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[Identification of Fructus Schisandrae chinensis and Fructus Schisandrae sphenantherae in wu zi yan zong pill by RP-HPLC analysis].

OBJECTIVE: To identify Fructus schisandrae chinensis and Fructus schisandrae sphenantherae in wu zi yan zong pill (WZYZP). METHODS: Columm SUPELCOSIL C18; Mobile phase: methanol-water (65:35); Detection wavelength: 254 nm. RESULTS: Using above optimum chromatography conditions, the HPLC chromatography of relative medicinal materials and WZYZP was established. CONCLUSION: HPLC chromatography can be used for identification of Fructus schisandrae chinensis and Fructus schisandrae sphenantherae in WZYZP.

Chromatography, High Pressure Liquid↗

[Ribosomal DNA ITS sequences analysis of the Chinese crude drug fructus schisandrae sphenantherae and fruts of Schisandra viridis].

OBJECTIVE: To find the patterns of the rDNA ITS sequence variation of Schisandra sphenanthera and S. viridis, and to establish the molecular biological method for the identification of Fructus Schisandrae Sphenantherae and the fruits of S. viridis. METHOD: PCR products were sequenced directly and the sequences were analyzed with PAUP 4.0b10. NJ systematic tree was obtained with neighbor-joining method. RESULT: The Complete ITS sequence of S. sphenanthera was 691-692 bp, of which there were 282 bp of ITS1 and 246-247 bp of ITS2. The complete sequence of S. viridis was 694-695 bp, consisting of 285-286 bp of ITS1 and 246-247 bp of ITS2. There were three informative sites in ITS1 regions for the two species. In the NJ tree with Kadsura anamosma and K. coccinea as outgroups, five different populations of S. viridis were the monophyletic group with the bootstrap value of 68%. These populations included one from Tianmushan, Zhejiang province, three populations from Jigongshan, Henan Province and the other two populations of S. viridis cited the sequences from GeneBank (registration numbers are AF263438 and AF163703 respectively). CONCLUSION: The rDNA internal transcribed spacer is a good marker to distinguish the Fructus Schisandrae Sphenantherae from the fruits of S. viridis.

Base Sequence↗

Evaluation of the protective effects of Schisandra chinensis on Phase I drug metabolism using a CCl4 intoxication model.

To evaluate the potential activity of Schisandra chinensis in restoring hepatic drug metabolism in CCl4 damaged liver, antipyrine was employed as a probe for the possible effects of the herb on Phase I oxidative metabolism in rats. Schisandra lignan fraction (160 mg/kg) was given orally to male Sprague-Dawley rats (220-240 g) 30 min or 6 h before CCl4 intoxication (4 ml/kg, s.c.). Following a single oral dose of antipyrine (80 mg/kg) to the rats with damaged liver, the pharmacokinetics of antipyrine in whole blood were determined and levels of liver enzymes, e.g. SGPT, SGOT, and cytochrome P450 were measured. Pharmacokinetic parameters for antipyrine were estimated using noncompartmental analysis. Results indicated that CCl4 significantly increased the elimination half-life (t(1/2)) of antipyrine from 2.59 +/- 1.04 to 11.25 +/- 3.91 h (P < 0.001) and decreased its clearance (CL) from 65.94 to 10.84 ml/h as compared to control. Pretreatment with the Schisandra lignan fraction 30 min or 6 h before intoxication significantly (P < 0.001) improved antipyrine elimination by reducing its t(1/2) to 3.30 +/- 0.52 and 3.58 +/- 1.05 h, respectively. The corresponding improvements observed for CL, i.e. 49.06 +/- 21.75 ml/h (P < 0.01); 21.10 +/- 10.42 ml/h (P < 0.05), were also substantial. Moreover, normalization of SGPT, SGOT and P450 levels was observed with the two Schisandra pretreatment schedules. In conclusion, Schisandra lignans exhibited strong protective effect on Phase I oxidative metabolism in the liver damaged by CCl4. Furthermore, pretreatment of Schisandra 30 min before intoxication showed a more pronounced effect than that of the 6 h pretreatment. The current pharmacokinetic approach allowed the protective effects of Schisandra on oxidative drug metabolism in damaged liver to be systemically examined and will certainly help in the evaluation of hepato-protectants obtained from natural sources.

