ABSTRACT
Stroke is a multifactorial disease and a consequence of morbidities of diabetes, obesity, hypertension, and heart diseases. Leptin is a major adipokine that regulates weight balance and energy homeostasis, the level of which has been considered as an indicator of acute ischemic stroke. In the present study, we confirmed the high level of leptin and noradrenaline in stroke patients and mouse models as well as oxygen-glucose deprivation (OGD) primary cerebral neurons. Leptin administration increased noradrenaline concentration and dopamine ß-monooxygenase (DBH) but decreased noradrenaline transporter (NET) expression in primary cerebral neurons. Moreover, induced noradrenaline concentration, DBH activity, and inhibited NET were blunted by TG101348 (JAK2 inhibitor). JAK2 silencing also abolished the effects of leptin on noradrenaline metabolism.
Subject(s)
Brain Ischemia/drug therapy , Brain Ischemia/metabolism , Leptin/pharmacology , Norepinephrine/metabolism , Animals , Case-Control Studies , Cells, Cultured , Dopamine beta-Hydroxylase/metabolism , Humans , Janus Kinase 2/antagonists & inhibitors , Janus Kinase 2/metabolism , Male , Mice , Neurons/cytology , Neurons/metabolism , Norepinephrine Plasma Membrane Transport Proteins/metabolism , Pyrrolidines/pharmacology , Sulfonamides/pharmacologyABSTRACT
Tanreqing injection (TRQ), a well-known traditional Chinese medicine formula, is commonly used to treat respiratory diseases. In the present study, a rapid, selective, and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to simultaneously determinate the plasma contents of 5 major constituents of TRQ, including chlorogenic acid (CHA), caffeic acid (CFA), baicalin (BA), ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) in rats after intravenous administration of TRQ. Chromatographic separation was performed on an Agilent Zorbax SB-C column (3.5 μm, 100 mm × 2.1 mm), with acetonitrile and 0.1% aqueous formic acid as mobile phase at a flow rate of 0.3 mL·min. The calibration curves were linear over the ranges of 27.0-13 333.0 ng·mL for CFA, 30.0-14 933.0 ng·mL for CHA, 50.0-50 333.0 ng·mL for BA, 550.0-55 000.0 ng·mL for UDCA, and 480.0-48 000.0 ng·mL for CDCA, respectively. Intra- and inter-day precisions (relative standard deviations, RSDs) were from 3.11% to 14.08%. The extraction recoveries were greater than 71% and accuracy (relative recovery) was from 89% to 137% for all analytes, except endogenous bile acids. This validated method was successfully applied to the first pharmacokinetic study of CFA, CHA, BA, UDCA and CDCA in rat plasma after intravenous administration of TRQ.
Subject(s)
Animals , Male , Rats , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Pharmacokinetics , Molecular Structure , Rats, Sprague-Dawley , Tandem Mass Spectrometry , MethodsABSTRACT
This study aimed to profile the chemical constituents of Zi-Shen pill (ZSP) and its metabolites in plasma, urine, and prostate tissue, after administration into rats. Based on the chromatographic retention behavior, fragmentation patterns of chemical components, published literatures, and literature databases, an UPLC-Q-TOF/MS (LC-TOF/MS) method was established to identify the components of ZSP and its metabolites in biological samples. A total of 101 compounds were identified and tentatively characterized from the ZSP, including alkaloids, xanthones, and timosaponins. Except for 33 prototype components, 22 metabolites were detected in the plasma, urine, and prostate, and mainly came from Phellodendri Amurensis Cortex and Anemarrhenae Rhizoma. It was found that glucuronidation and sulfation were the major metabolic processes of xanthones, while oxidation, demethylation, and glucuronidation were the major metabolic pathways of alkaloids. In summary, the present study provided important chemical information on the metabolism of ZSP, indicating that alkaloids might be able to be absorbed into the prostate. The results provided a basis for further studies of the mechanisms of action for ZSP.
Subject(s)
Animals , Male , Rats , Alkaloids , Blood , Urine , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Metabolism , Plasma , Chemistry , Tandem Mass Spectrometry , Urine , Chemistry , Xanthones , Blood , UrineABSTRACT
AIMS@#To develop an HPLC-MS/MS method for the quantification of platycodin D (PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract (PRE) containing PD.@*METHOD@#Plasma samples were pretreated with solid-phase extraction using an Oasis® HLB SPE cartridge. Madecassoside was used as the internal standard (IS). Chromatographic separation was achieved on an ODS column (100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water (30 : 70, V/V) containing 0.1 mmol·L(-1) ammonium acetate at a flow rate of 0.25 mL·min(-1). The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization (ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside (IS), respectively.@*RESULTS@#The calibration curve was linear from 5 to 2 000 ng·mL(-1) (r(2) >0.99) with a lower limit of quantification (LLOQ) of 5 ng·mL(-1). The intra- and inter-day precision (relative standard deviation, RSD) values were below 15% and the accuracy (relative error, RE) was from -15% to +15% at three quality control (QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be (0.48 ± 0.19)% when administered PD, and to be (1.81 ± 0.89)% when administered PRE.@*CONCLUSION@#The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.
Subject(s)
Animals , Male , Rats , Administration, Oral , Biological Availability , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Pharmacokinetics , Plant Roots , Chemistry , Platycodon , Chemistry , Rats, Sprague-Dawley , Saponins , Blood , Pharmacokinetics , Tandem Mass Spectrometry , Methods , Triterpenes , Blood , PharmacokineticsABSTRACT
AIM@#To develop and validate a high performance liquid chromatography (HPLC) coupled with diode array and evaporative light scattering detectors (DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes (namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair (ZB).@*METHOD@#Chromatographic separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min(-1) at 260 nm. The drift tube temperature of ELSD was set to 60 °C and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy.@*RESULTS@#The HPLC-DAD-ELSD method allowed the quantification of ten compounds (phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples.@*CONCLUSION@#This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.
