Pharmacokinetic assessment of tacrolimus in combination with deoxyschizandrin in rats.

Background: Tacrolimus (Tac) is commonly used for postoperative immunosuppressive therapy in transplant patients. However, problems, for example, low bioavailability and unstable plasma concentration, persist for a long time, Studies have reported that the deoxyschizandrin could effectively improve these problems, but the pharmacokinetic parameters (PKs) of Tac combined with deoxyschizandrin are still unknown. Method: In this study, an UHPLC-MS/MS method has been established for simultaneous quantitation of Tac and deoxyschizandrin. The PKs of Tac influenced by different doses of deoxyschizandrin after single and multiple administrations were analyzed, and the different impact of deoxyschizandrin and Wuzhi capsule on PKs of Tac were compared. Result: The modified UHPLC-MS/MS method could rapid quantification of Tac and deoxyschizandrin within 2 min using bifendatatum as the internal standard (IS). All items were successfully validated. The C max of deoxyschizandrin increased from 148.27 ± 23.20 to 229.13 ± 54.77 ng/mL in rats after multiple administrations for 12 days. After co-administration of 150 mg/mL deoxyschizandrin, Tac had an earlier T max and greater C max and AUC0-t, and the C max and AUC0-t of Tac increased from 14.26 ± 4.73 to 54.48 ± 14.37 ng/mL and from 95.10 ± 32.61 to 315.23 ± 92.22 h/ng/mL, respectively; this relationship was positively proportional to the dosage of deoxyschizandrin. In addition, compared with Wuzhi capsule, the same dose of deoxyschizandrin has a better effective on Tac along with more stable overall PKs. Conclusion: An UHPLC-MS/MS method was established and validated for simultaneous detection of deoxyschizandrin and Tac. Deoxyschizandrin could improve the in vivo exposure level and stability of Tac, besides, this effect is better than Wuzhi capsule in same dose.

Deoxyschizandrin ameliorates obesity and non-alcoholic fatty liver disease: Involvement of dual Farnesyl X receptor/G protein-coupled bile acid receptor 1 activation and leptin sensitization.

Natural dual farnesyl X receptor (FXR)/G protein-coupled bile acid receptor 1 (TGR5) activators have received little attention in the management of metabolic diseases. Deoxyschizandrin (DS), a natural lignan, occurs in S. chinensis fruit and has potent hepatoprotective effects, whereas its protective roles and mechanisms against obesity and non-alcoholic fatty liver disease (NAFLD) are largely elusive. Here, we identified DS as a dual FXR/TGR5 agonist using luciferase reporter and cyclic adenosine monophosphate (cAMP) assays. DS was orally or intracerebroventricularly administrated to high-fat diet-induced obesity (DIO) mice, and methionine and choline-deficient L-amino acid diet (MCD diet)-induced non-alcoholic steatohepatitis to evaluate its protective effects. Exogenous leptin treatment was employed to investigate the sensitization effect of DS on leptin. The molecular mechanism of DS was explored by Western blot, quantitative real-time PCR analysis, and ELISA. The results showed that DS activated FXR/TGR5 signaling and effectively reduced NAFLD in DIO and MCD diet-fed mice. DS countered obesity in DIO mice by promoting anorexia and energy expenditure and reversing leptin resistance, involving both peripheral and central TGR5 activation and leptin sensitization. Our findings indicate that DS may be a novel therapeutic approach for alleviating obesity and NAFLD through regulating FXR and TGR5 activities and leptin signaling.

Deoxyschizandrin treats mice with ulcerative colitis possibly via the TLR4/NF-κB signaling pathway.

