject(s)
Colitis, Ulcerative/drug therapy , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/therapeutic use , Animals , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/metabolism , Colitis, Ulcerative/pathology , Interleukin-10/metabolism , Male , Peroxidase/metabolism , Phytotherapy , Rats , Rats, Sprague-Dawley , Transcription Factor RelA , Treatment Outcome , Trinitrobenzenesulfonic Acid , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Fugan fang (FGF) is a traditional Chinese medicine (TCM) prescription that has been used in treating hepatic illnesses for many years. In this study, an analytical method is developed for the quantitative analysis of the major components of FGF. This method is based on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS) on a reverse-phase C18 column. Results show that 0.01% acetic acid and acetonitrile is the optimum mobile phase in gradient elution. All compounds showed good linearity (r(2) ≥ 0.9948). Recoveries measured at three concentration levels varied from 83.5 to 104.8%. The method was validated with respect to precision, repeatability and accuracy and was successfully applied in the quantification of the 11 components of FGF products. The validated HPLC-MS method provides a basis for assessing the quality of TCM prescriptions containing many bioactive components.