Analysis of Proanthocyanidins in Plant Materials Using Hydrophilic Interaction HPLC-QTOF-MS.

Proanthocyanidins (PACs) have been proven to possess a wide range of biological activities, but complex structures limit their study of structure-function relationships. Therefore, an efficient and general method using hydrophilic interaction high-performance liquid chromatography coupled with high-resolution quadrupole time-of-flight tandem mass spectrometry (HILIC-QTOF-MS) was established to analyze PACs from different plant materials. This method was successfully applied to characterize PACs from Chinese bayberry (Myrica rubra Sieb. et Zucc.) leaves (BLPs), sorghum testa (STPs) and grape seeds (GSPs). BLPs with the degree of polymerization (DP) from 1 to 8 were separated. BLPs are mainly B-type prodelphinidins and A-type BLPs were first found in this study. STPs and GSPs belonging to procyanidins showed DP from 3 to 11 and 2 to 12, respectively. A-type linkages were found for every DP of STPs and GSPs, which were first found. These results showed that HILIC-QTOF-MS can be successfully applied for analyzing PACs from different plant materials, which is necessary for the prediction of their potential health benefits.

Mainly Dimers and Trimers of Chinese Bayberry Leaves Proanthocyanidins (BLPs) are Utilized by Gut Microbiota: In Vitro Digestion and Fermentation Coupled with Caco-2 Transportation.

Chinese bayberry leaf proanthocyanidins (BLPs) are Epigallocatechin gallate (EGCG) oligomers or polymers, which have a lot of health-promoting activity. The activity is closely related to their behavior during in vitro digestion, which remains unknown and hinders further investigations. To clarify the changes of BLPs during gastrointestinal digestion, further research is required. For in vitro digestion, including gastric-intestinal digestion, colon fermentation was applied. Caco-2 monolayer transportation was also applied to investigate the behavior of different BLPs with different degrees of polymerization. The trimers and the tetramers were significantly decreased during in vitro gastric-intestinal digestion resulting in a significant increase in the content of dimers. The dimers and trimers were the main compounds utilized by gut microbiota and they were assumed not to degrade through cleavage of the inflavan bond. The monomers and dimers were able to transport through the Caco-2 monolayer at a rate of 10.45% and 6.4%, respectively.

Rethinking the Mechanism of the Health Benefits of Proanthocyanidins: Absorption, Metabolism, and Interaction with Gut Microbiota.

Proanthocyanidins, as the oligomers or polymers of flavan-3-ol, are widely discovered in plants such as fruits, vegetables, cereals, nuts, and leaves, presenting a major part of dietary polyphenols. Although proanthocyanidins exert several types of bioactivities, such as antioxidant, antimicrobial, cardioprotective, and neuroprotective activity, their exact mechanisms remain unclear. Due to the complexity of the structure of proanthocyanidins, such as their various monomers, different linkages and isomers, investigation of their bioavailability and metabolism is limited, which further hinders the explanation of their bioactivities. Since the large molecular weight and degree of polymerization limit the bioavailability of proanthocyanidins, the major effective site of proanthocyanidins is proposed to be in the gut. Many studies have revealed the effects of proanthocyanidins from different sources on changing the composition of gut microbiota based on in vitro and in vivo models and the bioactivities of their metabolites. However, the metabolic routes of proanthocyanidins by gut microbiota and their mutual interactions are still sparse. Thus, this review summarizes the chemistry, absorption, and metabolic pathways of proanthocyanidins ranging from monomers to polymers, as well as the mutual interactions between proanthocyanidins and gut microbiota, in order to better understand how proanthocyanidins exert their health-promoting functions.

Unveiling the molecular mechanisms of Dendrobium officinale polysaccharides on intestinal immunity: An integrated study of network pharmacology, molecular dynamics and in vivo experiments.

Intestinal immunity plays a pivotal role in overall immunological defenses, constructing mechanisms against pathogens while maintaining balance with commensal microbial communities. Existing therapeutic interventions may lead to drug resistance and potential toxicity when immune capacity is compromised. Dendrobium officinale, a traditional Chinese medicine, contains components identified to bolster immunity. Employing network pharmacology strategies, this study identified constituents of Dendrobium officinale and their action targets in the TCMSP and Swiss Target Prediction databases, and compared them with intestinal immunity-related targets. Protein-protein interaction networks revealed the core targets of Dendrobium officinale polysaccharides, encompassing key pathways such as cell proliferation, inflammatory response, and immune reactions, particularly in association with the Toll-like receptor 4. Molecular docking and molecular dynamics simulation further confirmed the high affinity and stability between Dendrobium officinale polysaccharides and Toll-like receptor 4. In vivo experiments demonstrated that Dendrobium officinale polysaccharides modulates the expression of Toll-like receptor 4 and its downstream key proteins in the colonic mucosa of mice. Consequently, these findings suggest that Dendrobium officinale polysaccharides may serve as a potential modulator for intestinal immune functions, with its mechanism potentially related to the Toll-like receptor 4.

