| Schisandra chinensis (Turcz.) Baill.(Schisandraceae) is a well-known medicinal plantin Traditional Chinese Medicine. The fruits have been used for centuries as an antitussive,tonic and sedative agent, and in traditional medicine to improve the liver function of patientswith viral hepatitis. In traditional Chinese medicine, the dried ripe fruits of both Schisandrasphenanthera and Schisandra chinensis have long been used as Wuweizi, even though theirchemical constituents and contents of the bioactive components are quite different. Since2000, they have been accepted as two different crude drugs, Schisandrae chinensis Fructusand Schisandrae sphenantherae Fructus, respectively, by the Chinese Pharmacopoeia. In theChinese Pharmacopoeia (2010), the quality standards of S. chinensis Fructus and S.sphenantherae Fructus were not specific and could not reflect their inherent quality. In thispaper, chemical analytical methods, fingerprint techniques and DNA molecular markerstechniques were used to study S. chinensis Fructus and S. sphenantherae Fructussystemically, in order to develop a comprehensive, scientific and effective method for theevaluation and quality control for them. Moreover, we studied the protective effect of SCEson the oxidative damage of RINm5F induced by H2O2, and explored the therapeuticmechanism of SCEs for type2diabetes.1. According to the orthogonal experimental design, the optimum extracting conditionsof lignans from the S. chinensis Fructus was showed as following: extracting solvent was90%ethanol, solid-liquid ratio was1:15, supersonic extracting time was20min andextracting thrice.2. Six lignans: schisandrin C, schisandrin B, deoxyshisandrin, schisantherin A,schisandrol B and schisandrin were isolated by silica gel chromatography from S. chinensisFructus, and their structures were elucidated on1H-NMR,13C-NMR and HPLC.3. A High Performance Liqnid Chromatography (HPLC) method has been developedfor the simultaneous determination of eight lignans, schizandrin, schisandrol B, schisantherinA, schisanhenol, anwulignan, deoxyschizandrin, schizandrin B and schizandrin C, in S.chinensis Fructus and S. sphenantherae Fructus, and20batches of samples from different areas were analyzed by the developed HPLC method. The results showed that seven lignancomponents, that is schizandrin, schisandrol B, schisantherin A, schisanhenol,deoxyschizandrin, schizandrin B and schizandrin C, exists widely in S. chinensis Fructus,and schizandrin is the highest amount component with strong activity. So it was suggestedthat the content of schizandrin and the total content of the seven lignans could be as doubleindexes for quantitative examination of S. chinensis Fructus, and the contents of schizandrinand seven lignans should be4.0and14.0m g/g, respectively, for superior drugs, and theyshould be2.9and8.0mg/g, respectively, for certified drugs. Schisantherin A, anwulignanand deoxyschizandrin were present as the main components in S. sphenantherae Fructus, itwas suggested that the total content of the three lignans could be as the indexe forquantitative examination of Fructus Schisandrae Sphenantherae, and the total content shouldbe8.0mg/g for certified drugs.4. A simple, rapid and sensitive Micellar Electrokinetic Chromatography (MEKC)method has been developed for the simultaneous quantitative analysis of eight lignans,schizandrin, schisandrol B, schisantherin A, schisanhenol, anwulignan, deoxyschizandrin,schizandrin B and schizandrin C, in S. chinensis Fructus and S. sphenantherae Fructus. Toour best knowledge, this analysis may be the first example involving the simultaneousdetection of the eight lignans in S. chinensis Fructus and S. sphenantherae Fructus by theMEKC method. Compared to published methods, the analysis time of this method wasgreatly decreased with good separation efficiency.20batches of samples from different areaswere analyzed by the developed MEKC method and HPLC method, and the results weresimilar.5. A High Performance Liqnid Chromatography (HPLC) fingerprint of S. chinensisFructus has been developed and20batches of samples from different areas were analyzed bythe developed method. According to the comparative study of fingerprints, we found10communal peaks and identify8peaks.20batches of samples were classified as3clusters bycluster analysis, A1and A2were superior and certified drugs, respectively. B was S.sphenantherae Fructus. The results were similar to HPLC method.9batches of superiordrugs were confirmed to establish the mutual model. The quality of samples were assessedby "Similarity Evaluation System for Chromatographic Fingerprint of TCM2004". It wassuggested that the Similarity should be0.950and0.850for superior drugs and certified drugs.20batches of samples from different areas were analyzed by the developed method,and the results were similar to HPLC method.6. A High Performance Capillary Electrophoresis (HPCE) fingerprint of S. chinensisFructus has been developed, and20batches of samples were analyzed by the developedmethod. They were classified to be superior drugs, certified drugs and S. sphenantheraeFructus, respectively, based on the results of cluster analysis and similarity analysis, and theresults were similar to HPLC method.7. A Infrared Spectrum (IR) fingerprint of S. chinensis Fructus has been developed, and20batches of samples were analyzed by the developed method.9batches of superior drugswere confirmed to establish the reference spectrum. The quality of samples were assessed by"OPUS" based on the results of similarity analysis. The similarity between referencespectrum and the fingerprint of superior drugs, certified drugs, S. sphenantherae Fructuswere97.33～99.64%,94.86～96.63%and83.05～90.56%, respectively. The results showedthat the IR fingerprint can provides a rapid identification and quality control method of S.chinensis Fructus.8.7batches of S. chinensis and3batches of S. sphenanthera were analyzed by RandomAmplified Polymorphic DNA (RAPD) techniques.5RAPD primers were selected from60primers, and its strip was neat, distinct and stability. Among these selected primers only1primer could identify S. chinensis and S. sphenanthera.9. In this study, we observe the effects of SCEs on the rate of cell vitality, the activity ofSOD and content of MDA in RINm5F cells and cells culture fluid injured by hydrogenperoxide (H2O2), in order to discuss the possible mechanisms of the antioxidant activity ofSCEs.1) SCEs enhanced RINm5F cells growth in a dose-dependent manner in vitro;2) SCEs could protect the RINm5F cells injured by H2O2, compared with model group,high-dose group and middle-group enhanced the cell viability significantly, the cell viabilityincreased by63.4%and45.6%, respectively. The result showed that the antioxidant activityin vitro of SCEs is in dose dependent manner, and the high-dose group is the most notable.3) The contents of MDA in RINm5F cells and cells culture fluid of high-dose group andmiddle-group were significant lower than that of model group (P﹤0.05), the result showedthat SCEs can obviously alleviate the oxidative injury of RINm5F cells. 4) The activity of SOD in RINm5F cells and cells culture fluid of high-dose group andmiddle-group were significant higher than that of model group (P﹤0.05), the result showedthat SCEs can obviously enhance the antioxidant ability of RINm5F cells. |
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