The flowers of Sophora japonica L. (SJF) and Robinia pseudoacacia L. (RPF) are edible and similar in appearance. The chemical constituents of SJF and RPF were compared by UPLC-Q-TOF-MS/MS and HPLC analysis in this study. A total of 29 and 19 constituents were identified in SJF and RPF, respectively. Flavonoid glycosides were the main constituents found in both flowers. The main aglycon moieties found in SJF were quercetin, kaempferol and isorhamnetin, whereas acacetin and kaempferol were the main ones found in RPF. Additionally, the content of flavonoids in SJF was significantly higher than that in RPF, as determined by HPLC. Rutin was the most dominant flavonoid in SJF with a content range of 72.31~88.15 mg/g, followed by quercetin (13.05~20.30 mg/g). Kaempferol-di(rhamnoside)-hexoside was the most dominant flavonoid in RPF with a content range of 25.94~30.00 mg/g. The distinct flavonoid profiles indicated the chemical non-equivalence of SJF and RPF. Therefore, RPF should not be considered a direct substitute for SJF in herbal medicine without further pharmacological and clinical validation.

Quercetin (Que) is a common flavonol with diverse biological activities. Cyclodextrins (CD) were used to enhance the oral bioavailability of Que and the underlying mechanisms were explored anew. The complexation reactions between Que and β-CD, 2-hydroxypropyl-β-CD (HP-β-CD) and methyl-β-CD (M-β-CD) were characterized by phase-solubility analysis and UV-Vis spectroscopic titration. The results revealed that HP-β-CD and M-β-CD exhibited stronger complexation capacity than native β-CD. It was novelty discovered that the reversible complexation reaction effectively suppressed the recrystallization of supersaturated Que. The solid complexes of Que with HP-β-CD and M-β-CD were prepared by spray drying with a drug loading capacity of 4.67 ± 0.08%. The SEM, XRD and DSC analyses showed Que lost its crystal structure in the complexes, while FTIR revealed the interactions between Que and CDs. The complexes significantly improved the dissolution and solubility of Que. In addition to being dissolved, approximately 25% Que existed in nanoparticle form in simulated gastrointestinal fluids. M-β-CD effectively maintained the particles in nanoscale, whereas HP-β-CD failed. After oral administration, Que mainly exists in rat circulating blood as glucuronide metabolites. Pharmacokinetics revealed that, compared with free Que, M-β-CD and HP-β-CD complexes did not significantly increase the blood concentration of Que, but increased the AUC(0-t) of quercetin glucuronide metabolite (Que-Glu) by 7.52 times and 4.44 times, respectively. The primary novelty of this work is the finding that in addition to solubility improvement, the suppression of Que recrystallization through CD complexation is also an important mechanism for bioavailability enhancement.

This study identified the chemical composition of Cyclocarya paliurus (Batal.) Iljinskaja flavonoids (CPF) and evaluated the antioxidant activities of 40% ethanol-eluted (EE)-CPF and 70% EE-CPF. Then we investigated whether 40% EE-CPF with stronger antioxidant capacity and higher flavonoid content exerted alleviative effects on dextran sulfate sodium (DSS)-induced colitis in mice. Our resulted showed that 40% EE-CPF improved colitis symptoms and colon tissue damage, as evidenced by the increased body weight, enhanced colon length, reduced disease activity index (DAI) score and increased goblet cells. Moreover, CPF reduced oxidative stress by regulating superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) levels, and restored intestinal barrier through Toll-like receptor 4(TLR4)/myeloid differentiation factor 88 (MyD88) signaling pathway, thereby exhibiting preventive effects on colitis in mice induced by DSS. Additionally, CPF regulated the composition of gut microbiota, leading to the increased beneficial microbiota, e.g., Lactobacillus and decreased harmful microbiota, e.g., Mucispirillum. Meanwhile, CPF increased the levels of short-chain fatty acids in cecal contents of mice. More interestingly, through microbiota functional analysis and correlation analysis, we proposed that CPF may alleviate colitis via microbial metabolism involved glycolysis u2162 and pyruvate fermentation pathways. Collectively, these results established a theoretical reference for developing CPF as anti-inflammatory agents in colitis.