Animals↗

Biochemical basis of the "Qi-invigorating" action of Schisandra berry (wu-wei-zi) in Chinese medicine.

Schisandra berry or Wu-Wei-Zi, meaning the "the fruit of five tastes" in Chinese, is a commonly used herb in traditional Chinese medicine (TCM). Based on the "Five-Element" theory in TCM, while the "five tastes" of Schisandra berry refer to its influence on the five visceral organs in the body, ancient Chinese herbalists specifically trumpeted the berry's beneficial effect on the "Qi" of the five visceral organs. "Qi" is a Chinese term used as a broad description of energy-dependent body functions. Over the past ten years, our laboratory has attempted to define the biochemical properties of Schisandra berry in regard to its purported "Qi-invigorating" properties. We have found, for the first time, an ability of Schisandra berry to fortify mitochondrial antioxidant status, thereby offering the body a generalized protection against noxious challenges both of internal and external origin. Given the indispensable role of the mitochondrion in generating cellular energy, the linking of Schisandra berry to the safeguarding of mitochondrial function provides a biochemical explanation for its "Qi-invigorating" action.

Antioxidants↗

Improvement of phase I drug metabolism with Schisandra chinensis against CCl4 hepatotoxicity in a rat model.

The seed extract of Schisandra chinensis was investigated in the rat for its restorative or therapeutic effect on Phase I hepatic drug metabolism following intoxication by carbon tetrachloride (CCl4). Male Sprague Dawley rats (220-250 g) were divided into two sets, one included rats with or without CCl4 intoxication, the other included CCl4 intoxicated rats with or without treatment of Schisandra extract. With the treatment regimen, rats received four oral doses of Schisandra (160 mg/kg) or the same volume of water at 8, 24, 32 and 48 h after CCl4 intoxication. A single oral dose (80 mg/kg) of antipyrine, a conventional probe for oxidative drug metabolism, was then administered. The levels of liver serum transaminases and cytochrome P450 were measured and the pharmacokinetics of antipyrine were assessed using a non-compartmental approach via WinNonlin. In comparison to the rats without CCl4 intoxication (t1/2: 2.2 +/- 0.9 h; Cl/F: 0.30 +/- 0.01 L/h/Kg; P450: 0.611 +/- 0.190 nmol/mg protein), CCl4 administration significantly decreased elimination (t1/2: 12.0 +/- 3.9 h) and oral clearance (Cl/F: 0.049 +/- 0.018 L/h/kg) of antipyrine, and markedly reduced the content of P450 (0.075 +/- 0.011 nmol/mg protein). Data obtained from intoxicated animals treated by Schisandra extract, compared to those without treatment, showed significant (p < 0.05) improvement in the t1/2 (4.45 +/- 1.7 h) and Cl/F (0.096 +/- 0.018 ml/h) estimates of antipyrine and a 2-3 fold increase in P450 level (0.190 +/- 0.072 nmol/mg protein). Findings in this study suggest that the seed extract of Schisandra appeared to be a promising agent for the improvement of Phase I oxidative metabolism in the liver damaged by CCl4.

Animals↗

Effect of a lignan-enriched extract of Schisandra chinensis on aflatoxin B1 and cadmium chloride-induced hepatotoxicity in rats.

Treatment of rats with a lignan-enriched extract of the fruit of Schisandra chinensis could enhance hepatic antioxidant/detoxification system, as indicated by increases in hepatic reduced glutathione (GSH) level as well as hepatic glutathione reductase and glutathione S-transferase activities. The hepatoprotective action was evident after aflatoxin beta 1 or cadmium chloride (Cd) challenge. Schisandra chinensis pretreatment protected against aflatoxin B1-or Cd-induced hepatocellular damage in rats. However, pretreating rats with alpha-tocopherol acetate (vitamin E) did not protect against hepatic damage induced by both toxins. Results from the present as well as our previous studies demonstrate that the hepatoprotection afforded by Schisandra chinensis pretreatment is not hepatotoxin specific. Schisandra chinensis seems to be more effective than vitamin E in protecting against aflatoxin B1 and Cd toxicity. The mechanism of hepatoprotection afforded by Schisandra chinensis pretreatment may involve facilitation of both antioxidant and detoxification processes in the liver.

