Functional Food Applications Research Articles

Research progress on extraction, purification, structure and biological activity of Dendrobium officinale polysaccharides.

Dendrobium officinale Kimura et Migo (D. officinale) is a traditional medicinal and food homologous plant that has been used for thousands of years in folk medicine and nutritious food. Recent studies have shown that polysaccharide is one of the main biologically active components in D. officinale. D. officinale polysaccharides possess several biological activities, such as anti-oxidant, heptatoprotective, immunomodulatory, gastrointestinal protection, hypoglycemic, and anti-tumor activities. In the past decade, polysaccharides have been isolated from D. officinale by physical and enzymatic methods and have been subjected to structural characterization and activity studies. Progress in extraction, purification, structural characterization, bioactivity, structure-activity relationship, and possible bioactivity mechanism of polysaccharides D. officinale were reviewed. In order to provide reference for the in-depth study of D. officinale polysaccharides and the application in functional food and biomedical research.

A Comprehensive Review of C. capsularis and C. olitorius: A Source of Nutrition, Essential Phytoconstituents and Pharmacological Activities.

Plant bioactive compounds have gained global significance in terms of both medicinal and economic ramifications due to being easily accessible and are believed to be effective with fewer side effects. Growing relevant clinical and scientific evidence has become an important criterion for accepting traditional health claims of medicinal plants and also supports the traditional uses of Corchorus as folk medicine. C. capsularis and C. olitorius have broad applications ranging from textile to biocomposite, and young leaves and shoots are used as healthy vegetables and have long been used as traditional remedies for fever, ascites, algesia, liver disorders, piles, and tumors in many cultures. This review systematically summarized and emphasized the nutritional attributes, mostly available bioactive compounds, and biological and potential pharmaceutical properties of C. capsularis and C. olitorius, disclosed to users and non-users. Results suggest that various phytochemicals such as cardiac glycosides, phenols, flavonoids, sterols, lipids, and fatty acids were found or analytically identified in different plant parts (leaf, stem, seed, and root), and many of them are responsible for pharmacological properties and their antitumor, anticancer, antioxidant, antinociceptive, anti-inflammatory, analgesic, antipyretic, antiviral, antibacterial, anticonvulsant, antidiabetic and antiobesity, and cardiovascular properties help to prevent and cure many chronic diseases. In addition to their use in traditional food and medicine, their leaves have also been developed for skin care products, and some other possible uses are described. From this review, it is clear that the isolated compounds of both species have great potential to prevent and treat various diseases and be used as functional foods. In conclusion, this comprehensive review establishes a significant reference base for future research into various medical and functional food applications.

Sour cherry juice concentrate powdered by high and low temperature spray drying with pea protein as a carrier—Physical properties, antioxidant activity and in vitro bioaccessibility

Spray drying of sour cherry juice concentrate was performed by conventional high temperature method (180 °C/80 °C, inlet/outlet air), as well as by innovative approach with dehumidified air and low drying temperature (80 °C/45 °C, inlet/outlet air). Moreover, the possibility to apply pea protein as an alternative carrier to partially replaced maltodextrin was tested. Due to lower drying temperature and reduced stickiness the carrier content was reduced from 50% down to 30% solids to enhance powders physiochemical properties (lower stickiness, hygroscopicity, better color, better bioaccessibility), although some parameters of variants produced with the addition of pea protein had poorer quality in comparison to powders with only maltodextrin (moisture content, water activity, particles morphology and size, glass transition temperature). Increased bioaccessibility of phenolics and antioxidant potential of pea protein-enriched samples indicated the potentially possible application in functional food.

The effect of psyllium (Plantago ovata Forsk) fibres on the mechanical and physicochemical characteristics of plant-based sausages.

Psyllium is a source of natural dietary fibre with recognised health benefits that can be used as a hydrocolloid with functional food applications. The purpose of this study was to determine the effect of different levels of Plantago ovata fibres in plant-based sausages on their composition, physicochemical, and mechanical properties. Proximate composition was studied. Water activity (aw), water release, pH, colour measurement, texture profile analysis (TPA), and Warner–Bratzler Shear Force (WBSF) were determined to establish the physicochemical and textural properties of sausages. A plant-based sausages microstructure study and a sensory study were carried out to better understand conformation and to determine their acceptance. The results showed that sausages had high ash and carbohydrate contents but, above all, a low-fat content. The use of psyllium increased water-holding capacity. The results also indicated that employing Plantago ovata white (PW) fibre can minimise mechanical problems and reduce colour changes. However, PW fibre showed less retained water, which was why chickpea starch further developed and was more gelatinised. At the same time, the plant-based sausages with PW fibre obtained the best overall score with the fewest colour changes in the sensory evaluation. Nevertheless, further studies are recommended to improve the texture and acceptability of these plant-based sausages.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00217-022-04063-2.