Dendrobium officinale Xianhu 2 polysaccharide helps forming a healthy gut microbiota and improving host immune system: An in vitro and in vivo study.

Dendrobium officinale is widely consumed owing to its numerous beneficial effects. We aimed to characterize polysaccharides of Dendrobium officinale (DOP) from the stems of Dendrobium officinale Xianhu 2 and clarify whether it benefit the intestinal microbiota and the immune system. The DOP weighed 291 kDa and comprised mannose, glucose, galactose, and rhamnose at 59.31:33.31:1.00:0.51 M ratio. In in vitro/vivo studies, DOP significantly increased benign intestinal microbe proportion (Lactobacillus, etc.), but reduced harmful bacteria (Escherichia_Shigella) (P < 0.05), and significantly increased butyric acid production (P < 0.05). Concentrations of 2 g/L DOP for in vitro fermentation and 100 mg/kg body weight for the mouse model were effective. In mice, DOP significantly reduced CRP, CD3, CD4, and TNF-α levels and increased C4 levels (P < 0.05). DOP might influence the immune system indirectly through regulation of the gut microbiota. Its possible regulation mechanism was that DOP reduced CD4+ Th cells proliferation so that reduced the secretion of TNF-α.

Extraction and identification of proanthocyanidins from the leaves of persimmon and loquat.

Proanthocyanidins is flavan-3-ol polymers with many activities which attracted a lot of attention. However, most of the proanthocyanidins come from fruits and seeds, resulting in higher costs. The extraction of proanthocyanidins from leaves that were trimmed as wastes from fruit trees is of good economic benefits. The proanthocyanidins in persimmon leaves and loquat leaves were extracted and purified. The purity of persimmon and loquat leaves were 85.33 ± 0.11% and 88.45 ± 0.96% with yield of 3.40% and 2.37% respectively. Detailed structure information was analyzed. Persimmon leaves proanthocyanidins mainly consist of catechin with B-type link along with a small portion of gallocatechin, catechin gallate and A-type link. Loquat leaves proanthocyanidins consist of catechin, gallocatechin, gallocatechin gallate and afzelechin with B-type link along with a small portion of A-type link. The α-amylase inhibition effect of the two leaves was analyzed. Persimmon leaves proanthocyanidins and loquat leaves proanthocyanidins were two mixed-type inhibitors to α-amylase.

Dendrobium officinale Polysaccharides Better Regulate the Microbiota of Women Than Men.

Dendrobium officinale is widely used as a health supplement, but its specific impact on healthy gut microbiota has not yet been clarified, nor has its impact on different human genders. To overcome the problems mentioned above. DOP was extracted and purified with an 8000-12,000 Da dialysis bag. The molecular weight and monosaccharide composition were determined using HPGPC and GC. Gas chromatography was used to detect the content of SCFA. 16S rDNA sequencing was used to analyze the diversity of human microbiota. The results showed that DOP contained two fractions, with an average molecular weight of 277 kDa and 1318 Da, and mainly composed of mannose and glucose. DOP can increase the relative abundance of benign microbiota and decrease the harmful types. Propionic acid content in women was significantly increased after DOP treatment. Finally, the correlation analysis revealed that DOP was beneficial to the microbiota of both men and women. It can be concluded from the results that DOP is a health supplement suitable for humans, and especially women.

Oral Administration of Fucosylated Chondroitin Sulfate Oligomers in Gastro-Resistant Microcapsules Exhibits a Safe Antithrombotic Activity.

