Research progress in the preparation, structural characterization, and biological activities of polysaccharides from traditional Chinese medicine.

Traditional Chinese medicines are tremendous sources of polysaccharides, which are of great interest in the human welfare system as natural medicines, food, and cosmetics. This review aims to highlight the recent trends in extraction (conventional and non-conventional), purification and analytic techniques of traditional Chinese medicine polysaccharides (TCMPs), and the chemical structure, biological activities (anti-tumor, hypoglycemic, antioxidant, intestinal flora regulation, immunomodulatory, anti-inflammatory, anti-aging, hypolipidemic, hepatoprotective, and other activities), and the underlying mechanisms of polysaccharides extracted from 76 diverse traditional Chinese medicines were compared and discussed. With this wide coverage, a total of 164 scientific articles were searched from the database including Google Scholar, PubMed, Web of Science, and China Knowledge Network. This comprehensive survey from previous reports indicates that TCMPs are non-toxic, highly biocompatible, and good biodegradability. Besides, this review highlights that TCMPs may be excellent functional factors and effective therapeutic drugs. Finally, the current problems and future research advances of TCMPs are also introduced. New valuable insights for the future researches regarding TCMPs are also proposed in the fields of therapeutic agents and functional foods.

Rapid Isolation and Hypoglycemic Activity of Secondary Metabolites of Eurotium cristatum by High-Speed Countercurrent Chromatography.

In this study, secondary metabolites of Eurotium cristatum were isolated and purified by high-speed counter-current chromatography (HSCCC), and their hypoglycemic activities were studied. The general-useful estimate of solvent systems (GUESS) for counter-current chromatography was employed to select the appropriate solvent systems of n-hexane-ethyl acetate-methanol-water (HEMW, 4:6:5:5, v/v/v/v) for HSCCC practice, and three compounds were separated from the crude ethyl acetate extract of E. cristatum in one single step; 6.1 mg of Compounds 1, 5.6 mg of Compound 2 and 3.8 mg of Compound 3 were obtained from 100 mg of crude extract with a stationary phase retention of 75%. The compounds were then identified as emodin methyl ether, chrysophanol and emodin, respectively. The activity of the target compounds in the secondary metabolites of E. cristatum was verified by testing their inhibition on α-glucosidase activity and molecular docking simulation. The results showed that emodin, chrysophanol and emodin methyl ether had significant inhibitory effects on the α-glucosidase activity. This work confirmed the effectiveness of HSCCC in the separation of compounds in complex extracts and provided reference for further research and application of E. cristatum.

A simple and practical solvent system selection strategy for high-speed countercurrent chromatography based on the HPLC polarity parameter model.

The selection of an appropriate solvent system is the most crucial step in high-speed countercurrent chromatography (HSCCC) separation. The compound polarity plays an important role in HPLC analysis and HSCCC separation, and it can be calculated by the HPLC polarity parameter model and the average polarity of the HSCCC solvent system, respectively. However, flow rates, columns and methanol concentrations of the HPLC experiment can influence the calculation of the compound polarity. Therefore, the applicability and accuracy of the HPLC polarity parameter model still needed to be extensively validated. We chose 14 compounds to conduct the shake-flask experiments and HPLC analysis on, such as apigenin, honokiol, phloridzin and dihydromyricetin. The HPLC analysis results showed that different flow rates and columns have negligible effects on the calculated compound polarities. However, there was a certain variation trend in the calculated polarities with different methanol concentrations. Although the polarity values of some compounds showed a difference between the HPLC analysis and shake-flask experiments, their partition coefficients (K) in the HSCCC solvent systems were still located in the range of 0.5 < K < 2.0. Guided by the HPLC polarity parameter model, the appropriate HSCCC solvent systems for mangosteen peel and Hypericum sampsonii Hance were selected, and the two main components (mangostin and quercetin) were isolated from their extracts, respectively. The separation results showed that the predicted compound polarities were sufficient to meet the HSCCC separation requirements. Meanwhile, this method required only 1 to 2 HPLC analyses with reference compounds, greatly improved the efficiency of the HSCCC solvent system selection, and shortened the experimental time. The polarity parameter model was a fast and efficient analysis method for the selection of an appropriate HSCCC solvent system.