Non-sulfated steroidal glycosides cistoindosides from marine ‘old woman octopus’ Cistopus indicus attenuate pro-inflammatory lipoxygenase

Two non-sulfated steroidal glycosides, cistoindosides A-B were isolated from organic extract of the marine ‘old woman octopus’ Cistopus indicus (family Octopodidae). Their structures were characterized as 3β-acteoxy-23β-hydroxy-cholesta-9-ene-β-D-xylopyranoside (cistoindoside A) and 22,23-epoxy-3β-hydroxy-cholesta-5-ene-β-D-4'-O-acetoxy-xylopyranoside (cistoindoside B). Cistoindoside B, glycosylated with β-D-4'-O-acetoxy-xylopyranoside in conjunction with epoxy moieties displayed superior anti-inflammatory properties as acknowledged by its promising 5-lipoxygenase attenuation potential (IC50 2.11 µM) than the 5-lipoxygenase inhibitor drug zileuton (IC50 3.76 µM). The anti-inflammatory properties were corroborated by the promising antioxidant activities (IC50 ∼ 1.0–1.5 mM) of these steroid glycosides. Sizeably greater electronic properties, balanced hydrophobic-lipophilic properties (log POW ∼ 4.0) and comparatively lower steric factors were directly proportional to their bioactivities. Molecular simulation studies in the active sites of 5-lipoxygenase displaying lesser binding energies and inhibition constant (Ki) of cistoindoside B could be correlated with anti-inflammatory properties. Cistoindosides could be projected for their utilization as potential bioactive leads in functional food and pharmaceutical applications.

Exopolysaccharide from Lactobacillus rhamnosusZFM231 alleviates DSS‐induced colitis in mice by regulating gut microbiota

The exopolysaccharides (EPS) produced by Lactobacillus and other probiotics are associated with many benefits, such as immune regulation, antioxidant properties, antitumor effect, and regulation of intestinal microbe homeostasis. In the present study, the modulatory effect of EPS produced by Lactobacillus rhamnosus ZFM231 on the intestinal flora of mice with inflammatory bowel disease induced by dextran sulfate solution was investigated. Results indicated that weight loss, colonic length, the disease activity index score and colonic tissue damage in mice were significantly improved by EPS treatment. Compared with the model group, in the EPS-treated group, the diversity of and the composition of gut microbiota at both phylum and genus levels were found to recover to the levels of normal group, indicating the effective modulation on gut microbiota by EPS; short-chain fatty acids, including acetic acid, propionic acid and butyric acid produced by intestinal microbial metabolism, increased significantly; the level of anti-inflammatory factor transforning growth factor-β significantly increased and the level of pro-inflammatory factor tumor necrosis factor-α significantly decreased in the colonic cells of EPS-treated mice. It is clear that EPS produced by L. rhamnosus ZFM231 could find application in functional foods with the property of anti-ulcerative colitis. The experimental results provide new insights into the probiotic effect of EPS. © 2022 Society of Chemical Industry.

Optimization of ultrasonic-assisted extraction of flavonoids, polysaccharides, and eleutherosides from Acanthopanax senticosus using response surface methodology in development of health wine

Acanthopanax senticosus has been widely studied for its various active ingredients such as flavonoids, polysaccharides, and glycosides, while its application in the field of health food has not been reported. This study was conducted to optimize the ultrasonic-assisted extraction method of functional components in Acanthopanax senticosus based on single-factor experiments and Box-Behnken's response surface methodology. The optimal extraction conditions were determined as a solid-to-liquid ratio of 1:10 g/mL, extraction time of 35 min, and power of 200 W, and the actual yields of polysaccharides, flavonoids, and glycosides obtained were 37.27 ± 0.17, 14.83 ± 0.11, and 1.57 ± 0.03 mg/g, respectively. Then, the extract was added to rice wine to formulate Acanthopanax senticosus health wine and analyzed for its antioxidant activity, sensory characteristics, and flavor components. The results showed that ultrasonic-assisted extraction significantly improved the antioxidant activity and imparted good sensory characteristics to Acanthopanax senticosus health wine. HS-SPME-GC-MS analysis further demonstrated that the health wine has a unique flavor profile and can be further produced for application in functional foods and pharmaceuticals.