Fucosylated chondroitin sulfate (FCS) polysaccharide isolated from sea cucumber has potent anticoagulant activity. Based on its resistance to the enzymes present in vertebrates, it may serve as an anticoagulant and shows antithrombotic effects when delivered through gastro-resistant (GR) tablets. However, due to the multiple plasma targets of FCS polysaccharide in the coagulation pathway, bleeding can occur after its oral administration. In the current study, we used FCS oligomers, in particular a mixture of oligosaccharides having 6 to 18 saccharide units, as the active ingredient in GR microcapsules for oral anticoagulation. In a Caco-2 model, the FCS oligomers showed higher absorption than native FCS polysaccharides. Oral administration of FCS oligomer-GR microcapsules provided a dose-dependent, prolonged anticoagulant effect with a selective inhibition of the intrinsic coagulation pathway when compared with subcutaneous administration of FCS oligomers or oral administration of unformulated FCS oligomers or native FCS-GR microspheres. Continued oral administration of FCS oligomer-GR microcapsules did not result in the accumulation of oligosaccharides in the plasma. Venous thrombosis animal models demonstrated that FCS oligomers delivered via GR microcapsules produced a potent antithrombotic effect dependent on their anticoagulant properties in the plasma, while oral administration of unformulated FCS oligomers at the same dose exhibited a weaker antithrombotic effect than the formulated version. Oral administration of FCS oligomer-GR microcapsules resulted in no bleeding, while oral administration of native FCS-GR microcapsules resulted in bleeding (p < 0.05). Our present results suggest that a FCS oligomer-GR microcapsule formulation represents an effective and safe oral anticoagulant for potential clinical applications.

Evaluation of antimicrobial and antibiofilm properties of proanthocyanidins from Chinese bayberry (Myrica rubra Sieb. et Zucc.) leaves against Staphylococcus epidermidis.

Staphylococcus epidermidis has emerged in recent years as one of the most important opportunistic pathogens owing to its ability to attach to processing surfaces in the food industry. Demands of foodstuffs maintaining microbiological safety and stability enhance the need to develop natural antimicrobial agents as food preservatives. Proanthocyanidins from Chinese bayberry leaves (BLPs) belonging to the class of polyphenols promise to be a potential antibacterial material against bacterial adhesion and biofilm formation. The aim of the present study was to investigate the effects of BLPs on S. epidermidis growth and biofilm formation. BLPs possessed antimicrobial activity with MIC and MBC of 320 and 640 µg/ml, respectively. Scanning electron microscopy, transmission electron microscopy, and flow cytometry analysis revealed a loss of the cell structure and function after treatment of BLPs, evidenced by cell membrane hyperpolarization and changes in cellular morphology. BLPs inhibited the biofilm formation by S. epidermidis on polystyrene microplates. Atomic force microscopy analysis showed that BLPs could decrease the stiffness and adhesion force of the cell envelope, which might account for the inhibition of biofilm formation. In summary, this study indicated that BLPs have potential to be developed as natural preservatives to control S. epidermidis in foods.

Assembly of Oil-Based Microcapsules Coated with Proanthocyanidins as a Novel Carrier for Hydrophobic Active Compounds.

Proanthocyanidins are sustainable materials with amphiphilic characteristic, network-forming capacity, and health benefits, which give them possibility as encapsulating biomaterials. We found that proanthocyanidins from Chinese bayberry leaves and grape seeds (BLPs and GSPs) were able to encapsulate oil to form spherical microcapsules of controlled size and architecture. Microcapsules encapsulated with BLPs and GSPs (BMs and GMs) exhibited different physical stability when subjected to environmental stresses. BMs showed higher physical stability to environmental stresses than GMs. The proanthocyanidin shell could protect ß-carotene from chemical degradation. Subsequently, varied gastrointestinal behaviors of the microcapsules were observed in simulated digestion. GMs with low stability reduced the lipid digestion and ß-carotene bioaccessibility. BMs with high stability retarded lipid digestion but did not change the amount of hydrolyzed lipids and ß-carotene bioaccessibility. Our study demonstrates that BLPs rather than GSPs can be used alone as encapsulating material for protection and targeted delivery of lipophilic bioactive compounds.

Fabrication of Polydopamine-Based Curcumin Nanoparticles for Chemical Stability and pH-Responsive Delivery.

Polydopamine (PDA) possesses high aqueous dispersibility, strong optical absorption, and a zwitterionic property, which give it multitudes of advantages to coat light-sensitive hydrophobic curcumin (Cur) for pH-responsive release. However, PDA is formed in alkaline conditions, which hinders its potential application for alkali-sensitive curcumin coating. Here, we developed a method to prepare PDA-coated Cur nanoparticles (NPs), which reduced chemical degradation of Cur in alkaline conditions. Encapsulation efficiency and loading capacity decreased to 73.69% and 51.80%, as the time for dopamine polymerization went on. PDA could protect Cur from light-induced degradation in powder and solution forms. Controlled release and pH-responsive delivery of PDA-coated Cur were observed under stomach and intestinal conditions compared to free Cur, which resulted from the coverage and thickness of the PDA shell and the electrostatic attraction between PDA and Cur. PDA-coated Cur NPs could be a promising way for the application of Cur in the beverage and food industry.

Preparation of a novel emulsifier by self-assembling of proanthocyanidins from Chinese bayberry (Myrica rubra Sieb. et Zucc.) leaves with gelatin.

