ABSTRACT
Ischemic brain stroke is pathologically characterized by tissue acidosis, sustained calcium entry and progressive cell death. Previous studies focusing on antagonizing N-methyl-d-aspartate (NMDA) receptors have failed to translate any clinical benefits, suggesting a non-NMDA mechanism involved in the sustained injury after stroke. Here, we report that inhibition of intracellular proton-sensitive Ca2+-permeable transient receptor potential vanilloid 3 (TRPV3) channel protects against cerebral ischemia/reperfusion (I/R) injury. TRPV3 expression is upregulated in mice subjected to cerebral I/R injury. Silencing of TRPV3 reduces intrinsic neuronal excitability, excitatory synaptic transmissions, and also attenuates cerebral I/R injury in mouse model of transient middle cerebral artery occlusion (tMCAO). Conversely, overexpressing or re-expressing TRPV3 increases neuronal excitability, excitatory synaptic transmissions and aggravates cerebral I/R injury. Furthermore, specific inhibition of TRPV3 by natural forsythoside B decreases neural excitability and attenuates cerebral I/R injury. Taken together, our findings for the first time reveal a causative role of neuronal TRPV3 channel in progressive cell death after stroke, and blocking overactive TRPV3 channel may provide therapeutic potential for ischemic brain injury.
ABSTRACT
Objective: To establish the Q-Marker database for Chinese medicine Qingreling Granules (QG) based on the concept of Q-Marker by the method of ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-quadrupole-orbitrap MS) to recognize and identify the chemical constituents, and study the quality of QG. Methods: The 50% methanol extract was separated on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm), and eluted with a gradient of methanol-water containing 0.1% formic acid. Constituents of QG were identified by HRMS in the negative ion mode using both full scan and two-stage threshold-triggered mass modes. Eleven Q-Marker were docked by Autodock vina 1.1.2 software, using H5N1 avian influenza virus as the Neuraminidase receptor, and the active site of target protein were determined by the Auto Dock 1.5.6 program. Results: A total of 39 compounds were identified, including 25 flavonoids, 6 phenylethanoid glycosides, 4 organic acids, 2 triterpenoids, and 2 lignan ingredients. Totally 15 components were specific and 32 chemical compositions were reported for the first time in QG by the method of UPLC-quadrupole-orbitrap MS. Baicalin, wogonoside, forsythoside A, liquiritin, forsythoside E, forsythoside B, isoliquiritin, baicalein, phillyrin, wogonin, and liquirtigenin could be used to establish the Q-Marker database. Conclusion: To establish Q-Marker database for Chinese medicine QG and identify the compounds in QG by the method of UPLC-quadrupole-orbitrap MS.
ABSTRACT
This study is to establish the HPLC specific chromatogram and determine four main effective components of Lamiophlomis Herba and its counterfeit.Chlorogenic acid, forsythoside B, acteoside and luteoloside were reference substance.HPLC analysis was performed on a Waters XSelect C₁₈ column (4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile-0.5% phosphoric acid solution (18∶82) with isocratic elution.The flow rate was 1.0 mL•min⁻¹, the detection wavelength was 332 nm and the column temperature was 30 ℃.Chemometrics software Chempattern was employed to analyze the research data.HPLC specific chromatogram of Lamiophlomis Herba from different samples were of high similarity, but the similarity of the HPLC specific chromatogram of its counterfeit were less than 0.65.Both of cluster and principal component analysis can distinguish certified products and adulterants.The HPLC specific chromatogram and contents of four effective components can be used for the quality control of Lamiophlomis Herba and its preparations.It provided scientific basis to standardize the use of the crude drug.
ABSTRACT
Objective: An UHPLC-MS/MS method was developed for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic parameters for three phenolic glycosides were calculated as well. Methods: Samples of plasma of rats were obtained at different time after rats were administrated with Callicarpa nudiflora extract (5 g/kg). After the addition of acidification (hydrochloric acid, 0.25 mol/L) and deproteinization by acetonitrile, plasma samples were separated on a Phenomenex® Kinetex C18 column (50 mm × 2.1 mm, 1.7 μm) with gradient elution using acetonitrile-0.005% formic acid as mobile phase. Mass spectrometric detection was carried out by multiple reaction monitoring (MRM) using electrospray ionization in negative ion mode. Results: A good linearity of acteoside, isoacteoside, and forsythoside B was shown in the ranges of 7.77 - 3 880.00 ng/mL (r2 = 0.995 5), 5.04 - 2 520.00 ng/mL (r2 = 0.994 9), and 1.78 - 890.00 ng/mL (r2 = 0.995 1), respectively. The mean extraction recoveries of analytes were in the range of 75.2% - 89.9%, and the intra- and inter-day RSD values were less than 8.8%. The tmax of acteoside, isoacteoside, and forsythoside B was about 30 min, AUC0~t were (93 881.65 ± 18 326.65), (29 204.97 ± 8 499.88), and (15 027.05 ± 3763.82) ng∙min/mL, Cmax were (2 179.00 ± 355.60), (737.57 ± 210.31), and (227.30 ± 48.38) ng/mL, t1/2z were (235.41 ± 117.90), (151.56 ± 49.23), and (161.68 ± 63.92) min, respectively. Conclusion: The method is proved to be simple, rapid, and specific, and to be suitable for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic study.
ABSTRACT
Objective To establish a method for the isolation and preparation of forsythoside B and acteoside reference substances from Lamiophlomis rotata.Methods Forsythoside B and acteoside in L.rotata were isolated and purified by macroporous resin,Sephadex column,and preparative HPLC.Results Analysis with HPLC showed the content of the prepared acteoside and forsythoside B reached to 98.93 and 99.91%,respectively.Conclusion This method is effective for the high purity of prepared acteoside and forsythoside B.It can be used as reference substances for the qualitative and quantitative analyses of Chinese herbal medicine.