OBJECTIVE: This study aimed to investigate the effects and mechanisms of deoxyschizandrin (DSD) on treatment of ulcerative colitis (UC). METHODS: The models of mice with UC were established through dextran sulfate sodium (DSS) administration, and the successful models were treated with DSD. The therapeutic effects of DSD on UC mice were evaluated and its behind mechanisms were analyzed. RESULTS: After DSS induction, the mice showed increased body weight and colon length, worse disease activity index (DAI) and body inflammation, oxidative stress injury and increased apoptosis of colonic epithelial cells, which were remarkably relieved after DSD intervention. Besides, the levels of TLR4, MyD88 and NF-κB in the colon tissues were elevated in UC mouse models, while DSD treatment reduced the levels of these markers. CONCLUSION: DSD can alleviate the symptoms of mice with DSS-induced UC via inhibiting body inflammation, improving oxidative stress and reducing the apoptosis of colonic epithelial cells, which may be attributed to DSD inhibition of the TLR4/NF-κB signaling pathway.

BaZiBuShen alleviates altered testicular morphology and spermatogenesis and modulates Sirt6/P53 and Sirt6/NF-κB pathways in aging mice induced by D-galactose and NaNO2.

ETHNOPHARMACOLOGICAL RELEVANCE: Sperm infertility and testicular atrophy are symptoms associated with aging. BaZiBuShen formula (BZBS), a patented Chinese herbal prescription composed of Semen Cuscutae, Fructus Lycii, Epimedii Folium, Fructus Schisandrae Sphenantherae, Fructus Cnidii, Fructus Rosae Laevigatae, Semen Allii Tuberosi., Radix Morindae Officinalis, Herba Cistanches, Fructus Rubi, Radix Rehmanniae Recens, Radix Cyathulae, Radix Ginseng, Cervi Cornu Pantotrichum, Hippocampus, and Fuctus Toosendan, has been used as a kidney-tonifying and anti-aging drug as well as for the treatment of impotence and male infertility in traditional Chinese medicine. AIM OF THE STUDY: We aimed at investigating whether BZBS preserves sperm and testes morphology in aging mice, and to explore the underlying mechanisms. MATERIALS AND METHODS: BZBS was orally administered to aging mice induced by D-galactose (D-gal) and NaNO2 for 65 days. Sperm quality and testes pathophysiological alterations were examined by a Semen Analysis System, hematoxylin-eosin staining, transmission electron microscopy, and mitochondrial complex IV activity. In addition, serum levels of total antioxidant capacity (TAC), malondialdehyde (MDA), 8-hydroxy-desoxyguanosine (8-OH-dG), reduced glutathione (GSH), oxidized glutathione disulfide (GSSG), testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and tumor necrosis factor-α (TNF-α) were determined by ELISA. The expressions of P450 aromatase (CYP19), sirtuin 6 (Sirt6), P53, inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB)-p65, and phospho-NF-κB-p65 (NF-κB-pp65) in the testes were examined by western blot and/or immunohistochemical staining. RESULTS: Sustained exposure to D-gal/NaNO2 caused a deterioration of sperm quality and testes morphology in this rapid aging mouse model. BZBS treatment curtailed these alterations. These beneficial effects were associated with increased serum levels of TAC, GSH/GSSG, T, E2, and FSH, and decreased levels of MDA, TNF-α, and 8-OH-dG. BZBS treatment also downregulated the expressions of P53, iNOS, and NF-κB-pp65, as well as upregulated the expressions of Sirt6 and CYP19 in aging testes. CONCLUSIONS: BZBS preserves testicular morphology and spermatogenesis possibly via inhibition of oxidative stress and the modulation of the Sirt6/P53 and Sirt6/NF-κB signaling pathways. The results shed light on the beneficial effect of BZBS on sperm quality and fertility in aging males.

The effect of deoxyschizandrin on chronic unpredictable mild stress-induced depression.