Recent advances in medicinal and edible homologous polysaccharides: Extraction, purification, structure, modification, and biological activities.

110 kinds of traditional Chinese medicines can be used for medicine and food from Chinese pharmacopoeia in 2021. With the deepening of research in recent years, medicinal and edible homologous (MEH) traditional Chinese medicines have great development and application prospects in many fields. Polysaccharides are one of the major and representative pharmacologically active macromolecules in traditional Chinese medicines with MEH. Moreover, traditional Chinese medicines with MEH have become the main source of natural polysaccharides with safety, high efficiency, and low side effects. Increasing researches have confirmed that MEH polysaccharides (MEHPs) have multiple biological activities both in vitro and in vivo methods, such as antioxidant, immunomodulatory, anti-tumor, anti-aging, anti-inflammatory, hypoglycemic, hypolipidemic activities, and regulating intestinal flora. Additionally, different raw materials, extraction, purification, and chemical modification methods result in differences in the structure and biological activities of MEHPs. The purpose of the present review is to provide comprehensively and systematically reorganized information in the extraction, purification, structure, modification, biological activities, and potential mechanism of MEHPs to support their therapeutic effects and health functions. New valuable insights and theoretical basis for the future researches and developments regarding MEHPs were proposed in the fields of medicine and food.

Exploring the Protective Effects and Mechanism of Huaji Jianpi Decoction against Nonalcoholic Fatty Liver Disease by Network Pharmacology and Experimental Validation.

This paper was designed to predict the mechanisms of the active components of Huaji Jianpi Decoction (HJJPD) against nonalcoholic fatty liver disease (NAFLD) based on network pharmacology-combined animal experiments. The candidate compounds of HJJPD and its relative targets were obtained from TCMSP and PharmMapper web server, and the intersection genes for NAFLD were discerned using OMIM, GeneCards, and DisGeNET. Then, the target protein-protein interaction (PPI) and component-target-pathway networks were constructed. Moreover, gene function annotation (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to study the potential signaling pathways associated with HJJPD's effect on NAFLD. Molecular docking simulation was preformed to validate the binding affinity between potential core components and key targets. Eventually, the candidate targets, the possible pathway, and the mechanism of HJJPD were predicted by the network pharmacology-based strategy, followed by experimental validation in the NAFLD mice model treated with HJJPD. A total of 55 candidate compounds and 36 corresponding genes were identified from HJJPD that are associated with activity against NAFLD, and then the network of them was constructed. Inflammatory response and lipid metabolism-related signaling pathways were identified as the critical signaling pathways mediating the therapeutic effect of the active bioactive ingredients on NAFLD. Compared with the model group, the liver wet weight, liver/body ratio, the levels of total cholesterol (TC), triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and high-density lipoprotein (HDL) in serum in the HJJPD low-dose (17.52 g/kg·d), medium-dose (35.04 g/kg·d), and high-dose (70.07 g/kg·d) groups significantly decreased (P < 0.05). Light microscope observation shows that HJJPD could control the degree of lipid denaturation of the mouse liver tissue to a great extent. RT-qPCR results show that the mRNA expression levels of peroxisome proliferative activated receptor gamma (PPARG), tumor necrosis factor-α (TNF-α), antiserine/threonine protein kinase 1 (AKT1), and prostaglandin-endoperoxide synthase (PTGS2) in the liver tissues of the three HJJPD groups (17.52 g/kg·d, 35.04 g/kg·d, and 70.07 g/kg·d) were significantly lower than those in the model group (P < 0.05). HJJPD can exert its effect by inhibiting hepatic steatosis and related mRNA expression and decreasing the levels of other liver-related indexes. This study suggested that HJJPD exerted its effect on NAFLD by modulating multitargets with multicompounds through multipathways. It also demonstrated that the network pharmacology-based approach might provide insights for understanding the interrelationship between complex diseases and interventions of HJJPD.

Extraction, purification, structure, modification, and biological activity of traditional Chinese medicine polysaccharides: A review.