An HPLC method was developed for simultaneous analysis of anthocyanins, chlorogenic acids and flavonoids in Roselle calyces, i.e., delphinidin-3-O-sambubioside, cyanidin-3-O-sambubioside, 3-caffeoylquinic acid, 5-caffeoylquinic acid, 4-caffeoylquinic acid, rutin, myricetin, quercetin and kaempferol. The quantification method was validated using calibration curves and stability and recovery tests. The recoveries of the nine analytes in Roselle calyces were in the range of 90.6% to 116.0%. Extraction optimization showed that chlorogenic acids and anthocyanins in Roselle could be extracted with water, whereas flavonoids required an ethanol-water solution. Quantitative analysis of six Roselle samples of different origins showed that the total anthocyanin content ranged from 3.65 to 13.81 mg D3SE g-1, and the total content of chlorogenic acids ranged from 4.52 to 6.32 mg g-1, but the flavonoid contents were very low. Stability analysis revealed that Roselle anthocyanins were prone to decomposition under high temperature and alkaline conditions, while chlorogenic acids were more stable.

gamma-Cyclodextrin metal-organic frameworks (gamma-CD-MOFs) were prepared using gamma-CD and KOH as the raw materials. The gamma-CD-MOFs showed a cubic shape with a particle size in the range of 400-700 nm and a Langmuir surface area of 406.88 m2 g-1. The gamma-CD-MOFs were used as a carrier of vitexin through adsorption in methanol, and the loading capacity was 34.9 mg g-1. XRD showed that vitexin lost its crystal structure, and FTIR showed that vitexin had secondary interactions with the carrier. The structure of the gamma-CD-MOFs collapsed quickly in water, thereby rapidly releasing the loaded vitexin. Using the carrier, the water solubility of vitexin increased by about 18.3 times from 31.79 mu g mL-1 to 581.78 mu g mL-1 in simulated gastric fluid at 37 degrees C. Meanwhile, the presence of gamma-CD inhibited the re-crystallization of vitexin and maintained its supersaturated status through the formation of complexes. A pharmacokinetic study showed that the oral bioavailability of vitexin was enhanced by 1.99 times in rats through the gamma-CD-MOF carrier.

To improve the liposolubility and stability of astilbin, a series of astilbin fatty acid esters (ASE) with different carbon chain lengths were successfully synthesized through lipase-catalyzed transesterification. It was found that the substrate conversions were between 97.86 % and 99.29 % as determined by HPLC analysis. An isomerization reaction was found in the enzymatic transesterification reaction system, and the product was neoastilbin fatty acid ester. The pathway of this isomerization reaction was investigated and a high relative content of ASE (>90 %) was obtained by controlling the reaction time (24 h-48 h). Moreover, the underlying mechanism by which a high substrate molar ratio (acyl donor: astilbin/mM: mM) inhibited the hydrolysis of ASE was revealed. The products were purified using preparative thin-layer chromatography, and their structures were identified by mass spectrometry, H-1 NMR and C-13 NMR analysis. Subsequently, the liposolubility, temperature, and pH stability of ASE were explored. It was found that the liposolubility and temperature stability of ASE were higher than those of astilbin. Additionally, ASE exhibited pH-dependent stability, maintaining structural integrity in acidic conditions but undergoing significant hydrolysis, isomerization, and decomposition in alkaline conditions. This study may expand the application of astilbin in foods and functional foods.

Roselle extract (RE) is rich in anthocyanins and chlorogenic acids. This study investigated the health-promoting effects of RE on lipid metabolism, oxidative stress, glycometabolism, and gut microbiota in obese mice fed a high-fat diet (HFD). The obesity model was induced by feeding mice a HFD, with RE supplementation added to their drinking water at concentrations of 2 and 4 mg/mL for 12 weeks. RE significantly reduced body weight gain and fat accumulation compared to the control group, alleviated hepatic steatosis, and improved insulin sensitivity. Additionally, RE restored antioxidative enzyme activities such as SOD and GSH-PX while reducing MDA levels. Transcriptomic analysis of the liver was performed to evaluate gene expression related to lipid metabolism, particularly in the PPAR signaling pathway. Gut microbiota analysis showed that RE increased beneficial bacteria and reduced the Firmicutes-to-Bacteroidetes ratio, suggesting an improvement in gut dysbiosis caused by the HFD. RE enhanced lipid metabolism, reduced oxidative stress, and improved insulin sensitivity in obese mice, potentially through modulation of the PPAR signaling pathway and gut microbiota, suggesting its potential as a therapeutic candidate for obesity-related metabolic disorders.