Structural Characteristics, Antioxidant and Hypoglycemic Activities of Polysaccharide from Siraitia grosvenorii.

The structural characterization, the in vitro antioxidant activity, and the hypoglycemic activity of a polysaccharide (SGP-1-1) isolated from Siraitia grosvenorii (SG) were studied in this paper. SGP-1-1, whose molecular weight is 19.037 kDa, consisted of Gal:Man:Glc in the molar ratio of 1:2.56:4.90. According to the results of methylation analysis, GC–MS, and NMR, HSQC was interpreted as a glucomannan with a backbone composed of 4)-β-D-Glcp-(1→4)-, α-D-Glcp-(1→4)-, and 4)-Manp-(1 residues. α-1,6 linked an α-D-Galp branch, and α-1,6 linked an α-D-Glcp branch. The study indirectly showed that SGP-1-1 has good in vitro hypoglycemic and antioxidant activities and that these activities may be related to the fact that the SGP-1-1’s monosaccharide composition (a higher proportion of Gal and Man) is the glycosidic-bond type (α- and β-glycosidic bonds). SGP-1-1 could be used as a potential antioxidant and hypoglycemic candidate for functional and nutritional food applications.

Whey protein-derived peptides: The impact of chicken pepsin hydrolysis upon whey proteins concentrate on their biological and technological properties

Whey proteins concentrate (WPC) was hydrolysed by chicken pepsin extract (CPE) compared with commercial porcine pepsin (PP), and the impact on biological (antioxidant) and technological properties (solubility, emulsifying activity index (EAI) and foaming activity (FA)) was studied. The results showed no significant differences between degree of hydrolysis (DH) values for both enzymes and ratios E/S tested. Total degradation of α-lactalbumin (α-La) was observed, while not for β-lactoglobulin (β-Lg). The hydrolysates produced by both enzymes had enhanced ABTS and ORAC antioxidant capacity. The hydrolysate obtained with both enzymes that had the maximum DH and antioxidant activity, also exhibited improvement of solubility at the pH range of 3–8. In contrast, EAI and FA of PP hydrolysates were enhanced at some pH points only. The obtained results with CPE are very promising and suggest that they could be suitable for application in functional food as natural ingredients.

Evaluation of Thymus pseudopulegioides plant extracts for total phenolic contents, antioxidant and antimicrobial properties

In this study, the ethanolic extracts of leaf and flower of Thymus pseudopulegioides Klokov & Des. Shost collected from Sultan Murat Plateau of Trabzon, Turkey was assessed for total phenolic contents and antioxidant activities. In addition, antibacterial activity of extracts against nine different bacterial strains (Staphylococcus aureus, Escherichia coli, Pseudomanas aeruginosa, Bacillus cereus, B. pumilus, B. subtilis, B. licheniformis, Listeria innocua and L. monocytogenes) were determined using the agar-dilution method. The extraction yields from leaf and flower were obtained as 10.95% and 0.94% (w/w), respectively. The total phenolic contents of leaf and flower extracts were determined as 7.84 and 0.39 mg GAE/g, respectively. Antioxidant activities of the ethanolic extracts from leaf and flower were evaluated by using DPPH radical scavenging, and leaf extract showed better antioxidant activity than flower extract. Although, minimum inhibitory concentration (MIC) of leaf extracts was determined as 1.028 mg/mL for all bacteria except B. subtilis (0.256 mg/mL), MIC of flower extracts for B. subtilis and B. pumilis was found as 0.256 mg/mL and for others was 0.512 mg/mL. The extracts of the tested parts of Thymus pseudopulegioides, especially the leaves might be valuable for functional food and therapeutic applications.

Inhibitory Effect of Fermented Flammulina velutipes Polysaccharides on Mice Intestinal Inflammation.