Traditional Chinese medicines (TCM), as the unique natural resource, are rich in polysaccharides, polyphenols, proteins, amino acid, fats, vitamins, and other components. Hence, TCM have high medical and nutritional values. Polysaccharides are one of the most important active components in TCM. Growing reports have indicated that TCM polysaccharides (TCMPs) have various biological activities, such as antioxidant, anti-aging, immunomodulatory, hypoglycemic, hypolipidemic, anti-tumor, anti-inflammatory, and other activities. Hence, the research progresses and future prospects of TCMPs must be systematically reviewed to promote their better understanding. The aim of this review is to provide comprehensive and systematic recombinant information on the extraction, purification, structure, chemical modification, biological activities, and potential mechanism of TCMPs to support their therapeutic effects and health functions. The findings provide new valuable insights and theoretical basis for future research and development of TCMPs.

Extraction of phenolics and anthocyanins from purple eggplant peels by multi-frequency ultrasound: Effects of different extraction factors and optimization using uniform design.

In this work, multi-frequency ultrasound (working modes for the single-, dual- and tri-frequency in simultaneous ways) was applied to extract bioactive compounds from purple eggplant peels. The single-factor experiments were performed by varying six independent variables. A six-level-five-factor uniform design (UD) was further employed to evaluate the interaction effects between different factors. It was found that extraction temperature and extraction time significantly affected the total phenolic content (TPC), whereas the total monomeric anthocyanins (TMA) was mainly influenced by ethanol concentration, extraction temperature and solid-liquid ratio. Based on partial least-squares (PLS) regression analysis, the optimal conditions for TPC extraction were: 53.6 % ethanol concentration, 0.336 mm particle size, 44.5 °C extraction temperature, 35.2 min extraction time, 1:43 g/mL solid-liquid ratio, and similar optimal conditions were also obtained for TMA. Furthermore, the TPC and TMA extraction were investigated by ultrasound in different frequencies and power levels. Compared with single-frequency (40 kHz) and dual-frequency ultrasound (25 + 40 kHz), the extraction yield of TPC and TMA with tri-frequency ultrasound (25 + 40 + 70 kHz) increased by 23.65 % and 18.76 % respectively, which suggested the use of multi-frequency ultrasound, especially tri-frequency ultrasound, is an efficient strategy to improve the TPC and TMA extracts in purple eggplant peels.

Polysaccharides from Medicine and Food Homology Materials: A Review on Their Extraction, Purification, Structure, and Biological Activities.

Medicine and food homology (MFH) materials are rich in polysaccharides, proteins, fats, vitamins, and other components. Hence, they have good medical and nutritional values. Polysaccharides are identified as one of the pivotal bioactive constituents of MFH materials. Accumulating evidence has revealed that MFH polysaccharides (MFHPs) have a variety of biological activities, such as antioxidant, immunomodulatory, anti-tumor, hepatoprotective, anti-aging, anti-inflammatory, and radioprotective activities. Consequently, the research progress and future prospects of MFHPs must be systematically reviewed to promote their better understanding. This paper reviewed the extraction and purification methods, structure, biological activities, and potential molecular mechanisms of MFHPs. This review may provide some valuable insights for further research regarding MFHPs.

Ultrasonic-Assisted Aqueous Two-Phase Extraction and Properties of Water-Soluble Polysaccharides from Malus hupehensis.

Malus hupehensis (M. hupehensis), an edible and medicinal plant with significant antioxidant and hypoglycemic activity, has been applied to new resource foods. However, the structural characterization and biological effects of its polysaccharides (MHP) are less known. The optimum extraction parameters to achieve the highest extraction efficiency (47.63%), the yield (1.68%) and purity of MHP (89.6%) by ultrasonic-assisted aqueous two-phase system (ATPS) were obtained under the liquid-to-solid ratio of 23 g/mL, ultrasonic power of 65 W, and ultrasonic time of 33 min. According to the analysis results, MHP was composed of Man, GlcA, Rha, GalA, Glc, Gal, Xyl, Ara, and Fuc, in which Ara and Gal were the main components, and the content of GlcA was the lowest. In in vitro activity analysis, MHP showed a significant antioxidant capacity, and an inhibition activity of α-glucosidase and the advanced glycation end products (AGEs) formation in the BSA/Glc reaction model. MHP interacted with α-glucosidase and changed the internal microenvironment of the enzyme, and inhibited the AGEs formation, which provides more evidence for the antihyperglycemic mechanism of MHP. The results suggest that ATPS is an efficient and environmentally friendly solvent system, and M. hupehensis has broad application prospects in functional foods, healthcare products, and pharmaceuticals.

