Identification of Q-markers for Schisandrae Sphenantherae Fructus in treating drug-induced liver injury based on network pharmacology, fingerprint and quantitative analysis / 中国中药杂志

This study aims to establish the ultra-performance liquid chromatography(UPLC) fingerprint and multi-indicator quantitative analysis method for Schisandrae Sphenantherae Fructus(SSF) and to screen out the potential quality markers(Q-markers) of hepatoprotection based on network pharmacology. The similarity analysis was performed using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System, which showed that the similarity of the fingerprints of 15 samples from different regions ranged from 0.981 to 0.998. Eighteen common components were identified, from which 3 differential components were selected by cluster analysis and principal component analysis. The "component-target-pathway" network was built to predict the core components related to the hepatoprotective effects. Fourteen core components were screened by network pharmacology. They acted on the targets such as AKT1, CCND1, CYP1A1, CYP3A4, MAPK1, MAPK3, NOS2, NQO1, and PTGS2 to regulate the signaling pathways of lipid metabolism and atherosclerosis, hepatitis B, interleukin-17, and tumor necrosis factor. Considering the chemical measurability, characteristics, and validity, schisantherin A, anwulignan, and schisandrin A were identified as the Q-markers. The content of schisantherin A, anwulignan, and schisandrin A in the test samples were 0.20%-0.57%, 0.13%-0.33%, and 0.42%-0.70%, respectively. Combining the fingerprint, network pharmacology, and content determination, this study predicted that schisantherin A, anwulignan, and schisandrin A were the Q-markers for the hepatoprotective effect of SSF. The results can provide reference for improving the quality evaluation standard and exploring the hepatoprotective mechanism of SSF.

Optimization of ethanol reflux extraction process of Ziziphi Spinosae Semen- Schisandrae Sphenantherae Fructus based on network pharmacology combined with response surface methodology / 中国中药杂志

The present study optimized the ethanol extraction process of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus drug pair by network pharmacology and Box-Behnken method. Network pharmacology and molecular docking were used to screen out and verify the potential active components of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus, and the process evaluation indexes were determined in light of the components of the content determination under Ziziphi Spinosae Semen and Schisandrae Sphenantherae Fructus in the Chinese Pharmacopoeia(2020 edition). The analytic hierarchy process(AHP) was used to determine the weight coefficient of each component, and the comprehensive score was calculated as the process evaluation index. The ethanol extraction process of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus was optimized by the Box-Behnken method. The core components of the Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus drug pair were screened out as spinosin, jujuboside A, jujuboside B, schisandrin, schisandrol, schisandrin A, and schisandrin B. The optimal extraction conditions obtained by using the Box-Behnken method were listed below: extraction time of 90 min, ethanol volume fraction of 85%, and two times of extraction. Through network pharmacology and molecular docking, the process evaluation indexes were determined, and the optimized process was stable, which could provide an experimental basis for the production of preparations containing Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus.

Research advances in chemical constituents and hepatoprotective effect of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus / 中国中药杂志

Schisandrae has a long history of medicinal use in China. Domestic and foreign scholars have isolated a variety of chemical constituents from Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus, including lignans, volatile oils, polysaccharides, triterpenoids, organic acids, amino acids and so on. Pharmacological studies have shown that their alcohol extracts, water extracts, lignan monomers and polysaccharides could protect liver injury and reduce enzyme ability by a variety of hepatoprotective effects such as enzyme reducing, liver protecting, and antioxidant effect. In this paper, the researches on the chemical composition, hepatoprotective effect and pharmacokinetics of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus in the past forty years were systematically collated, in order to provide useful enlightenment for the clinical application and new drug development of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus in liver protection.

Investigation on protective effect and efficacy difference of extract of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus against acetaminophen-induced liver injury / 中国中药杂志