The purpose of the present study was to evaluate the antidepressant effect of deoxyschizandrin (DEO) in chronic unpredictable mild stress (CUMS)-induced mice. The mice were subjected to CUMS paradigm for 8 weeks. From the sixth week, the mice were intragastrically treated with DEO once daily for continuous 3 weeks. The behavior tests including sucrose preference test (SPT), forced swimming test (FST), tail suspension test (TST), and open field test were conducted. Additionally, the expressions of TLR4, MyD88, TRAF6, p-NF-κBp65, NLRP3, cleaved caspase-1, cleaved IL-1ß, GluR, and PSD95 in hippocampus were detected by western blot. The concentrations of IL-6 and TNF-α in hippocampus were determined by enzyme linked immune sorbent assay (ELISA). The dendritic spine density was observed by Golgi-Cox staining. As a result, the treatment with DEO relieved anhedonia in SPT, and reduced immobile duration in FST and TST. DEO treatment effectively attenuated the CUMS-caused alterations of TLR4, MyD88, TRAF6, p-NF-κBp65, NLRP3, cleaved caspase-1, cleaved IL-1ß, GluR, and PSD95. Furthermore, DEO could reduce the hippocampal inflammatory cytokine content and increase the density of dendritic spine. In conclusion, the present work indicated that DEO exhibited antidepressant effect on CUMS-induced depressive mice, which was possible due to the TLR4/NF-κB/NLRP3 pathway and the amelioration of dendritic spine density through GluR/PSD95 cascade.

Establishment of HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneous determination of six constituents / 国际药学研究杂志

Objective: To establish HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneously determine the contents of the six constituents, acteoside, martynoside, calycosin 7-O-β- D-glucopyranoside, schisandrin, formononetin and deoxyschizan- drin. Methods: The analysis of 50% methanol extract of this drug was performed on a 30℃ thermostatic Waters Symmetry C18 column, with the mobile phase comprising of acetonitrile(A)-0.2% formic acid solution(B)flowing at 1.0 ml/min in a gradient elution manner. The detection wavelength was set at 330 nm for acteoside and martynoside, 254 nm for calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin. Results: There were thirteen common peaks in the fingerprints of ten batches of samples with the similarities more than 0.9. Acteoside, martynoside, calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin showed good linear relationship within the ranges of 1.47-36.75, 0.66-16.50, 0.89-22.25, 2.58-64.50, 1.91-47.75 and 0.77-19.25 μg/ml(r≥0.9993), and their average recoveries were 97.57%, 99.22%, 96.69%, 100.01%, 98.79% and 96.77%, with the RSD of 0.79%, 1.54%, 0.61%, 0.64%, 0.83% and 0.36%, respectively. Conclusion: The established method is easily operable and repeatable, which could be used for quality control of Qiwei Tangmaishu tablet.

Synergy effect of deoxyschizandrin and gemcitabine on proliferation of HepG2 human hepatocellular carcinoma cells / 中国药理学通报

Aim To investigate the synergy effects of deoxyschizandrin and gemcitabine (GEM) on the proliferation of HepG2 human hepatocellular carcinoma cells in vitro and the underlying mechanism. Methods CCK8 method and colony formation assay were used to detect the effects of deoxyschizandrin monotherapy, GEM monotherapy and combination therapy on the proliferation of HepG2 cells. Flow cytometry was used to detect the change of apoptosis rate of HepG2 cells after treatment with single drug or the combination use of two drugs. Western blot was performed to detect the expression of BCL2, BAX, pro-caspase3, caspase3, pro-caspase9, caspase9, ß-catenin and TCF-4. Results Deoxyschizandrin, GEM and combination group significantly inhibited the proliferation of HepG2 cells and promoted cell apoptosis. The effects of the combination group on HepG2 cells were significantly stronger than those of single-drug groups (P < 0. 05). Western blot results showed the expression of pro-caspase3 and pro-caspase9 was changed slightly within deoxyschizandrin, GEM and combination groups compared with that of normal control, while the expression of B C L 2, ß-catenin and TCF-4 protein expression was down-regulated significantly (P < 0. 05). The expression of B A X, cleaved-caspase3 and cleaved-caspase 9 protein increased significantly after treatment with deoxyschizandrin, GEM and combination group (P < 0. 05). Specially, the increasing effect of the expression of the protein in combined group was more significant than that of single drug groups (P < 0. 05). Conclusions The combination of deoxyschizandrin and GEM significantly inhibited the proliferation of HepG2 cells and induced cell apoptosis, as well as suppressed the ß-catenin/TCF-4 pathway.

