Anti-Inflammatory Effects and Macrophage Activation Induced by Bioavailable Cinnamon Polyphenols in Mice.

SCOPE: Cinnamon is a commonly used spice and herb that is rich in polyphenols. Due to the limited bioavailability of oral polyphenols, it remains unclear to which extent they can reach cells and exert a biological effect. This study aims to investigate the impact of bioavailable cinnamon polyphenols on lipopolysaccharide (LPS)-stimulated macrophages. METHODS AND RESULTS: A polyphenol fraction is prepared from cinnamon (Cinnamomi ramulus) (CRPF) by boiling cinnamon in water and adsorbing the extract onto a hydrophobic resin. Mice are orally administered CRPF for 7 days and then subjected to three independent experiments: endotoxemia, serum collection, and macrophage isolation. Upon intraperitoneal lipopolysaccharide challenge, CRPF decreases serum levels of inflammatory cytokines, involving suppression of liver and spleen macrophages. When normal macrophages are cultured in serum obtained from CRPF-treated mice, they exhibit an anti-inflammatory phenotype. However, macrophages from CRPF-treated mice show an increased production of inflammatory cytokines when cultured in fetal bovine serum and stimulated with LPS. CONCLUSION: The study provides evidence for the presence of bioavailable cinnamon polyphenols with anti-inflammatory properties and macrophage activation. These findings suggest that cinnamon polyphenols have the potential to modulate macrophage function, which could have implications for reducing inflammation and improving immune function.

Evaluation of various clean-up sorbents in kale followed by LC-MS/MS analysis of pesticides.

Chlorophyll-rich samples, such as kale, interfere with the analysis of residual pesticides and adversely affect the integrity of tandem mass spectrometers. Dispersed solid-phase (d-SPE) extraction using graphitized carbon black effectively removes pigments from kale extracts; however, it also reduces the recoveries of 30 pesticides. To overcome this, alternative sorbents, including ENVI-Carb, ChloroFiltr, and Z-Sep+, were evaluated in this study. A sorbent combination based on 50 mg of Z-Sep+ was most advantageous (21/30), good precision (< 15%), excellent pigment removal capacity, and low matrix effect. The limit of quantification (0.0001-0.0040 mg/kg) was lower than the Korean maximum residue limits levels. The proposed method was validated according to international guidelines and applied to real kale samples. The results demonstrated that d-SPE using Z-Sep+ provides an effective strategy for ensuring mass spectrometry system integrity and improving the analytical accuracy in chlorophyll-rich samples. Supplementary information: The online version contains supplementary material available at 10.1007/s10068-022-01101-3.

Antioxidant capacity of 12 major soybean isoflavones and their bioavailability under simulated digestion and in human intestinal Caco-2 cells.

Soy isoflavones (SIs) show various health benefits, such as antioxidant and estrogenic effects. It is important to understand the bioaccessibility and bioavailability of SIs due to the close relation to their bioactivities. In this study, the antioxidant capacity, bioaccessibility, and bioavailability of 12 SIs were evaluated using radical-scavenging methods, simulations of human digestion, and Caco-2 cells in Transwell, respectively. All SIs were stable (91.1-99.2%) under gastric digestion conditions compared with the control (100%), whereas acetyl and malonyl conjugates were unstable (38.5% and 65.5%, respectively) under small intestinal digestion conditions. SI aglycones showed higher permeability (7-15 times) and cellular accumulation (8.8 times) than their glucosides. A small amount of SI conjugates was intact in the cell and in the basolateral side of each Transwell. These results suggest that SI conjugates, especially malonyl and acetyl forms, have incidental bioactivity after being metabolized to aglycones inside the cell.

Influence of production systems on phenolic characteristics and antioxidant capacity of highbush blueberry cultivars.

Blueberry is a crop grown worldwide due to the excellent quality and high polyphenol content of its fruit and tolerance to cold conditions. We investigated the influence of three production systems, namely an open field, heated greenhouse, and non-heated (plastic) greenhouse, on the phenolic characteristics (total phenolic, flavonoid, and anthocyanin content) and antioxidant capacities of "Spartan" (northern highbush), "Sharpblue" (southern highbush), and "O'Neal" (southern highbush) blueberry cultivars. The non-heated production system showed the highest phenolic characteristics and antioxidant capacity in "Spartan" and "O'Neal," while the open field production system showed the highest phenolic characteristics and antioxidant capacity in "Sharpblue." Derivatives of delphinidin and malvidin were two of the most abundant anthocyanins. The heated greenhouse production system resulted in larger amounts of delphinidin derivatives compared with the other production systems, while the blueberry grown in the non-heated greenhouse produced larger amount of malvidin derivatives. The anthocyanin profiles varied according to production system and blueberry cultivars. The principal component analysis loading plot of blueberries for individual anthocyanins explained over 95% of the total variance. In summary, the results of this study suggest that a strategic approach to blueberry production could elevate the phenolic content and antioxidant capacity of cultivated blueberry. PRACTICAL APPLICATION: The highbush blueberry, a rich source of bioactive polyphenols, is a popular fruit. The microclimate of the production system of highbush blueberries affects the concentrations of antioxidative phenolic compounds such as anthocyanins. Therefore, discovering and applying the appropriate method of production for each blueberry cultivar could facilitate production of high-quality blueberries rich in phenolic antioxidants.