The study was aimed to investigate the protective effect and pharmacodynamic difference of the ethanol extracts of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus on the drug-induced liver injury induced by acetaminophen.The cell activations of LO2 cells treated by Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extracts were tested by CCK-8 essay.The effects of ethanol extracts on cell survival rate,the activities of ALT and AST in culture medium were detected based on the injury model of LO2 cells induced by APAP.Further,in purpose to observe the protective effect of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extracts on a mouse model of liver injury induced by intraperitoneal injectionof acetaminophen was established.Mice were randomly divided into control group,model group,positive drug group and Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extracts administration groups.The activities of ALT and AST in the serum and the levels of MDA,SOD,GSH and GSH-PX in the liver homogenate of the mice were detected by commercial kits.The HEstaining was used to observe the histopathological changes of liver tissue in each group and the TUNEL staining was used to observe the hepatocyte apoptosis.The results showed that the ethanol extracts at less than 1 g·L~(-1)did not affect the activity of LO2 cell.Compared with the model group,the cell survival rates of the Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extract administration groups was significantly increased;the ALT and AST in the culture medium were distinct decreased(P<0.05 or P<0.01).The survival rate of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extract from different batches were similar,while that of the Schisandrea Sphenatherae Fructus ethanol extract from different batches were quite different(P<0.05or P<0.01).Further,animal experiments showed that Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extract administration groups could markedly inhibit the increase of ALT and AST levels in serum(P<0.01),decrease MDA content significantly(P<0.01),and increase GSH,GSH-PX and SOD activity significantly(P<0.01).Among them,compared with other groups,Schisandrae Sphenantherae Fructus ethanol extract-2 group showed the best effect(P<0.05 or P<0.01)while Schisandrae Sphenantherae Fructus ethanol extract-1 showed a poor effect(P<0.05 or P<0.01).In conclusion,both Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extracts have protective effect on APAP-induced drug-induced liver injury and there was a certain difference in the efficacy between Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus ethanol extracts from different habitats.

Effects of Vinegar Fructus Schisandrae Sphenantherae on Lipid Metabolism in Rats with Type 2 Diabetes / 中国中医药信息杂志

Objective To investigate the effects of Schisandrae Sphenantherae Fructus and processed with vinegar on lipid metabolism of rats with type 2 diabetes mellitus (T2DM). Methods The rat model of T2DM was induced by high fat diet plus STZ. The rats were randomly divided into blank group, model group, Schisandrae Sphenantherae Fructus high-dose group and low-dose group, and vinegar Schisandrae Sphenantherae Fructus high-dose group and low- dose group. The rats in each group were fed with the corresponding medicine for gavage for 30 d. FINS, FFA, TC, TG, HDL-C, LDL-C, and MDH, total protein content of liver tissue were detected. HE staining was used to observe the histomorphological changes of liver and pancreas in rats. Results Compared with the model group, Schisandrae Sphenantherae Fructus groups and vinegar Schisandrae Sphenantherae Fructus groups did not show obvious effects on improving FBG and FINS, but it could raise varying degrees of HDL-C and MDH, and reduce FFA, LDL-C, TC, and TG, among which vinegar Schisandrae Sphenantherae Fructus could significantly regulate metabolism in T2DM rats. Conclusion Vinegar Schisandrae Sphenantherae Fructus can enhance the lipid metabolism regulatory function of T2DM rats.

Acute Toxicity and Protective Effects of Polysaccharide from Schisandrae Sphenantherae Fructus in Mice with Liver Injury / 中国药师

Objective:To evaluate the acute toxicity and protective effects of polysaccharide from Schisandrae sphenantherae Fruc-tus in the mice with acute liver injury induced by CCl4 . Methods:The classical acute toxicity tests were used to determine the maxi-mum drug loading (MLD) and observe the acute toxicity. The acute liver injury model was induced by 0. 5% CCl4. Sixty mice were randomly divided into the blank control group, CCl4 model group, bifendatatum model group and polysaccharide group respectively at high (1388 mg·kg-1), medium (694 mg·kg-1) and low (347 mg·kg-1) dose groups with 10 ones in each. After 6-day drug treatment, the levels of aspartate aminotransferase ( AST) and alanine aminotransferase ( ALT) were determined by kits. Histopatho-logical examination was also carried out. Results: General behavior, body weight and organ examination in the acute toxicity tests showed no obvious changes. Compared with those in the model group, the levels of AST in the three polysaccharide groups significantly decreased (P<0. 05 or P<0. 01), the levels of ALT in polysaccharide group at high and medium doses significantly decreased(P<0. 01), and the liver tissue damage was improved to a certain extent as well. Conclusion: Polysaccharide from Schisandrae sphenan-therae Fructus has protective effects on acute liver injury induced by CCl4 in mice with low acute toxicity.