Study on the Effects of Deoxyschizandrin on the Proliferation ，Migration and Invasion of Human Nasopharyngeal Carcinoma Cell HONE- 1 via Met/PI 3K/Akt Signaling Pathway / 中国药房

OBJECTIVE：To study the effects and potential mechani sm of deoxyschizandrin on the proliferation ，migration and invasion of nasopharyngeal carcinoma cell HONE- 1. METHODS ：HONE-1 cell was set as cell model ，while CCK- 8 test，wound healing assay and Transwell chamber test were used to detect the proliferation ，migration and invasion ability changes of HONE- 1 cells after treatment with different concentrations [ 0（blank control ），10，20，40 μmol/L] of deoxyschizandrin. Computer molecular docking was performed to analyze the binding ability between deoxyschizandrin and Met protein. Western blotting assay was used to detect the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells. RESULTS ：Compared with blank control ，the proliferation ，migration and invasion ability of cells after treated with 10，20，40 μmol/L deoxyschizandrin were all decreased significantly （P＜0.05）. Results of molecular docking revealed that deoxyschizandrin could stably bind with the activity pocket of Met protein. Results of Western blotting assay demonstrated that compared with blank control ，10，20，40 μmol/L deoxyschizandrin all decreased the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells significantly（P＜0.05）. CONCLUSIONS ：Deoxyschizandrin can inhibit the proliferation ，migration and invasion of HONE- 1 cell via inhibiting the activation of Met/PI 3K/Akt signaling pathway.

Interaction of deoxyschizandrin and schizandrin B with liver uptake transporters OATP1B1 and OATP1B3.

1. Deoxyschizandrin and schizandrin B have diverse pharmacological effects, including hepatoprotective activity. We aim to study their hepatic uptake and their effects on the hepatic uptake of other clinical drugs mediated by OATP1B1 and OATP1B3. 2. Deoxyschizandrin exhibited a high affinity for OATP1B1 with Km of 17.61 ± 0.43 µM but a low affinity for OATP1B3. Similarly, schizandrin B also showed a strong affinity for OATP1B1 with Km of 18.45 ± 1.23 µM but a weak affinity for OATP1B3. 3. Atorvastatin and rifampicin could inhibit the uptake of deoxyschizandrin and schizandrin B mediated by OATP1B1. 4. Intriguingly, both deoxyschizandrin and schizandrin B significantly promoted the uptake of atorvastatin (with EC50 of 50.58 ± 8.08 and 24.70 ± 5.82 µM, respectively) and rosuvastatin (with EC50 of 13.46 ± 2.70 and 8.99 ± 4.73 µM, respectively) mediated by OATP1B1. Deoxyschizandrin could markedly promote the uptake of fluvastatin but inhibit the uptake of sodium taurocholate (TCNa) mediated by OATP1B1. 5. The promotion on hepatic uptake of statins mediated by OATP1B1 might lead to enhanced efficacy of cholesterol lowering and reduced risk of myopathy for hyperlipidemia patients when given statins together with deoxyschizandrin or schizandrin B.

Prediction and analysis of Q-markers between crude and vinegar Schisandrae Chinensis Fructus based on multivariate statistical analysis and network pharmacology / 中草药

Objective: To explore the potential Q-markers between crude Schisandrae Chinensis Fructus (SCF) and vinegar-processed Schisandrae Chinensis Fructus (VSCF) based on multivariate statistical analysis and network pharmacology. Methods: UPLC-Q/TOF-MS was used to analyze the main lignans in SCF and VSCF, and the potential differences of chemical components (Q-markers) between SCF and VSCF were screened out by using multiple statistical methods. Furthermore, through network pharmacology and bioinformatics, the main action targets and pathways related to significantly different components were analyzed to construct the “component-target-pathway” network relationship and predict the potential quality markers between SCF and VSCF. Results: In this study, 40 different constituents of Schisandra chinensis between SCF and VSCF were screened, among which eight chemical markers had significant differences between SCF and VSCF. Five chemical constituents were identified and confirmed, namely 5-HMF, deoxyschizandrin and its isomer, schisandrin B, and schisantherin D. The other three chemical markers were speculated to be lignans by analyzing the first-and second-order mass spectrometry information. The results of network pharmacological analysis showed that the five potential quality markers identified were highly related to the main pharmacological effects of SCF. Finally, schisandrin B and 5-hydroxymethyl furfural were identified as the most representative potential quality markers. Conclusion: The results showed that the chemical composition of SCF had a series of complex changes. It was determined that schisandrin B and 5-hydroxymethyl furfural could be used as representative potential quality markers between SCF and VSCF. It is speculated that lignans may be the basis of the important effect of VSCF on liver protection.