Extraction and separation of flavonoids from Malus hupehensis using high-speed countercurrent chromatography based on deep eutectic solvent.

In the present experiment, a green and highly efficient extraction method for flavonoids established on deep eutectic solvents (DESs) was investigated by using the response surface methodology. The DES-based high-speed countercurrent chromatography (HSCCC) solvent systems were developed for the separation of high purity compounds from the DES extract of Malus hupehensis for the first time. Under the optimal conditions (liquid-to-solid ratio of 26.3 mL/g, water content of 25.5%, and extraction temperature of 77.5°C), the yield of flavonoids was 15.3 ± 0.1%, which was superior to that of the methanol extraction method. In accordance with the physical property of DES-based HSCCC solvent systems and K values of target compounds, DES-based HSCCC solvent systems composed of choline chloride/glucose-water-ethyl acetate (ChCl/Glu-H2O-EAC, 1:1:2, v/v) was selected for the HSCCC separation. Thus, five flavonoids (two novel compounds 1-2, 6´´-O-coumaroyl-2´-O-glucopyranosylphloretin and 3´´´-methoxy-6´´-O-feruloy-2´-O-glucopyranosylphloretin; three know compounds 3-5, namely, avicularin, phloridzin, and sieboldin) were efficiently separated from Malus hupehensis. DESs are the environment friendly and highly efficient solvents as the components of extraction solvent and HSCCC solvent system, and can be re-utilized many times. However, ethyl acetate can be soluble with a few hydrogen bond donors, such as urea, carboxylic acid and polyol, through the shake flask test. It is the great difficulty for the efficient and rapid separation of target compounds from the DESs extract because of the DESs residual in the HSCCC fractions. ChCl and Glu are the great choices of DESs without this problem. In addition, K values increased with the increase of the molar ratio of ChCl/Glu and the content of water, which could effectively guide us to choose the suitable DES-based HSCCC solvent system. The twice HSCCC separation results indicated that DES was the valuable and green solvent for the HSCCC separation of pure compounds from the extract for the first time, and showed the recycle superiority of DES-based HSCCC solvent system.

Separation of six antioxidants from Hypsizygus marmoreus by high-speed countercurrent chromatography utilizing an approach based upon the polarity parameter model.

For a successful high-speed countercurrent chromatography (HSCCC) separation of multiple components, the suitable solvent system selection is a critical operation. Despite the difference in separation mechanism between HSCCC and HPLC, K values of compounds in the solvent system and the retention factor of compounds in HPLC were connected with polarity, and the polarity mainly depended on the structure and chemical properties of compounds. On the basis of the concept of "like dissolves like", the average polarity of the solvent system is equal to the average polarity of multiple components at the "sweet point" log K = 0. The result of theoretical deduction showed a negative linear correlation between the polarity of each component and the logarithm of the gradient range of the organic phase. Therefore, the average polarity of unknown multiple components could be obtained through the polarity parameter model established in the HPLC analysis. The suitable solvent system composed of n-hexane-ethyl acetate-methanol-water (8:3:8:1, v/v) was selected through the average polarity of multi-components combining K values of the enriched samples. In the context of bioassay, an efficient HSCCC separation procedure was established, and two groups of analogue benzaldehyde derivatives (four main antioxidants, namely, isodihydroauroglaucin, isoaspergin, isotetrahydro-auroglaucin, and flavoglaucin; two minor antioxidants, namely, 6'-oxo-chaetopyranin and chaetopyranin) were obtained from Hypsizygus marmoreus. The predicted polarity values of multi-components were sufficient to meet the HSCCC experimental requirements. The HPLC analysis of reference compounds and multi-components showed a significant consistency with different chromatographic columns. Therefore, the polarity parameter model established in the HPLC analysis was a simple, rapid, and helpful tool for looking an appropriate solvent system, which was a forwarding step for the HSCCC separation of multiple components.