To investigate the effect of Flammulina velutipes polysaccharides (FVPs) on mice intestinal inflammation, FVPs were extracted from Flammulina velutipes (FV) using a solid anaerobic fermentation technique. The antioxidant and anti-inflammatory capacities of FVP and fermented FVP (FFVP) induced by lipopolysaccharide (LPS) were investigated in vitro and in vivo. The results showed that the yield of FFVP (9.44%) was higher than that of FVP (8.65%), but the molecular weight (MW) of FFVP (15,702 Da) was lower than that of FVP (15,961 Da). The antioxidant and anti-inflammatory capacities of FFVP were higher than that of FVP in preventing mice diarrhea, enhancing antioxidant capacities, and reducing the secretion and mRNA expression of interleukin-1β (IL-1β), IL-6, IL-18, and tumor necrosis factor-α (TNF-α). The anti-inflammatory mechanisms of FVP and FFVP were analyzed by inhibiting the activation of the NLRP3 signaling pathway using an LPS-induced mice model. This study indicated that FFVP could be used as a functional antioxidant, indicating a potential application in functional food and health products.

Anti-pseudo-allergic components in licorice extract inhibit mast cell degranulation and calcium influx.

Pseudo-allergic reactions (PARs) widely occur upon application of drugs or functional foods. Anti-pseudo-allergic ingredients from natural products have attracted much attention. This study aimed to investigate anti-pseudo-allergic compounds in licorice. The anti-pseudo-allergic effect of licorice extract was evaluated in rat basophilic leukemia 2H3 (RBL-2H3) cells. Anti-pseudo-allergic compounds were screened by using RBL-2H3 cell extraction and the effects of target components were verified further in RBL-2H3 cells, mouse peritoneal mast cells (MPMCs) and mice. Molecular docking and human MRGPRX2-expressing HEK293T cells (MRGPRX2-HEK293T cells) extraction were performed to determine the potential ligands of MAS-related G protein-coupled receptor-X2 (MRGPRX2), a pivotal target for PARs. Glycyrrhizic acid (GA) and licorice chalcone A (LA) were screened and shown to inhibit Compound48/80-induced degranulation and calcium influx in RBL-2H3 cells. GA and LA also inhibited degranulation in MPMCs and increase of histamine and TNF-α in mice. LA could bind to MRGPRX2, as determined by molecular docking and MRGPRX2-HEK293T cell extraction. Our study provides a strong rationale for using GA and LA as novel treatment options for PARs. LA is a potential ligand of MRGPRX2.

The Effect of Glycosylated Soy Protein Isolate on the Stability of Lutein and Their Interaction Characteristics.

Lutein is a natural fat-soluble carotenoid with various physiological functions. However, its poor water solubility and stability restrict its application in functional foods. The present study sought to analyze the stability and interaction mechanism of the complex glycosylated soy protein isolate (SPI) prepared using SPI and inulin-type fructans and lutein. The results showed that glycosylation reduced the fluorescence intensity and surface hydrophobicity of SPI but improved the emulsification process and solubility. Fluorescence intensity and ultraviolet–visible (UV–Vis) absorption spectroscopy results showed that the fluorescence quenching of the glycosylated soybean protein isolate by lutein was static. Through thermodynamic parameter analysis, it was found that lutein and glycosylated SPI were bound spontaneously through hydrophobic interaction, and the binding stoichiometry was 1:1. The X-ray diffraction analysis results showed that lutein existed in the glycosylated soybean protein isolate in an amorphous form. The Fourier transform infrared spectroscopy analysis results revealed that lutein had no effect on the secondary structure of glycosylated soy protein isolate. Meanwhile, the combination of lutein and glycosylated SPI improved the water solubility of lutein and the stability of light and heat.

GC–MS-Based Metabolites Profiling, In Vitro Antioxidant, Anticancer, and Antimicrobial Properties of Different Solvent Extracts from the Botanical Parts of Micromeria fruticosa (Lamiaceae)

The present study assessed the metabolites and bioactivities of Micromeria fruticosa plant parts extracted with various solvents (ethanol, n-hexane, and water) through the steeping extraction method. Thereafter, the extracts were analyzed using GC-MS. Moreover, the extracts were tested for their antioxidant, antimicrobial, and antitumor activities. The quali-quantitative analysis of Micromeria fruticosa crude extracts revealed the occurrence of 27 secondary metabolites. Some major bioactives identified were menthone, oleamide, pulegone, and menthol. Numerous antioxidant minerals, viz., Fe, Zn, and Mn, were present. The water extract of leaves exhibited the highest DPPH scavenging activity (89.73%), followed by the water extract of flowers (80.07%) at 100 μg/mL. The stems’ water extract demonstrated greater antimicrobial activity against all the bacteria species tested. The ethanolic leaf and aqueous stem extracts exhibited strong activity against C. albicans and E. coli. Flowers’ aqueous extract demonstrated the highest cytostatic effect on the colon cell line by reducing viability, followed by the leaves’ ethanol extract. The extraction solvents influenced the recovery of phytocompounds, and the highest pharmacological activities of the different extracts could be correlated to the presence of additional bioactives. Our results suggest that the Micromeria fruticosa plant is a favorable source of natural products with promising properties for potential nutraceutical and functional food applications.