Deoxyschizandrin Loaded Liposomes on the Suppression Lipid Accumulation in 3T3-L1 Adipocytes.

Deoxyschizandrin (DS) is a bioactive benzocyclooctadiene lignan found in the fruit of Schisandra chinensis. However, poor bioavailability and non-specificity of DS frequently caused low therapeutic efficacy. In the present study, DS-liposome (DS-lipo) was implemented to enhance the hepatic targeting and inhibition effects on adipocyte differentiation in 3T3-L1 cells. The formulations enabled encapsulation of as much as 24.14% DS. The DS-lipo prepared was about 73.08 nm, as measured by laser light scattering (LLS) morphology. In the visual field of a scanning electron microscope (SEM), the liposomes were spherical with similar size and uniform dispersion. Fluorescence live imaging study exhibited hepatic targeting of liposomes in vivo. Furthermore, High-Content Analysis (HCS) imaging microassay analyses revealed DS-lipo and DS reduced cytoplasmic lipid droplet in 3T3-L1 adipocytes, with the IC50 value of 8.68 µM and 31.08 µM, respectively. The lipid droplet accumulation inhibition rate of 10 µM DS-lipo was above 90%, which was even superior to the effect of 30 µM DS solution. The current findings suggest that DS-lipo was a therapeutic strategy for alleviating lipid-associated diseases and nonalcoholic fatty liver disease (NAFLD).

Deoxyschizandrin, Isolated from Schisandra Berries, Induces Cell Cycle Arrest in Ovarian Cancer Cells and Inhibits the Protumoural Activation of Tumour-Associated Macrophages.

Deoxyschizandrin, a major lignan of Schisandra berries, has been demonstrated to have various biological activities such as antioxidant, hepatoprotective, and antidiabetic effects. However, the anti-cancer effects of deoxyschizandrin are poorly characterized. In the present study, we investigated the anti-cancer effect of deoxyschizandrin on human ovarian cancer cell lines and tumour-associated macrophages (TAMs). Deoxyschizandrin induced G0/G1 phase cell cycle arrest and inhibited cyclin E expression in human ovarian cancer cells. Overexpression of cyclin E significantly reversed the deoxyschizandrin-induced cell growth inhibition. Interestingly, increased production of reactive oxygen species and decreased activation of Akt were observed in A2780 cells treated with deoxyschizandrin, and the antioxidant compromised the deoxyschizandrin-induced cell growth inhibition and Akt inactivation. Moreover, deoxyschizandrin-induced cell growth inhibition was markedly suppressed by Akt overexpression. In addition, deoxyschizandrin was found to inhibit the expression of the M2 phenotype markers CD163 and CD209 in TAMs, macrophages stimulated by the ovarian cancer cells. Moreover, expression and production of the tumour-promoting factors MMP-9, RANTES, and VEGF, which are highly enhanced in TAMs, was significantly suppressed by deoxyschizandrin treatment. Taken together, these data suggest that deoxyschizandrin exerts anti-cancer effects by inducing G0/G1 cell cycle arrest in ovarian cancer cells and reducing the protumoural phenotype of TAMs.

Study on the pharmacokinetics of deoxyschizandrin and schizandrin in combination with epigallocatechin gallate, a component of green tea, in rats.