A physicochemically stable emulsion was developed by using a novel emulsifier, which was self-assembled colloidal complex of gelatin (GLT) and proanthocyanidins from Chinese bayberry (Myrica rubra Sieb et Zucc.) leaves (BLPs), with epigallocatechin-3-O-gallate (EGCG) as structure units. The GLT-BLP colloidal complexes were spherically shaped by transmission electron microscope (TEM). The data of Fourier transform infrared spectrum (FTIR), circular dichroism (CD), isothermal titration calorimetry (ITC) revealed that the main binding force between GLT and BLPs of the colloidal complexes was hydrogen bond. The incorporation of BLPs to GLT provided GLT with stronger affinity at oil-water interface and thus enhanced the physical stability of GLT-stabilizing emulsion. In addition, the emulsions stabilized by the colloidal complexes showed higher oxidation stability than that stabilized by free GLT only. The novel emulsifier developed in this study have potential applications as functional emulsifiers in food-grade emulsions with high anti-oxidation activity.

Flavonoids, phenolic acids, carotenoids and antioxidant activity of fresh eating citrus fruits, using the coupled in vitro digestion and human intestinal HepG2 cells model.

With in-vitro digestion and human intestinal HepG2 cells, we analyzed the bioaccessibility and cell uptake of phytochemicals and determined the cellular antioxidant capacity (CAA) of fresh eating citrus fruits. The results showed that CAA of citrus fruits was higher in digesta than in extracts, and the CAA is strongly correlated with naringenin and beta-carotene uptake (p < 0.05). During in vitro digestion, vanillic acid and p-coumaric decreased, and ferulic acid increased in all citrus fruits significantly (p < 0.05); other phytochemicals varied among the fruits. During uptake, hydroxybenzoic acids, hesperidin, narirutin, naringenin and neohesperidin were detected in cells, Zeaxanthin, lutein, beta-cryptoxanthin and beta-carotene could be detected in the citrus varieties except for pummel, but hydroxycinnamic acids and hesperitin were not detected in cells. This work provides insights into the bioaccessibility and cell uptake of phytochemicals and cellular antioxidant activity of fresh eating citrus fruits.

Structural characterization and anti-proliferative activities of partially degraded polysaccharides from peach gum.

LP100R, LP10R and LP5R were isolated from peach gum by ultrafiltration. They were identified as AG II arabinogalactans composed of mannose, rhamnose, glucuronic acid, galactose, xylose and arabinose, which had a ß-d-(1→6)-galactan backbone and were branched at O-3 and O-4. LP100R, LP10R and LP5R exist in a spherical conformation with the molecular weight of 8.50 × 104 g/mol, 4.77 × 104 g/mol and 2.40 × 104 g/mol, respectively. The binding affinities of LP fractions to galectin-3 (Gal-3) were 0.77 µM for LP100R, 2.88 µM for LP10R and 5.15 µM for LP5R, respectively. Meanwhile, an anti-proliferative assay revealed that LP100R possessed higher anti-proliferative activity against HepG2 cells (IC50, 4.5 mg/mL) and MCF-7 cells (IC50, 0.43 mg/mL) than did LP10R and LP5R, which were in accordance with their binding affinities to galectin-3. Therefore, LP fractions (especially LP100R) might exert the anti-tumor activity by directly inhibiting the Gal-3 mediated proliferation of cancer cells.

Bioactive compounds and antioxidant activity of wolfberry infusion.

An infusion of the wolfberry (Lycium barbarum L.) is a traditional Asian herbal tea. This is the most commonly consumed form of dried wolfberry worldwide, yet little scientific information on wolfberry infusions is available. We investigated the effects of making infusions with hot water on the color, the content of bioactive compounds (polysaccharides, polyphenols, flavonoids and carotenoids) and the antioxidant ability of wolfberry infusions. The contents of bioactive compounds and the antioxidant activity of a wolfberry infusion increased with increased infusion temperature and time. Total polysaccharides content (TPOC), total polyphenols (TPC), total flavonoids (TFC) and total carotenoids contents (TCC) were important for determining the antioxidant capacity of wolfberry infusions with the contribution to antioxidant activity in the order TPC > TFC > TCC > TPOC. Hierarchical cluster analysis indicated preparation conditions of 100 °C for 1~3 h, 90 °C for 2~3 h and 80 °C for 2.5~3 h were equivalent as regards the value of TPC, TPOC, TFC, TCC, FRAP, DPPH and ABTS. The results of this study suggest the length of time of making a wolfberry infusion in actual real life practice is too short and different dietary habits associated with the intake of wolfberry infusion might provide the same bioactive nutrients.