Aflatoxin B1↗

Identification and characterization of potent CYP3A4 inhibitors in Schisandra fruit extract.

Schisandra fruit, a Schisandraceae family herb, is used as a component in Kampo medicines (developed from Chinese medicines, but established in Japan). It can act as a sedative and antitussive, improve hepatic function, and give a general tonic effect. An extract of Schisandra fruit has been shown with a potent inhibitory effect on human liver microsomal erythromycin N-demethylation activity mediated by cytochrome P450 3A4 (CYP3A4). The present study was conducted to identify Schisandra fruit components having inhibitory effects on CYP3A4 by surveying the effect on human liver microsomal erythromycin N-demethylation activity. Known components of Schisandra fruit, gomisins B, C, G, and N and gamma-shizandrin, showed inhibitory effects on N-demethylation activity. Among these components, gomisin C displayed the most potent and competitive inhibitory effect, with a Ki value of 0.049 microM. Furthermore, the inhibitory effect of gomisin C was stronger than that of ketoconazole (Ki = 0.070 microM), a known potent CYP3A4 inhibitor. Gomisin C, however, inhibited CYP1A2-, CYP2C9-, CYP2C19-, and CYP2D6-dependent activities only to a limited extent (IC50 values >10 microM). Moreover, gomisin C inactivated human liver microsomal erythromycin N-demethylation activity in a time- and concentration-dependent manner. The inactivation kinetic parameters k(inact) and K(I) were 0.092 min(-1) and 0.399 microM, respectively. The human liver microsomal erythromycin N-demethylation activity inactivated by gomisin C did not recover on dialysis of the microsomes. Spectral scanning of CYP3A4 with gomisin C yielded an absorbance at 455 nm, suggesting that gomisin C inactivated the cytochrome P450 via the formation of a metabolite intermediate complex. This pattern is consistent with the metabolism of the methylenedioxy substituent in gomisin C. These results indicate that gomisin C is a mechanism-based inhibitor that not only competitively inhibits but irreversibly inactivates CYP3A4.

Anti-Bacterial Agents↗

Does a kampo medicine containing schisandra fruit affect pharmacokinetics of nifedipine like grapefruit juice?

Herb-drug interaction has attracted attention as medicinal topics recently. However, the drug information is sometimes confusing. Previous in vitro studies revealed that schisandra fruit had strong inhibitory effect on CYP3A4 and claimed the possibilities of its herb-drug interaction. In the present study, we evaluated the inhibitory effects of schisandra fruit and shoseiryuto, an herbal formula in Japanese traditional kampo medicine containing eight herbal medicines including schisandra fruit, on rat CYP3A activity in vitro, and the effect of shoseiryuto on pharmacokinetics of nifedipine in rats, in comparison with those of grapefruit juice, a well-characterized natural CYP3A inhibitor. Shoseiryuto and its herbal constituents, schisandra fruit, ephedra herb and cinnamon bark exhibited in vitro inhibitory effect of CYP3A. Although shoseiryuto inhibited rat CYP3A activity in vitro with a degree comparable to grapefruit juice, shoseiryuto did not significantly affect a plasma concentration profile of nifedipine in rats as grapefruit juice did. These results indicate that in vivo experiments using the extract of herbal medicine prepared with the same dosage form as patients take are necessary to provide proper information about herb-drug interaction.

Animals↗

Dibenzocyclooctadiene lignans from Schisandra chinensis protect primary cultures of rat cortical cells from glutamate-induced toxicity.