1. Green tea is commonly used worldwide due to its potential positive health benefits. We have examined the effects of epigallocatechin gallate (EGCG), the most abundant catechin in green tea, on the pharmacokinetics of deoxyschizandrin (DSD) and schizandrin (SD), which are the representative lignans in popular traditional Chinese medicines Fructus schisandrae, in rats. 2. The effects on the transport in Caco-2 cells and metabolism in human liver microsomes (HLMs) of DSD and SD by EGCG were determined to analyze their interactions thoroughly. 3. In pharmacokinetic studies, rats were divided into four groups. Each group was orally treated with DSD alone (Group 1), DSD combined with EGCG (Group 2), SD alone (Group 3) and SD combined with EGCG (Group 4). The pharmacokinetic parameters of DSD and SD in rats were determined by UPLC-MS/MS. 4. The in vivo results indicated that EGCG had no significant influence on the pharmacokinetic behaviors of DSD or SD in rats, which were in accordance with the in vitro transport and metabolism studies. However, there were marked differences between male and female rats among Cmax, AUC0-t, AUC0-∞ of DSD and SD. This disparity suggested that gender differences might exist in the pharmacokinetic processes of DSD or SD in rats.

Simultaneous determination of nine bioactive components in Sishen Pills by HPLC-ESI-MS/MS / 中草药

Objective To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components (deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine) in Sishen Pills. Methods The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C18 column (100 mm × 4.6mm, 3.5 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.4 mL/min, and the injection volume was 20 μL. The nine major bioactive components were detected using an electrospray ionization source in positive ionization mode (ESI+) and quantified by multiple reaction monitor (MRM) scanning at the same time. Results The linear ranges of deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine were 8.50-850.00 ng/mL (r = 0.999 7), 1.32-132.00 ng/mL (r = 0.997 4), 9.60-960.00 ng/mL (r = 0.999 8), 12.00-1 200.00 ng/mL (r = 0.999 3), 11.50-1 150.00 ng/mL (r = 0.997 9), 21.70-2 170.00 ng/mL (r = 0.999 7), 23.80-2 380.00 ng/mL (r = 0.999 6), 10.70-1 070.00 ng/mL (r = 0.999 5), 8.54-854.00 ng/mL (r = 0.998 0), and the average recoveries were 98.3% (RSD = 2.21%), 100.3% (RSD = 1.78%), 99.2% (RSD = 2.19%), 100.4% (RSD = 2.23%), 99.1% (RSD = 2.18%), 97.7% (RSD = 3.03%), 99.0% (RSD = 2.51%), 98.9% (RSD = 2.72%), and 100.3% (RSD = 2.10%), respectively. The contents of eight batches of the nine major bioactive components were 67.6-425.6, 0-131.5, 2.1-258.0, 0-71.2, 23.2-678.8, 806.4-1310.8, 718.5-1293.7, 11.5-123.2, and 10.9-62.4 μg/g, respectively. Conclusion The developed method is simple, specific, and sensitive, and it can be applied for the determination of nine major bioactive components and the quality control of Sishen Pills collected from different production batches.

Quality evaluation of Liver-protection Agent based on HPLC fingerprint and simultaneous determinations of multiple components / 中草药

Objective To establish HPLC fingerprint strategy to simultaneously determine nine components in Liver-protection Agent, including loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid, and provide a scientific basis for the quality control of Liver-protection Agent and related medicinal preparations. Methods The analytical analysis was performed on an Agilent 1260-HPLC system equipped with a VWD detector and SB-C18 reversed phase column (Zorbax 150 mm × 4.6 mm, 5 μm). The analytes were eluted using a gradient mixture of two solvent: solvent A was distilled deionized water containing 0.1% ortho-phosphoric acid and solvent B was 100% acetonitrile. The mobile phase flow rate was 1.0 mL/min. The separation temperature was 30 ℃. The detection wavelengths were 236, 280, and 210 nm. The injection volume was 5 μL. Common patterns of HPLC fingerprints for 10 batches of Liver-protection Agent medicinal preparations were established, and chemometric analysis method was employed to analyze the hidden information. At the same time, methodological study was conducted for determinations of multiple components including loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid. Results The HPLC fingerprint strategy of Liver-protection Agent medicinal preparation had been set up, and 37 common peaks had been identified with the similarity of more than 0.9. Moreover, the samples were roughly divided into four categories by the methods of the systematic cluster analysis and principle component analysis. After validating the multiple component quantitative analysis condition through methodology, the average recovery rate was between 95.13% and 104.8%.The average mass concentration of loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid in ten batches of Liver-protection Agent was 216.3-223.0, 126.1-137.1, 144.7-149.0,1 623.1-1 992.7, 481.9-520.0, 14.9-18.7, 33.8-37.0, 2.9-3.7, and 39.7-43.6 mg/L, respectively. Conclusion The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are rapid, simple, and reproducible, which can be adopted as one of the quality control methods for Liver-protection Agent and related medicinal preparations.

