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OBJECTIVE@#To investigate the anti-neuroinflammation effect of extract of Fructus Schisandrae chinensis (EFSC) on lipopolysaccharide (LPS)-induced BV-2 cells and the possible involved mechanisms.@*METHODS@#Primary cortical neurons were isolated from embryonic (E17-18) cortices of Institute of Cancer Research (ICR) mouse fetuses. Primary microglia and astroglia were isolated from the frontal cortices of newborn ICR mouse. Different cells were cultured in specific culture medium. Cells were divided into 5 groups: control group, LPS group (treated with 1 μg/mL LPS only) and EFSC groups (treated with 1 μg/mL LPS and 100, 200 or 400 mg/mL EFSC, respectively). The effect of EFSC on cells viability was tested by methylthiazolyldiphenyltetrazolium bromide (MTT) colorimetric assay. EFSC-mediated inhibition of LPS-induced production of pro-inflammatory mediators, such as nitrite oxide (NO) and interleukin-6 (IL-6) were quantified and neuron-protection effect against microglia-mediated inflammation injury was tested by hoechst 33258 apoptosis assay and crystal violet staining assay. The expression of pro-inflammatory marker proteins was evaluated by Western blot analysis or immunofluorescence.@*RESULTS@#EFSC (200 and 400 mg/mL) reduced NO, IL-6, inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) expression in LPS-induced BV-2 cells (P<0.01 or P<0.05). EFSC (200 and 400 mg/mL) reduced the expression of NO in LPS-induced primary microglia and astroglia (P<0.01). In addition, EFSC alleviated cell apoptosis and inflammation injury in neurons exposed to microglia-conditioned medium (P<0.01). The mechanistic studies indicated EFSC could suppress nuclear factor (NF)-?B phosphorylation and its nuclear translocation (P<0.01). The anti-inflammatory effect of EFSC occurred through suppressed activation of mitogen-activated protein kinase (MAPK) pathway (P<0.01 or P<0.05).@*CONCLUSION@#EFSC acted as an anti-inflammatory agent in LPS-induced glia cells. These effects might be realized through blocking of NF-κB activity and inhibition of MAPK signaling pathways.
Subject(s)
Animals , Astrocytes , Metabolism , Pathology , Cell Line , Cell Nucleus , Metabolism , Chromatography, High Pressure Liquid , Down-Regulation , Inflammation , Pathology , Inflammation Mediators , Metabolism , Lipopolysaccharides , MAP Kinase Signaling System , Mice, Inbred ICR , Microglia , Metabolism , Pathology , NF-kappa B , Metabolism , Nervous System , Pathology , Neurons , Metabolism , Pathology , Neuroprotective Agents , Pharmacology , Plant Extracts , Pharmacology , Schisandra , Chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
This study was performed to use UHPLC-QTOF/MSE technology to rapidly search and identify variations of chemical ingredients between Fructus Schisandrae Chinensis and its processed products. The present study provides a basis for the study of Chinese herbal medicine processing with a focus on the impact of processing on chemical components. Using a time-dependent data scan mode (MSE) couple with metabolomics technology, we acquired accurate data and identified the potential chemical markers. A total of 12 chemical markers were identified in the crude, vinegar-processed and wine-processed Schisandra chinensis fruit; The results showed that the levels of 6-O-benzoylgomisin O, schisantherin B, schisantherin C, schisantherin D and neokadsuranic acid are the highest in crude Schisandra chinensis fruit; thelevels of schizandrin A, schizandrin B, schizandrin C, gomisin D and gomisin T are the highest in wine-processed Schisandra chinensis fruit; the levels ofschisantherin A and schisandrin are the highest in vinegar-processed Schisandra chinensis fruit. There were significant changes of chemical components between Fructus Schisandrae Chinensis and their processed products, and these findings may offer a reasonable explanation for variation of efficacy and clinical applications in the processed products of Fructus Schisandrae Chinensis.
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Objective To prepare Wurenchun solid dispersion of alcohol extracts of Fructus Schisandrae Chinensis so as to improve the dissolution of its active compound in vitro.Methods The Wurenchun solid dispersions were prepared with various carriers and drug/carrier ratios by mixing the carrier in alcohol extractive solution of FSC directly,and the apparent solubility and dissolution of deoxyschisandrin in them were tested and compared.Results The apparent solubility and dissolution of deoxyschisandrin of Wurenchun solid dispersion(extracts: polyvinylpyrrolidone(PVP) K30=1∶3) were increased remarkably to 5.06 ?g/mL and 43.2% in water individually,including dispersed and dissolved drug whose particle size is below 0.22 ?m,compared with that of the self-prepared Wurenchun capsules.Conclusion Wurenchun solid dispersion made of PVP K30 can remarkably enhance apparent solubility and dissolution of the active compound in vitro.