A methanolic extract of dried Schisandra fruit (Schisandra chinensis Baill.; Schisandraceae) significantly attenuated the neurotoxicity induced by L-glutamate in primary cultures of rat cortical cells. Five dibenzocyclooctadiene lignans (deoxyschisandrin, gomisin N, gomisin A, schisandrin, and wuweizisu C) were isolated from the methanolic extract; their protective effects against glutamate-induced neurotoxicity were then evaluated. Among the five lignans, deoxyschisandrin, gomisin N, and wuweizisu C significantly attenuated glutamate-induced neurotoxicity as measured by 1). an inhibition in the increase of intracellular [Ca(2+)]; 2). an improvement in the glutathione defense system, the level of glutathione, and the activity of glutathione peroxidase; and 3). an inhibition in the formation of cellular peroxide. These results suggest that dibenzocyclooctadiene lignans from Schisandra chinensis may possess therapeutic potential against oxidative neuronal damage induced by excitotoxin.

Animals↗

Separation and determination of lignans from seeds of Schisandra species by micellar electrokinetic capillary chromatography using ionic liquid as modifier.

In this paper, a micellar electrokinetic chromatographic (MEKC) method using ionic liquid as modifier for the quantification of the active components of lignans found in the medicinal herbs Schisandra species was developed for the first time. Preliminary investigations employing sodium dodecyl sulfate (SDS) as surfactant did not lead to the necessary resolution of the studied compounds, the addition of ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM-BF4) to the SDS micellar system resulted in the complete separation of all the compounds. The effects on the separation by several parameters such as BMIM-BF4 and SDS concentration, applied voltage, background electrolyte pH and concentration, were evaluated. Under the optimal conditions (5 mM borate-5 mM phosphate buffer in the presence of 20 mM SDS and 10 mM BMIM-BF4, pH 9.2, applied voltage 25 kV and detection at 254 nm), the method successfully applied to the determination of lignans in extracts of Schisandra chinensis (Turcz.) Baill. and Schisandra henryi C.B. Clarke in less than 13 min. The separation mechanism was also discussed.

Borates↗

Sedative and hypnotic activities of the ethanol fraction from Fructus Schisandrae in mice and rats.

Fruits of Fructus Schisandrae have been used as medicine for the treatment for insomnia in traditional Chinese medicine. In the present research, the sedative and hypnotic activities of the ethanol fraction of Fructus Schisandrae fruit (SY3) were studied in mice and rats. In the open field test, SY3 (25, 50 and 100 mg/kg) significantly inhibited the motor activity of mice compared to the normal. Results also showed SY3 potentiated pentobarbital-induced sleep by not only increasing the number of falling asleep and prolonging sleeping time but also reducing sleep latency. Furthermore, sleep-wake stages of rats were evaluated by polytrophic recording for 3 h after treatment. The results demonstrated that SY3 at doses of 20, 40 and 80 mg/kg behaved remarkable action on sleep architecture of rats, which contain the increase of total sleeping time, the rate of deep slow wave sleep (SWS) and mean episode duration of deep SWS, and the decrease of the latency of deep SWS. Therefore, these results suggest that the ethanol fraction of Fructus Schisandrae fruit possesses potent sedative and hypnotic activity, which supported its therapeutic use for insomnia.

Animals↗

[Determination of lignans of Schisandra medicinal plants by HPLC].