Simultaneous determination of six constituents in Maiwei Dihuang Pills by HPLC-DAD / 中成药

AIM To establish an HPLC-DAD method for the simultaneous content determination of six constituents in Maiwei Dihuang Pills (Ophiopogonis Radix,Schisandrae chinensis Fructus,Rehmanniae Radix Praeparata,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent ZORBAX SB-C18column (4.6 mm × 150 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.2％ phosphate acid) flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,230,236 and 274 nm.RESULTS Deoxyschizandrin,schizandrin B,schisandrin,paeoniflorin,paeonol and loganin showed good linear relationships within the ranges of 10-70,6.5-45.5,33.5-234.5,17-119,31-217 and 34-238 μg/mL (r ＞0.990 0),whose average recoveries (RSDs) were 99.6％ (1.7％),100.4％ (1.8％),100.7％ (1.8％),102.9％ (1.7％),102.2％ (1.5％) and 99.7％ (1.2％),respectively.CONCLUSION This simple and reproducible method can be used for the rapid quality control of Maiwei Dihuang Pills.

HPLC fingerprint analysis on Qibai Pingfei Granules and multi-components determination / 中草药

Objective: To establish an HPLC fingerprint of the compounds in Qibai Pingfei Granules (QPG), and to make a quantitative analysis. Methods: Sample was extracted by 50% methanol. Phenomenex Luna C18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of methanol-0.2% formic acid gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 250 nm, and the column temperature was 30℃. The chemical component fingerprint similarity of 10 batches of QPG was calculated with Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012) published by National Pharmacopoeia Committee and the common peaks were identified by reference compounds. Results: Fingerprints of 10 batches of QPG were established and the similarities to the common mode were above 0.96. Totally 25 common peaks were found. Among them, peak 1 belonged to Pheretima, peaks 2 and 3 belonged to Pheretima and Ginseng Radix et Rhizoma (GRR), peak 4 belonged to GRR, Schisandrae Chinensis Fructus (SCF), and Chuanxiong Rhizoma (CR), peak 5 belonged to GRR, Allii Macrostemonis Bulbus (AMB), CR, and Pheretima, peaks 6, 8, 22, 23, 24, and 25 belonged to SCF, peak 7 belonged to Astragali Radix (AR) and SCF, peak 9 belonged to GRR and AMB, peak 10 belonged to Descurainiae Semen Lepidii Semen (DSLS) and AMB, peaks 11, 12, 13, 15, and 16 belonged to CR, peak 14 belonged to AR and DSLS, peaks 17, 18, 19, 20, and 21 belonged to AR. Based on the retention time, and UV absorption spectra of reference compounds, six constituents including caffeic acid (peak 12), ferulic acid (peak 13), schizandrol A (peak 22), schisantherin A (peak 23), deoxyschizandrin (peak 24), and schisandrin (peak 25) were identified. The linear ranges of caffeic acid, ferulic acid, schizandrol A, schisantherin A, deoxyschizandrin, and schisandrin were 3.38-108.02, 3.60-115.33, 2.99- 95.61, 2.81-89.77, 3.26-104.17, and 2.89-92.45 μg/mL, respectively. In 10 batches of QPG samples, the contents were as follows: caffeic acid of 0.412-0.429 mg/g, ferulic acid of 0.302-0.317 mg/g, schizandrol of A 0.182-0.195 mg/g, schisantherin A of 0.179-0.195 mg/g, deoxyschizandrin of 0.203-0.215 mg/g, and schisandrin of 0.131-0.144 mg/g, the amount of each indicator composition among different batches changed a litte, and the sample quality is stable. Conclusion: The method has good precision, reproducibile, stability, separation, and can be used for the quality control of QPG.