Analysis of commercial grades of Schisandrae Sphenantherae Fructus based on schisantherin / 中国中药杂志

Schisandrae Sphenantherae Fructus, from different producing areas, were collected and divided into three grades. The moisture content and total ash were determined on the basis of the pharmacopoeia method, and schisantherin was determined by UPLC. The study is aimed to find the commercial specifications and grades of Schisandrae Sphenantherae Fructus based on schisantherin content. The results showed that the content of water, total ash and schisantherin of Schisandrae Sphenantherae Fructus from different producing areas qualified. There was no significant difference between different grades of schisandrin content, but the second-class were the highest, first-class and third-class were lower. It means that schisandrin content is not positive correlation to commercial grade. The study will be helpful to the production, management and clinical practice of Schisandrae Sphenantherae Fructus.

Analysis of Volatile Chemical Constituents in Vinegar-Processed Schisandrae Sphenantherae Fructus and Vinegar-Processed Schisandrae Chinesis Fructus by HS-SPME-GC-MS / 中国药师

Objective:To analyze and compare the volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus.Methods:The volatile components in vinegar-processed schisandrae sphenantherae fruc-tus and vinegar-processed schisandrae chinesis fructus were extracted by headspace solid phase-microextraction (HS-SPME) and quali-tatively analyzed by GC-MS.Results:Totally 20 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.55%of the total volatile components , and totally 21 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.90% of the total volatile components .Conclusion: The type and content of volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus are quite different , and the study can provide scientific basis for the two traditional Chinese medicinal materials .

Simultaneous Determination of Five Major Lignans from Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus by Microemulsion Liquid Chromatography / 中国中医药信息杂志

Objective To develop a stable and sensitive microemulsion liquid chromatography method by using oil-in-water microemulsion mobile phase;To simultaneously determine five major active lignans, namely, schizandrin, schisandrol B, schisantherin A, deoxyschizandrin, and schisandrin B, in Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus. Methods The quantitative determination was conducted by using Kromasil C18 column (250 mm × 4.6 mm, 5μm). The influence of the composition of the microemulsion system on separation effect and retention time was studied. The method was optimized to sodium dodecyl sulfonate 3.3%(SDS, W/V), n-octanol 1.2% (V/V) and n-butanol 6.6% (V/V) in 20 mmol/L phosphate buffer (adjusted to pH 7.0 with orthophosphoric acid and sodium hydroxide). The flow rate was set at 0.7 mL/min. The compound peaks were detected by UV at 254 nm. The column temperature was 25 ℃. Results Under the optimized conditions, baseline separation of 5 lignans was achieved in 20 min, and the separation was satisfying. Schizandrin, schisandrol B, schisantherin A, deoxyschizandrin, and schisandrin B were in linear relation within the ranges of 2.0-155.0, 8.3-166.7, 8.8-175.8, 8.3-166.7, 8.9-177.1μg/mL (r>0.999). The limit of detection ranged from 0.06μg/mL to 0.18μg/mL, and the limit of quantitation ranged from 0.17μg/mL to 0.55μg/mL. The average recovery values were between 97.22%and 100.8%. Conclusion This method is simple, accurate and reliable, and can be successfully applied to simultaneous determination of five major active lignans in 13 batches of Schisandrae Sphenantherae Fructus and Schisandrae Chinensis Fructus from different producing areas in China.

Quality control study of wuziyanzong pills / 中国药学杂志

OBJECTIVE: To establish the quality standard of Wuziyanzong pills so as to improve the quality control capacity. METHODS: Thin-layer chromatography (TLC) was used for the qualitative analysis of Lycii Fructus in the formula. The fingerprint of Wuziyanzong pills based on combined control herbs, the screening method of Schisandrae Sphenantherae Fructus and the determination of hyperoside and schisandrin in the formula were established by HPLC. The contents of Pb, Cd, As, Hg, Cu and Cr were determined by ICP-MS method. RESULTS: The TLC identification method showed good separation effect and specificity. The fingerprint, using the combined control herbs, could simultaneously identify Rubi Fructus, Cuscutae Semen, Plantaginis Semen, and Schisandrae Chinensis Fructus with good method validation results. According to the peak area ratio of schisantherin A and schisandrin, it could screen out Schisandrae Sphenantherae Fructus in the formula when the ratio was proposed to be 0.20. In the assay, the content limits of hyperoside and schisandrin in Wuziyanzong pills were proposed. Three harmful elements, ie As, Hg and Pb exceeded the permitted limits in 19 batches of samples. This may be caused by using the same production line for different products and the samples may be contaminated by the residue of previous products. CONCLUSION: The established quality standard is simple and accurate and shows good specificity and reproducibility. It could be used for the comprehensive and overall quality evaluation of Wuziyanzong pills.