Chemometric Analysis of Extracts and Fractions from Green, Oxidized, and Microbial Fermented Teas and Their Correlation to Potential Antioxidant and Anticancer Effects.

Previous reports on phytochemicals in green tea (GT) and processed teas mainly focused on more representative compounds such as catechins. Here, we focus on the insignificantly studied non-catechin components in tea extracts, and explore the multivariate correlation between diverse phenolic compounds in tea and the in vitro antioxidant and anticancer effects. Extracts from GT and four types of processed teas were further divided into hydrophilic and hydrophobic fractions, whose phenolic compositions and antioxidant capacities were quantified using HPLC-MS and three antioxidant assays, respectively. For three types of teas, the anticancer effects of their extracts and fractions were assessed using cancer cell lines. The hydrophobic fractions had lower antioxidant capacities than the corresponding hydrophilic fractions, but exhibited superior antiproliferative effects on cancer cells compared with the whole extract and the hydrophilic fraction. Partial least squares-discriminant analysis revealed a strong correlation between the anticancer effects and the theaflavins and flavonols. Therefore, in addition to catechins, the hydrophobic fraction of tea extracts may have beneficial health effects.

Anti-Inflammatory Effect of Flavonoids from Brugmansia arborea L. Flowers.

Brugmansia arborea L. (Solanaceae), commonly known as "angel's trumpet," is widely grown in North America, Africa, Australia, and Asia. It has been mainly used for ornamental purposes as well as analgesic, anti-rheumatic, vulnerary, decongestant, and anti-spasmodic materials. B. arborea is also reported to show anti-cholinergic activity, for which many alkaloids were reported to be principally responsible. However, to the best of our knowledge, a phytochemical study of B. arborea flowers has not yet been performed. Four flavonol glycosides (1-4) and one dihydroflavanol (5) were for the first time isolated from B. arborea flowers in this study. The flavonoids showed significant antioxidant capacities, suppressed nitric oxide production in lipopolysaccharide (LPS)-treated RAW 264.7 cells, and reduced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) protein production increased by LPS treatment. The contents of compounds 1-4 in n-BuOH fraction were determined to be 3.8 ± 0.9%, 2.2 ± 0.5%, 20.3 ± 1.1%, and 2.3 ± 0.4%, respectively, and that of compound 5 in EtOAc fraction was determined to be 12.7 ± 0.7%, by HPLC experiment. These results suggest that flavonol glycosides (1-4) and dihydroflavanol (5) can serve as index components of B. arborea flowers in standardizing anti-inflammatory materials.

Stability and Fermentability of Green Tea Flavonols in In-Vitro-Simulated Gastrointestinal Digestion and Human Fecal Fermentation.

Flavonols, the second most abundant flavonoids in green tea, exist mainly in the form of glycosides. Flavonols are known to have a variety of beneficial health effects; however, limited information is available on their fate in the digestive system. We investigated the digestive stability of flavonol aglycones and glycosides from green tea under simulated digestion and anaerobic human fecal fermentation. Green tea fractions rich in flavonol glycosides and aglycones, termed flavonol-glycoside-rich fraction (FLG) and flavonol-aglycone-rich fraction (FLA) hereafter, were obtained after treatment with cellulase and tannase, respectively. Kaempferol and its glycosides were found to be more stable in simulated gastric and intestinal fluids than the derivatives of quercetin and myricetin. Anaerobic human fecal fermentation with FLG and FLA increased the populations of Lactobacilli spp. and Bifidobacteria spp. and generated various organic acids, such as acetate, butyrate, propionate, and lactate, among which butyrate was produced in the highest amount. Our findings indicate that some stable polyphenols have higher bioaccessibilities in the gastrointestinal tract and that their health-modulating effects result from their interactions with microbes in the gut.

Antioxidative, Anti-Inflammatory, and Anticancer Effects of Purified Flavonol Glycosides and Aglycones in Green Tea.

(1) Background: Extensive research has focused on flavan-3-ols, but information about the bioactivities of green tea flavonols is limited. (2) Methods: In this study, we investigated the antioxidative, anti-inflammatory, and anticancer effects of flavonol glycosides and aglycones from green tea using in vitro cell models. The fractions rich in flavonol glycoside (FLG) and flavonol aglycone (FLA) were obtained from green tea extract after treatment with tannase and cellulase, respectively. (3) Results: FLG and FLA contained 16 and 13 derivatives, respectively, including apigenin, kaempferol, myricetin, and quercetin, determined by mass spectrometry. FLA exhibited higher radical-scavenging activity than that of FLG. FLG and FLA attenuated the levels of intracellular oxidative stress in neuron-like PC-12 cells. The treatment of RAW 264.7 murine macrophages with FLG and FLA significantly reduced the mRNA expression of inflammation-related genes in a dose-dependent manner. Furthermore, FLG and FLA treatments decreased the viability of the colon adenoma cell line DLD-1 and breast cancer cell line E0771. Moreover, the treatment with FLG or FLA combined with paclitaxel had synergistic anticancer effects on the DLD-1 cell line. (4) Conclusions: Flavonols from green tea exerted beneficial effects on health and may be superior to flavan-3-ols.