OBJECTIVE: To determinate the contents of lignans in the stems, roots and fruits of Schisandra medicinal plants. METHOD: HPLC was applied to determine the contents of schisandrin, gomisin A, schisantherin A, deoxyschizandrin, d-epigalbacine, (+)-anwulignan, wuweizisu B, 6-O-benzoylgomisin O and wuweizisu C in the stems, roots and fruits of Schisandra. RESULT: The percent contents of 9 lignans in the fruits of 5 species were 0.52% - 1.96%, and 0.02% - 1.51% in the stems of 11 species. The lignans in the fruits of S. micrantha were similar to that of S. sphenanthera, especially with high content of deoxyschizandrin. The lignans of S. viridis were similar to that of S. chinensis and with high content of schisandrin, gomisin A and wuweizisu B. (+)-Anwulignan was present in the fruits of S. henryi and S. sphenanthera, in the stems of S. micrantha and S. pubescens var. pubinervis, and in the roots of S. bicolor with percentages of 0.77%, 0.42%, 0.50%, 0.26% and 0.38% respectively. d-Epigalbacine was present in the stems of S. pubescens var. pubinervis, S. pubescens and S. glaucescens with percentages of 0.89%, 1.51% and 0.17% respectively. CONCLUSION: The lignans contents in the fruits of Schisandra were higher than that in the roots and stems. The fruits of S. henryi and S. sphenanthera, the stems of S. micrantha and S. pubescens var. pubinervis and the roots of S. bicolor, the stems of S. pubescens var. pubinervis, S. pubescens and S. glaucescens may be served as the resources plants of (+)-anwulignan and d-epigalbacine respectively.

Chromatography, High Pressure Liquid↗

In vivo antioxidant action of a lignan-enriched extract of Schisandra fruit and an anthraquinone-containing extract of Polygonum root in comparison with schisandrin B and emodin.

The in vivo antioxidant action of a lignan-enriched extract of the fruit of Schisandra chinensis (FS) and an anthraquinone-containing extract of the root of Polygonum multiflorum (PME) was compared with their respective active constituents schisandrin B (Sch B) and emodin by examining their effect on hepatic mitochondrial glutathione antioxidant status in control and carbon tetrachloride (CCl 4 )-intoxicated mice. FS and PME pretreatments produced a dose-dependent protection against CCl 4 hepatotoxicity, with the effect of FS being more potent. Pretreatment with Sch B, emodin or alpha-tocopherol (alpha-Toc) also protected against CCl 4 hepatotoxicity, with the effect of Sch B being more potent. The extent of hepatoprotection afforded by FS/Sch B and PME/emodin pretreatment against CCl 4 toxicity was found to correlate well with the degree of enhancement in hepatic mitochondrial glutathione antioxidant status, as evidenced by increases in reduced glutathione level and activities of glutathione reductase, glutathione peroxidase as well as glutathione S-transferases, in both control and CCl 4 -intoxicated mice. alpha-Toc, which did not enhance mitochondrial glutathione antioxidant status, seemed to be less potent in protecting against CCl 4 hepatotoxicity. The ensemble of results indicates that FS/PME produced a more potent in vivo antioxidant action than alpha-Toc by virtue of their ability to enhance hepatic mitochondrial glutathione antioxidant status and that the differential potency of FS and PME can be attributed to the difference in in vivo antioxidant potential between Sch B and emodin. Abbreviations. ALT:alanine aminotransferases CCl 4 :carbon tetrachloride FS:lignan-enriched extract of Schisandra fruit GRD:glutathione reductase GSH:reduced glutathione GSH-Px: Se-glutathione peroxidase GST:glutathione S-transferases mt:mitochondrial MDA:malondialdehyde PME:anthraquinone-containing fraction of Polygonum root Sch B:schisandrin B SDH:sorbitol dehydrogenase alpha-Toc:alpha-tocopherol

Alanine Transaminase↗

Effects of the Schisandra fructus water extract on cytokine release from a human mast cell line.

Schisandra fructus has been used for treatment of cough and thirst in Korea. However, its therapeutic mechanisms remain largely unclear. To investigate the biological effect of Schisandra fructus water extract (SFWE), we examined the effect of SFWE on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced pro-inflammatory cytokine secretion in the human mast cell line HMC-1. HMC-1 cells were stimulated with PMA plus A23187 in the presence or absence of SFWE. Tumor necrosis factor (TNF)-alpha, interleukin 6 (IL-6), and granulocyte-macrophage colony-stimulating factor (GM-CSF) productions were measured by enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. Inhibitory IkappaB/nuclear factor-kappaB (NF-kappaB) expression was assessed by western blot. SFWE suppressed PMA plus A23187-induced TNF-alpha, IL-6, and GM-CSF production in dose-dependent manners. Furthermore, SFWE inhibited IkappaB degradation and NF-kappaB nuclear translocation. These results suggest that SFWE inhibits the secretion of pro-inflammatory cytokines in HMC-1 cells through blockade of IkappaB degradation and NF-kappaB activation. Taken together, these findings may help elucidate the mechanism of action of this medicine in the modulation of mast cell activation in inflammatory conditions.