Physalis peruviana L. (P. peruviana) is an edible medicinal plant rich in bioactive phenolics. This study aimed to establish a hairy root (HR) culture of P. peruviana as a potential source for the synthesis of natural compounds. HRs were induced in P. peruviana using different Agrobacterium rhizogenes strains (R1601, C58C1, A4, and K599). Notably, K599 did not induce HR formation, whereas R1601, C58C1, and A4 yielded transformation frequencies of 57.78, 65.14, and 72.31%, respectively. Secondary metabolite production and antioxidant capacity were further examined in HRs induced using C58C1, R1601, and A4. It was found that A. rhizogenes R1601 induced the greatest increase (44% compared to that observed in the non-transformed culture). The methanolic extract of HRs induced by A. rhizogenes R1601 exhibited strong antioxidant capacity, with IC50 values of 1.41 mg DE/mL and 2.33 mg DE/mL for 2,2-diphenyl-1-picrylhydrazyl and 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid), respectively. The HR culture showed higher production of phenolic compounds and higher antioxidant capacity than the non-transformed cultures. Ultra-performance liquid chromatography time-of-flight tandem mass spectrometry was used to identify eight alkaloids, phenolics, and glycoside compounds. A. rhizogenes R1601 is emerging as a possible strain for the mass production of HR and bioactive phenolic compounds in P. peruviana.

To address the poor physicochemical stability of whey protein isolate (WPI)-stabilized emulsions, a novel chitooligosaccharide-caffeic acid covalent complexes (COS-CA) was first synthesized using lipase-catalyzed acylation to co-stabilized an O/W emulsion with WPI. Structural analyses confirmed that caffeic acid (CA) was successfully grafted onto the primary hydroxyl group of chitooligosaccharide (COS) through an ester bond, with a maximum grafting degree of 9.84 mg/g. Moreover, the WPI + COS-CA (physical mixture) exhibited stronger antioxidant activity than WPI alone or the WPI + COS (physical mixture), with DPPH radical scavenging, Fe3+ reducing, and Fe2+ chelating capacities of 94.42 %, 3.13 mM, and 58.26 %, respectively. In addition, the emulsifying activity index and emulsifying stability index of WPI + COS-CA were the highest (36.02 m2/g and 232.95 min). Subsequently, the emulsions stabilized by WPI, WPI + COS, and WPI + COS-CA were prepared. It was found that the WPI + COS-CA emulsion not only had an initial apparent viscosity 3 and 3.3 times higher than that of the WPI and WPI + COS emulsions, respectively, but also exhibited higher G' and G″ moduli and interfacial adsorbed protein content. Additionally, the WPI + COS-CA emulsion showed the lowest creaming index during storage and under environmental stresses. Moreover, this system significantly inhibited oxidation, reducing malondialdehyde by over 45 % and protein carbonyls by over 27 % after storage. Mechanism analysis indicated that COS-CA improved the stability of the WPI-stabilized emulsion by reinforcing interfacial strength and enhancing the antioxidant activity of WPI. This work proposes an innovative strategy for enhancing the physicochemical stability of WPI emulsion, which expands its potential applications in the food industry.

BackgroundObesity is increasing seriously worldwide. Inhibiting pancreatic lipase (PL) is an important way to prevent obesity. Baicalein, a dietary flavone, shows various physiological activities and has anti-obesity potential. Therefore, the inhibitory activity of baicalein on PL was investigated in present study.ResultsBaicalein exhibited a significant inhibitory effect on PL with IC50 value of 68 mu g/mL. Inhibition kinetics indicated that baicalein was a mixed-type inhibitor of PL. Fluorescence titration showed that baicalein could induce fluorescence quenching of PL. The binding constant (logKa) and number of binding sites were calculated as 5.40 and 1.11, respectively. In addition, synchronous fluorescence analysis revealed that the quenching ratio of baicalein to tryptophan was greater than that of tyrosine. Circular dichroism (CD) spectrum showed that the binding of baicalein affected the secondary structure of the enzyme protein. Through molecular docking analysis, it confirmed that baicalein could interact with amino acid residues in lipase, thus reducing the enzyme catalytic activity. In vivo studies showed that oral administration of baicalein with a does of 50 mg/kg bwt significantly reduced fat absorption and increased its fecal excretion in rats.ConclusionBaicalein showed inhibitory activity on PL in both in vitro and in vivo studies. It may be a safe and effective dietary supplement for obesity prevention.