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Objective To establish a HPLC method for the determination of deoxys chizandrin and schisantherrin A in Fructus Schisandrae Chinensis. Methods The as say was conducted on a YWG-C18 column with methonal-water (77 ∶23) as mobile phase. The flow rate was 1.0 mL/min and detection wavelength was 230 nm. Results Satisfactory linearities of deoxyschizandrin and schisantherrin A were shown in the range of 0.125～1.25 ?g and 0.08～0.8 ?g,and the recovery was 100.1 %an d 100.0 %(RSD being 0.15 %and 0.28 %。Schisandra sphenanthcra),and 99.7 %an d 101.2 %(RSD being 0.32 %and 0.67 %。Schisandra chinensis) (n=5). Conclusion The HPLC method was simple,curate,highly sensitive and reproducible. It may b e used for the quantitative determination of deoxyschizandrin and schisantherrin A in Fructus Schisandrae Chinensis.
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Objective To establish a HPLC determination method for the effective constituents extracted from Fructus Schisandrae Chinensis by low-temperature water extraction.Methods The assay was conducted on Kromasil C18 column(250 mm? 4.6 mm,5 ? m)using a gradient elution with acetonitrile-water as mobile phase.The flow rate was set at 1.0 mL/min,column temperature was kept at 30 ℃,and the detection wavelength was at 240 nm.Results The linear range for schizandrol A was 5.0～ 200.0 ? g/mL(r=0.999 9),and the average recovery was 101.27 % with RSD being 1.97 %(n=6).Conclusion The method is simple,reliable and reproducible for the determination of schizandrol A.It can be used for quality control of preparations including Fructus Schisandrae Chinensis.
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AIM: To establish a method of determining the fingerprints of water-soluble composition in Fructus schisandrae chinensis by HPLC. METHODS: Separation was performed on Discovery SB-C18(150 mm?4.6 mm,5 ?m) analytical column with mobile phase consisting of water and acetonitrile with gradient elute at the flow rate of 1.0 mL/min.The UV detection wavelength was set at 203 nm. RESULTS: Compared with reference peak of schisandrin,8 common peaks on frngerprints of Fructus schisandrae chinensis were indicated by the method with satisfied stability,precision and repeatability. CONCLUSION: There are good similarities among 10 batches of Fructus schisandrae chinensis,and this method is reliable,simple and provides a reference standard for the quality control of Fructus schisandrae chinensis.
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AIM: To optimize the preparation process of Fructus Schisandrae Chinensis dispensing granules METHODS:Schizandrin,schisantherin A,deoxyschizandrin,r-schizandrin were viewed as the indexes of judgement criteria,the influencing factors of extraction process as extract solvent,medical material particle diameter were studied and extraction parameters were optimized. RESULTS: Active components of dispensig granules can be extracted effectively by alcohol when medical materials were crushed.The best conditions were as follows:medical material were ground into 40 mesh,extracted two times by 70% alcohol,2 h per time. CONCLUSION: After being optimized,The process of dispensing granules is better than that of products on market.
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AIM: To establish the quality standard for Huangqi Fufang Granules(Radix Astragali, Fructus Schisandrae Chinensis, Radix Cyathulae, etc.). METHODS: Fructus Schisandrae Chinensis, Radix Cyathulae were identified by TLC, and the content of astragaloside Ⅳ was determined by TLC scanning. RESULTS: Fructus Schisandrae Chinensis, Radix Cyathulae could be identified by TLC. Astragaloside Ⅳ showed a good linear relationship at a range of 1.0?g～6.0?g, r =0.9992. The average recovery was 99.19%, and RSD was 0.63%. CONCLUSION: The methods are simple, accurate and specific, and can be used for the quality control of Huangqi Fufang Granules.
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OBJECTIVE: To optimize the extraction process of Fructus Schisandrae Chinensis. METHODS: Taking the contents of schizandrin, schisantherin A, deoxyschizandrin, r-schizandrin as the indexes, the influence on the extraction process by extract solvent and medicinal material particle diameter was examined, and the extraction parameters were optimized. RESULTS: Active components of Fructus Schisandrae Chinensis, after the medicinal material being crushed, can be effectively utilized through extracting by alcohol. The optimized extraction parameters were as follows: the medical material being ground into 40 meshes, extracted twice by 70% alcohol, 2 hours per time. CONCLUSION: The optimized extracting process works better than current technique adopted for formulated granule preparation, and can ensure quality to the highest degree.