Chicken Muscle-Derived ACE2 Upregulating Peptide VVHPKESF Inhibits Angiotensin II-Stimulated Inflammation in Vascular Smooth Muscle Cells via the ACE2/Ang (1–7)/MasR Axis

This study aimed to evaluate the modulatory effects of four chicken muscle-derived peptides [VRP, LKY, VRY, and VVHPKESF (V-F)] on angiotensin II (Ang II)-induced inflammation in rat vascular smooth muscle A7r5 cells. Only V-F could significantly attenuate Ang II-stimulated inflammation via the inhibition of NF-κB and p38 MAPK signaling, being dependent on the Mas receptor (MasR) not on the Ang II type 1 or type 2 receptor (AT1R or AT2R). V-F accelerated Ang II degradation by enhancing cellular ACE2 activity, which was due to ACE2 upregulation other than a direct ACE2 activation. These findings demonstrated that V-F ameliorated Ang II-induced inflammation in A7r5 cells via the ACE2/Ang (1-7)/MasR axis. Three peptide metabolites of V-F─VHPKESF, PKESF, and SF─were identified but were not considered major contributors to V-F's bioactivity. The regulation of peptide V-F on vascular inflammation supported its functional food or nutraceutical application in the prevention and treatment of hypertension and cardiovascular diseases.

Seaweeds as a Source of Functional Proteins

Protein is one of the major macronutrients essential in human nutrition. Protein sources especially animal sourced proteins are expensive, thus much work has been carried out to explore alternative protein sources. Seaweeds, or macroalgae, are emerging as one of the alternative protein sources. They are rich in protein with an excellent amino acid profile comparable to the other conventional protein sources. Seaweed protein contains bioactive components, such as free amino acids, peptides, lectins, and phycobiliproteins, including phycoerythrin and phycocyanin, among others. Seaweed proteins have been proved for their antihypertensive, antidiabetic, antioxidant, anti-inflammatory, antitumoral, antiviral, antimicrobial, and many other beneficial functional properties. Therefore, seaweed proteins can be a natural alternative source for functional food development. This paper discusses the compositional and nutritional aspects of seaweed protein, protein extraction techniques, functional properties of various seaweed proteins, as well as their safety for new product development and functional food applications.

Influence of different pretreatments and drying methods on the chemical compositions and bioactivities of Smilacis Glabrae Rhizoma.

BackgroundThe processing of medicinal plant materials is one of the important factors influencing the components and biological activities of TCMs. Smilax glabra Roxb. is an herbal vine widely distributed in China, and its dried rhizome (Smilacis Glabrae Rhizoma, SGR) is often used in traditional medicines and functional foods. The processing methods of fresh cutting for SGR slices have been included in ancient Chinese herbal works, some local standards of TCMs, and the current Chinese Pharmacopoeia. Nevertheless, to date, the scientific basis for the processing of fresh medicinal materials for SGR slices has not been revealed.MethodsTo optimize the processing method for preparing SGR slices from the fresh rhizomes, the chemical compositions of the un-pretreated and pretreated (boiling, steaming) samples before and after drying (sun-drying, shade-drying, oven-drying), and the contents of astilbin isomers in dried SGR were analyzed by UHPLC-Q-TOF-MS/MS and UHPLC-DAD methods, respectively. Then, the antioxidant, anti-inflammatory, xanthine oxidase and α-glucosidase inhibitory activities of the prepared SGR slices were investigated by biological assays.ResultsA total of fifty-two compounds were identified from the un-pretreated and pretreated samples and a total of forty-nine compounds were identified from the subsequently dried samples. After pretreated by boiling and steaming, the contents of neoastilbin, neoisoastilbin, and isoastilbin in the prepared samples all increased. As a quality marker of SGR, the content of astilbin was unchanged or decreased slightly compared with that in the un-pretreated samples. During the drying process, the contents of the four astilbin stereoisomers in the un-pretreated samples increased significantly, while those in the pretreated samples had a slight increase or decrease. The effects of different processing methods were sorted according to the bioactivities of the prepared SGR. As a result, SGR slices prepared with no pretreatment followed by a sun-drying process have a higher astilbin content, better bioactivities and more energy savings, representing the optimum processing method for SGR slices.ConclusionsThis study reveals the scientific basis for the processing of fresh medicinal materials for SGR slices. The results provide scientific information for the quality control of SGR and its rational applications in herbal medicines and functional foods.Graphical abstract Supplementary InformationThe online version contains supplementary material available at 10.1186/s13020-022-00614-7.