Eupatorium chinense L. is highly valued for its medicinal properties. However, this species has a low propagation rate, so that the demand for the resource cannot be met adequately. The present study investigated the effects of various hormone combinations on callus formation, shoot induction, proliferation, and root growth in E. chinense L. to develop a rapid tissue-culture propagation method. Callus induction was achieved from leaf explants of aseptic in vitro plantlets on Murashige and Skoog (MS) medium fortified with 2.0 mg/L 6-benzyladenine (BA), 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.2 mg/L α-naphthaleneacetic acid (NAA), with a response rate of 75.66

Methyl-beta-cyclodextrin (M-beta-CD) exhibited the strongest complexation ability with astilbin (Ast) among three beta-CD derivatives, i.e., M-beta-CD, 2-hydroxyethyl-beta-CD, and sulfobutylether-beta-CD. The complexation reaction was investigated by UV spectroscopy, phase solubility, and NMR analysis. In simulated intestinal fluid (SIF), M-beta-CD resulted in significant changes of Ast UV spectra through hindering the dissociation of 7-OH by complexation. Their formation constants (Ka) in water, simulated gastric fluid (SGF), and SIF were determined by the UV titration method with values of 6148.2 +/- 558.9, 6596.3 +/- 346.1, and 4602.2 +/- 158.4 M--(1) , respectively. The Job's plot showed that M-beta-CD and Ast formed 1:1 inclusion complexes. Thermodynamic results revealed that the complexation was unfavourable at high temperature with negative Delta H and Delta S. The solubility of Ast linearly rose from 0.35 +/- 0.03-3.57 +/- 0.10 mM with the increase of M-beta-CD concentration from 0 to 5.0 mM. The NMR analysis implied that the A ring of Ast was preferentially embedded in M-beta-CD cavity. The Ast/M-beta-CD complexes were prepared by co-grinding method and further characterized by SEM, XRD, DSC and FT-IR. These analysis revealed that Ast lost its crystal structure after complexation, and the secondary interactions between Ast and the M-beta-CD were found. The complexes significantly improved the dissolution and solubility of Ast (>43 mg/mL). Furthermore, M-beta-CD suppressed the crystallization of Ast, thereby maintaining its supersaturated status. Besides, the stability of Ast in SIF and SGF was also improved. Pharmacokinetic studies showed that Ast/M-beta-CD complexes significantly enhanced the oral bioavailability of Ast by 11.89 times with C-max of 642.28 ng/mL in rat plasma.

Taxifolin, a natural dihydroflavonol compound, possesses notable anti-inflammatory properties and regulatory effects on intestinal microbiota. In this study, gelatinized-retrograded corn starch (GCS) was utilized as a carrier for colonic delivery of taxifolin, and its therapeutic efficacy against dextran sulfate sodium (DSS)-induced colitis in mice were systematically investigated. Taxifolin can integrate into the helical structure of starch, and the formation of GCS-Taxifolin complexes (GCS-Tax) significantly delayed the release of taxifolin in vitro. After oral administration of GCS-Tax, fecal excretion of taxifolin increased from 0.42 % to 10.89 % within 24 h compared to free taxifolin. Moreover, GCS-Tax facilitated the production of short-chain fatty acid in mice and effectively alleviated DSS-induced colitis symptoms, including weight loss, bloody stools, and colonic tissue damage. Additionally, GCS-Tax significantly suppressed proinflammatory factors such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and lipopolysaccharide (LPS), while elevating anti-inflammatory interleukin-10 (IL-10) level in mice serum. Furthermore, it restored intestinal mucosal barrier function by upregulating the expression of Mucin 2, Occludin, and zonula occludens-1 (ZO-1), reducing Beclin 1 expression, and exhibited hepatoprotective effects by enhancing total antioxidant capacity (T-AOC), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities. High-throughput sequencing analysis revealed that GCS-Tax improved intestinal flora diversity, reducing inflammation-related Bacterium 1 and Staphylococcus, while promoting the abundance of beneficial bacteria like Lachnospiraceae.

Quercetin is the most abundant flavonoid found in human diet. In the present study, the effects of quercetin on postprandial hyperglycemia (PPHG) were investigated in two ways. First, the complexes of quercetin and starch were prepared and characterized. Quercetin can occupy the helix of starch and interact with it through secondary bonds. The digestibility of starch decreased with the increasing proportion of quercetin in the complexes. Second, the inhibitory activities of quercetin on α-glucosidase and α-amylase were investigated in both in vitro and in vivo. Quercetin showed very strong inhibitory activity on α-glucosidase, while its inhibitory activity on α-amylase was far weaker than acarbose. Quercetin is the non-competitive inhibitor of α-glucosidase. Fluorescence titration showed that quercetin and α-glucosidase had one binding site and the binding constant lgKa was 3.70. After oral administration of quercetin–starch complexes in rats, the maximum level of postprandial blood glucose was reduced and delayed. Besides, quercetin also improved the PPHG after administration of sucrose and maltose in rats. Hence, it is deduced that the regulation of quercetin on PPHG is the result of its combination effects on starch and digestive enzymes. The complexes of quercetin and starch may be useful for the development of low-glycemic food.