Ultrasound-Assisted Deep Eutectic Solvent Extraction of Anthocyanins from Blueberry Wine Residues: Optimization, Identification, and HepG2 Antitumor Activity.

Blueberry wine residues produced during the wine-brewing process contain abundant anthocyanins and other bioactive compounds. To extract anthocyanins from blueberry wine residues more efficiently, a novel procedure of ultrasound-assisted deep eutectic solvent extraction (UADESE) was proposed in this work. The extraction process was optimized by response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest yield of anthocyanins (9.32 ± 0.08 mg/g) from blueberry wine residues by UADESE were obtained at water content of 29%, ultrasonic power of 380 W, extraction temperature of 55 °C, and extraction time of 40 min. The AB-8 macroporous resin combined with Sephadex LH-20 techniques was used to purify the crude extract (CE) obtained under optimum extraction conditions and analyze the anthocyanins composition by HPLC-ESI-MS/MS. The cyanidin-3-rutinoside with purity of 92.81% was obtained. The HepG2 antitumor activity of CE was better than that of the purified anthocyanins component. Moreover, CE could increase the intracellular reactive oxygen species levels and the apoptosis, and arrest HepG2 cells in the S phases. These findings provided an effective and feasible method for anthocyanins extraction, and reduced the environmental burden of this waste.

Ultrasound-assisted enzymatic extraction of anthocyanins from grape skins: optimization, identification, and antitumor activity.

Grape skins produced during the grape juice production and processing contain abundant anthocyanins and other active compounds. Consequently, this study optimized the extraction conditions for ultrasound-assisted enzymatic extraction (UAEE) of anthocyanins from grape skins via response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest anthocyanins yield (3.01 ± 0.04) mg/g from grape skins by UAEE were obtained under an extraction temperature of 50 °C, ultrasonic power of 400 W, pectinase dosage of 0.16%, and extraction time of 28 min. The AB-8 macroporous resin combined Sephadex LH-20 techniques were further employed to purify the anthocyanins extracts obtained under optimum extraction conditions (AEOEC), and the main anthocyanins were identified using high-performance liquid chromatography tandem mass spectrometry. The purified anthocyanins contained two anthocyanins in terms of delphinidin-3,5-O-diglucoside and cyanidin-3-O-rutinoside with purity of 91.35% and 92.64%, respectively. Ultimately, we further evaluated the antitumor activity of AEOEC and two purified anthocyanins on breast cancer. The results indicated that the antitumor effect of AEOEC on breast cancer MCF-7 cells was better than that of two purified anthocyanins. In addition, AEOEC could memorably increase intracellular reactive oxygen species levels and apoptosis of MCF-7 cells, and arrest MCF-7 cells in the G2/M phases. The findings provide an effective and feasible method for anthocyanins extraction and reduce the environmental burden of this waste.

Dichloromethane Extract of Fermentation Broth by Co-Culture of Morchella esculenta and Coprinus comatus Induces Apoptosis in U251 Cells via Mitochondrial Intrinsic Pathway.

The current study was designed to investigate the inhibitory effects of dichloromethane extract of fermentation broth by co-culture of Morchella esculenta and Coprinus comatus (DCMM) on human glioma U251 cells in vitro and its possible underlying mechanisms. The proliferation of U251 cells was inhibited by DCMM with different concentrations by the CCK-8 assay. Besides, flow cytometry assay was used to evaluate the DCMM promoted U251 cell apoptosis rate in a dose-dependent manner. DCMM with different concentrations (10 µg·mL-1, 20 µg·mL-1, and 40 µg·mL-1) significantly enhanced the expression of caspases-3 activity after 24 h. In addition, DCMM with different concentrations significantly increased caspase-3 and Bax, and decreased Bcl-2 expressions at both mRNA and protein levels. DCMM can remarkably inhibit the proliferation and promote cell apoptosis of human glioma U251 cells. The possible underlying mechanisms could be related to induction of apoptosis of human glioma U251 cells by mitochondrial intrinsic pathway.