The functionality characteristic of the different formulas of nano-emulsion containing temulawak-red palm oil extract

Temulawak or Curcuma xanthorrhiza Roxb is usually used as traditional medicine (herbal medicine) by the community. Temulawak has antioxidant, anticancer, antihyperglycemic, anti-inflammatory, hepatoprotective and gastroprotective properties. Its application in functional foods is often limited by its low solubility and bioavailability in aqueous media. Nanoemulsion was made to improve the stability and bioavailability of Temulawak. Preparation of nano-emulsion was used high-pressure homogenization method. Temulawak extract in 10% red oil combined with Tween 20 and Tween 80 at a pressure of 1500 psi follows seven homogenization cycles. This method could produce nano-emulsion with the particle droplet below 150 nm. The antioxidant activity of temulawak extract nano-emulsion was 567.69 - 1324.62 ppm (AAE). The nano-emulsion preparation of temulawak extract was stable at 40°C storage, and its solubility increased (>90%). The in vitro bioaccessibility of temulawak extract nano-emulsion ranges from 23 to 98%. The optimum formula was produced with the concentration of 5% temulawak extract, 1% Tween 80, 2% Tween 20, with bioaccessibility of 98%. The optimum formula was tested in vivo bioavailability on white male rats with Spraque Dawley strain. The results showed that the nano-emulsion treatment had experienced faster elimination or metabolism than the temulawak extract. These results have important implications for effective delivery systems for bioactive compounds on temulawak.

Selenium accumulation and biotransformation in Streptococcus, Lactococcus, and Enterococcus strains

• Strains from LAB can accumulate and biotransform sodium selenite from growth medium. • First description of Se produced by Lactococcus lactis subsp. cremoris. • LAB transforms Se into selenocysteine and nanoparticles. • Se-enriched LAB can be used to develop new functional foods for humans. In the present study, we evaluated the ability of inorganic Se to accumulate and biotransform into different selenium species and Se nanoparticles (SeNPs) in seven bacteria from the Lactococcus , Streptococcus, and Enterococcus genera . The cytotoxicity of selenized strains to human adenocarcinoma Caco-2 and HT29 cells, the effect of selenium enrichment on antioxidant activity, and cell surface hydrophobicity of the tested strains were determined. When compared to that of lactococci and enterococci, increasing concentrations of sodium selenite in the cultivation media inhibited the growth of Streptococcus thermophilus strains. Thirty milligrams per liter Na 2 SeO 3 and its higher concentrations (50 and 100 mg/L) significantly reduced the viability of Caco-2 and HT29 cells by 40–75%. Bacteria mainly produced SeNPs and selenocysteine in Se form. Our results indicate that the tested strains can accumulate and transform inorganic selenium, which makes them promising candidates for selenization and possible application in functional foods.

Immunoregulatory activity of a low-molecular-weight heteropolysaccharide from Ganoderma leucocontextum fruiting bodies in vitro and in vivo.

The chemical structure of GLP-1, a novel water-soluble heteropolysaccharide purified Ganoderma leucocontextum fruiting bodies, has been characterized in our previous study. This study aimed to investigate the immunostimulatory activity of GLP-1 in vitro and in vivo by using RAW264.7 macrophages and cyclophosphamide-induced immunosuppressed mice model. Results showed that GLP-1 was able to enhance phagocytic activity and promote the production of reactive oxygen species, nitric oxide, tumor necrosis factor-α, interleukin-6, and monocyte chemoattractant protein-1 in RAW264.7 macrophages. Moreover, GLP-1 could activate mitogen-activated protein kinase, phosphatidylinositol-3-kinase/protein kinase B, and nuclear factor-kappa B signaling pathways through toll-like receptor 2 and dectin-1 receptors. Furthermore, GLP-1 increased the thymus index, serum immunoglobulin levels, and percentage of CD3+ T lymphocytes in cyclophosphamide-induced immunosuppressed mice. These results demonstrated that GLP-1 possessed significant immunostimulatory effects in vivo and in vitro and could be developed as an effective immunomodulator for application in functional foods.