Correlation between color and chemical constituents of fruits of Schisandra chinensis / 中草药

Objective: To study the relativity between color and chemical composition in fruits of Schisandra chinensis by fruits' surface color value (L*, a*, and b*) and contents of active ingredients of fructus core S. chinensis). Methods: The surface chromatic value (multivariate linear regression equation of L*, a*, b* values and deoxyschizandrin, schizandrin B, schizandrol A, schinsantherin A) was established by SPSS 20.0 with mathematical statistics method, then the influence degree of the active ingredients of. S. chinensis was detected by F test. Results: The value of L* were significant negative with deoxyschizandrin (r ≥ 0.5), and other chromaticity values were very low or low-grade correlated with lignans Conclusion: The effective ingredient content of fruit of S. chinensis can be estimated by its chromatic values, and also can be used in the hierarchies of the fruit of S. chinensis, in order to provide better scientific basis of the quality of S. chinensis.

Application of QAMS in quality control of Wuzi Yanzong Pills / 中草药

Objective To establish a quantitative analysis of multi-component with a single-marker (QAMS) method for the quality control of Wuzi Yanzong Pills (WYP). Methods Six main effective components (schisandrin, hyperin, quercitrin, kaempferol 3-O-rutinoside, deoxyschizandrin, and γ-schizandrin) of WYP were simultaneously separated on a reversed-phase column (Ultimate LP-C18) with high-resolution of each chromatographic peak by high performance liquid chromatography (HPLC). Schisandrin was selected as the internal reference, and the relative correlation factors (RCFs) of other five components were calculated to achieve QAMS. The ruggedness of RCFs was tested on different instruments and columns. Moreover, results of the QAMS were compared with the external standard method. Results Within a certain linear range, the RCFs of hyperin, quercitrin, kaempferol 3-rutinoside, deoxyschizandrin, and γ-schizandrin were 0.36, 4.86, 0.88, 7.34, and 6.35, respectively. The repeatability was good under different experimental conditions. There were no significant differences between the calculated value and estimated value on QAMS and external standard method. Conclusion The QAMS method can be used to assay the content of six components of WYP simultaneously and control the quality of WYP simplely, reliably, and accurately.

nhibitory effect and mechanism of deoxyschizandrin on NLRP3 inflammasome / 药学学报

This study was conducted to investigate the inhibitory effect and the molecular mechanism of deoxyschizandrin on the activity of NLRP3 (NOD-like receptor family, pyrin domain containing 3) inflammasome. Bone marrow-derived macrophages were used to study the effects of deoxyschizandrin on inflammasome activation using inflammasome inducers (ATP and nigericin). Cytotoxic effect was evaluated with CCK-8. The expression of IL-1β, caspase-1 in the supernatant and the expression of pro-caspase-1, pro-IL-1 β, ASC, NLRP3 in cell was detected by Western blot for the inhibitory effect of deoxyschizandrin (25, 50, 100 and 200 μmol·L-1) on the activity of NLRP3 inflammasome. Immunofluorescence was applied to investigate NF-κB (p65) transportation to the nucleus. The results of CCK-8 showed that the optimum concentration of deoxyschizandrin was 6.25-400 μmol·L-1. Deoxyschizandrin (25, 50, 100, and 200 μmol·L-1) could inhibit the activation of NLRP3 inflammasome caused by nigericin and ATP, and inhibit the secretion of IL-1 β, which was associated with inhibiting the cleavage of pro-caspase-1. The results of immunofluorescence and Western blot also suggest that the inhibitory activity of deoxyschizandrin on NLRP3 inflammasome was not dependent on NF-κB pathway and protein expression of NLRP3, ASC, pro-caspase-1 and pro-IL-1 β mediated by NF-κB. Our results confirmed that deoxyschizandrin could suppress the cleavage of pro-caspase-1 and inhibit the activity of NLRP3 inflammasome at 25-200 μmol·L-1 to reduce the inflammation response.