High-efficiency enzymatic production of α-isoquercitrin glucosides by amylosucrase from Deinococcus geothermalis.

Isoquercitrin (IQ, quercetin-3-O-ß-d-glucopyranoside) has diverse biological functions, such as anti-oxidant and anti-cancer activity, but its use is limited by poor solubility and bioavailability. Enzymatically modified IQ (EMIQ) is a mixture of transglycosylated IQs that have better solubility and bioavailability than do quercetin and IQ. Two different enzymes, cyclodextrin glucanotransferase (CGTase) and amylosucrase (ASase), have the transglycosylation activity to produce EMIQ. Both enzymes produce a variety of EMIQs including IQ, IQ-glucoside (IQ-G1), IQ-diglucoside (IQ-G2), and IQ-triglucoside (IQ-G3). ASase had a higher bioconversion yield from IQ to EMIQ (97.6%) than did CGTase (76.8%). In addition, the yield of IQ-G3, which was the most bioavailable form, was higher with ASase (46%) than with CGTases (8%). Taken together, these results suggest that ASase can be used to synthesize EMIQ in a simple and specific process.

Enzymatic synthesis of α-flavone glucoside via regioselective transglucosylation by amylosucrase from Deinococcus geothermalis.

α-Flavone glycosides have beneficial properties for applications in the pharmaceutical, cosmetic, and food industries. However, their chemical syntheses are often limited by a low efficiency or scarcity of substrates. In this study, α-flavone glucosides were enzymatically synthesized by amylosucrase from Deinococcus geothermalis (DGAS) using sucrose and various flavones as a donor for glucosyl units and acceptors, respectively. Luteolin was the most effective acceptor in the transglucosylation reaction using DGAS among nine flavone materials (apigenin, chrysin, 6,7-dihydroxyflavone, homoorientin, 7-hydroxyflavone, isorhoifolin, luteolin, luteolin-3',7-diglucoside, and orientin). The highest production yield of luteolin glucoside was 86%, with a 7:1 molar ratio of donor to acceptor molecules, in 50 mM Tris-HCl buffer (pH 7) at 37°C for 24 h using 2 U of DGAS. The synthesized luteolin glucoside was identified as luteolin-4'-O-α-D-glucopyranoside with a glucose molecule linked to the C-4' position on the B-ring of luteolin via an α-glucosidic bond, as determined by 1H and 13C nuclear magnetic resonance. This result clearly confirmed that the glucosylated luteolin was successfully synthesized by DGAS and it can be applied as a functional ingredient. Furthermore, this approach using DGAS has the potential to be utilized for the synthesis of various glucosylated products using different types of polyphenols to enhance their functionalities.

Effect of Orally Administered Atractylodes macrocephala Koidz Water Extract on Macrophage and T Cell Inflammatory Response in Mice.

The rhizome of Atractylodes macrocephala Koidz (AM) is a constituent of various Qi booster compound prescriptions. We evaluated inflammatory responses in macrophages and T cells isolated from mice following oral administration of AM water extract (AME). Peritoneal exudate cells were isolated from thioglycollate-injected mice and alterations in scavenger receptors were examined. Peritoneal macrophages were stimulated with lipopolysaccharide (LPS). Serum cytokine responses to intraperitoneal LPS injection were also evaluated. Splenocytes were isolated and their composition and functional responses were measured. The content of atractylenolide I and atractylenolide III, known anti-inflammatory ingredients, in AME was 0.0338 mg/g extract and 0.565 mg/g extract, respectively. AME increased the number of SRA(+)CD11b(+) cells in response to thioglycollate. Peritoneal macrophages isolated from the AME group showed no changes in inflammatory markers such as tumor necrosis factor- (TNF-) α, interleukin- (IL-) 6, inducible nitric oxide synthase, and cyclooxygenase-2 but exhibited a decrease in CD86 expression. Interestingly, AME decreased the serum levels of TNF-α and IL-6 upon intraperitoneal injection of LPS. Regarding the adaptive immune system, AME increased the CD4(+) T cell population and major histocompatibility complex class II molecule expression in the spleen, and cultured splenocytes from the AME group showed increased production of IL-4 concurrent with decreased interferon-γ production during T cell activation. AME promoted the replenishment of peritoneal macrophages during the inflammatory response but its anti-inflammatory activity did not appear to be mediated by the modulation of macrophage activity. AME also altered the immune status of CD4 T cells, promoting the Th2 response.