Calcimycin↗

[Studies on chemical constituents of Schisandra propinqua (Wall.) Hook. f. et Thoms].

OBJECTIVE: To isolate and characterize compounds from the stems of Schisandra propinqua. METHOD: Extracting with solvent, isolating by column chromatography and identifying by the spectroscopic methods. RESULT: Six dibenzocyclooctadiene lignans were isolated and identified as tigloylgomisin P(1), angeloylgomisin O(2), angeloylisogomisin O(3), kadsulignan L(4), (+/-) 5,8-epoxyl-6, 7-dimethyl-2',3',2",3"-dimethylenedioxy-4', 1"-dimethyl-1,2:3,4-dibenzo-1, 3-cyclooctadiene(5) and wuweizisu C(6). CONCLUSION: Compounds 4 and 5 were the first two dibenzocyclooctadiene lignans with an 6,9-epoxy bridge cycle discovered in the genus Schisandra. The others were originally isolated from S. propinqua.

Cyclooctanes↗

[Studies on extraction process of Fructus Schisandrae chinensis and Fructus Ligustri lucidi in gandening tablet].

OBJECTIVE: To optimize the combined extraction methods of Fructus Schisandrae Chinensis and Fructus Ligustri Lucidi. METHOD: The extraction effects of three processes (SFE CO2, alcohol-extraction, water-extraction) were compared by the contents of deoxyschizandrin, the dried extraction yield and production feasibility. RESULT: The method of alcohol-extraction was the best of the three methods, and the extraction conditions were optimized by orthogonal design as follows: sixfold 90% alcohol for two times and each for one hour. CONCLUSION: The extraction method is steady and feasible, which can be used for reference for the extraction process of Fructus Schisandrae Chinensis and Fructus Ligustri Lucidi.

Alcohols↗

Application of capillary electrochromatography using macroporous polyacrylamide columns for the analysis of lignans from seeds of Schisandra chinensis.

Capillary electrochromatography (CEC) using polymer-based monolithic stationary phase has been developed as a promising method for the determination of lignans of Schisandra chinensis. The columns were prepared by in situ copolymerisation of acrylamide, N,N'-methylenebisacrylamide, vinylsulfonic acid and lauryl acrylate in presence of poly(ethylene glycol) as a porogenic agent. The columns [33 cm (24.5 cm effective length) x 75 microm I.D.] were successfully used to analyse and quantify the major lignans in extract of the seeds of Schisandra chinensis. Good separations were achieved in less than 35 min. The calibration graphs were linear in the range 0.025-1.0 mg/ml of given lignan with correlation coefficients between 0.9951 and 0.9996. The inter-day reproducibility of the peak area were below 3.9% and the inter-day reproducibility of the migration time were below 4.2%. The results of quantitative CEC analyses were compared with those obtained by reversed-phase HPLC, the levels of schizandrin, gomisin A, gomisin N and wuweizisu C determined by CEC were in a good agreement with those determined by HPLC.

Acrylic Resins↗

Schisandra chinensis-dependent myocardial protective action of sheng-mai-san in rats.

Sheng-Mai-San (SMS), a traditional Chinese formulation used for the treatment of coronary heart disease, is comprised of Radix Ginseng, Fructus Schisandrae and Radix Ophiopogonis. Pretreatment with a lignan-enriched SMS (17 g/kg/day x 3, p.o.) was found effective in protection against isoproterenol-induced myocardial injury in rats, and in ischemia-reperfusion injury in isolated perfused hearts prepared from pretreated animals. Results obtained from pretreatment studies using extracts prepared by mixing various combinations of the three component herbs indicate that the major myocardial protective component in SMS is the lignan-enriched extract of Fructus Schisandrae.

Animals↗