Inhibition of pancreatic lipase (PL) is a strategy to prevent obesity. The inhibitory effects of Flos Sophorae Immaturus (FSI) extract and its main flavonoid components, rutin and quercetin, on PL were investigated. The contents of rutin and quercetin in FSI extract were 44.10 ± 1.33 % and 6.07 ± 1.62 %, respectively. The IC50 values of FSI extract, rutin and quercetin on PL were 322, 258 and 71 μg/mL, respectively. Rutin and quercetin inhibited PL in a reversible and noncompetitive manner. The combination of rutin and quercetin exhibited synergistic inhibitory effects at low concentration. The binding of rutin/quercetin with PL caused the fluorescence quenching of protein. Fluorescence titration showed the binding affinity of quercetin with PL protein was stronger than that of rutin. Circular dichroism analysis showed the binding changed the secondary structure of PL with an increase in random coil and a decrease in α-Helix and β-Sheet. Molecular docking revealed that rutin and quercetin could interact with the amino acid residues around the catalytic site through multiple secondary interactions. In vivo studies showed that FSI extract can reduce fat absorption and promote fecal fat excretion through inhibition of PL activity, and the effects were mainly due to rutin and quercetin.

Plant cell suspension culture has many advantages such as short production cycle and independence of natural conditions, and therefore shows a good application prospect in the production of plant secondary metabolite. In this paper, the regulatory effects of hormones on the growth and RosA (rosmarinic acid) synthesis of the cultured O. vulgare cells were comprehensively investigated, meanwhile, the mechanisms of the regulation were explored using integrated multi-omics analysis. The hormone screening and optimization tests revealed that 6-BA and NAA showed a significant promotion effect on cell growth, but 2,4-D displayed an inhibitory effect on growth, while KT, TDZ, ZT and IBA had no significant effect. The preferred hormone combination for cell growth was 4.0 mg/L NAA and 0.5 mg/L 6-BA, under which cell biomass was up to 20.25 g/L. Nine hormones, including KT, ZT, TDZ, 2,4-D, IBA, GA3, ETH, MeJA, and Spm, inhibited the synthesis of RosA in the cells, while ABA exhibited a significant promotion on the synthesis. The yield of RosA reached 283.54 mg/L under ABA induction, which was 3.47 times higher than that of the control. The integrated transcriptome and metabolome analysis showed that the expression levels of TFs (transcriptional regulators) such as GT-3A, MYB61, and WRKY72A were significantly up-regulated under ABA treatment, which in turn significantly promoted the expression of key genes for RosA synthesis like 4-coumarate CoA-ligase, cytochrome P450 monooxygenase, hydroxyphenylpyruvate reductase, abscisic-aldehyde oxidase and xanthoxin dehydrogenase, ultimately leading to a significant increase in RosA synthesis in cells.

A lipophilic piceid lipoate (PIL) was synthesized by enzymatic method to enhance the antioxidant activity of piceid and improve its state in oil system. The highest substrate conversion of 93.71 % was obtained in γ-valerolactone using Novozym 435 as a catalyst, with a piceid/lipoic acid ratio of 1:15 (mM/mM), an enzyme dosage of 40 mg/mL, and 4 Å molecular sieves at 400 mg/mL. The lipophilicity and antioxidant activity of piceid and PIL were evaluated, which found the introduction of α-lipoic acid markedly increased the lipophilicity and antioxidant activity of piceid. The oxidation inhibition rate of PIL in bulk oil was nearly 2.0 and 1.9 times higher than piceid and butylated hydroxytoluene, indicating that PIL is a promising antioxidant for oil-based foods. The antioxidant micro-mechanisms of PIL were revealed by computational simulations, which found the lipoic acid moiety in PIL molecule enhanced its free radical scavenging capacity and improved its solubility, dispersion, and migration in bulk oil.

SummaryObesity is becoming a top global health problem. Inhibition of digestive enzymes is an effective approach in weight management due to the reduction of energy absorption. In this study, the inhibitory effects of luteolin on α‐glucosidase and pancreatic lipase were investigated in vitro and in vivo via enzyme kinetics, fluorescence quenching, molecular docking and rat experiments. The results showed that luteolin inhibited α‐glucosidase and pancreatic lipase in a mixed‐type and uncompetitive manner with IC50 values of 29.31 and 42.74 μg mL−1 respectively. The binding of luteolin with the two enzymes caused the quenching of intrinsic fluorescence of protein. Molecular docking simulation revealed that luteolin could be located in the active pocket of α‐glucosidase and interact with the key amino acid residues. Moreover, a hydrophobic interaction and a van der Waals force were formed between luteolin and the key amino acid residues of pancreatic lipase (His264 and Ser153). In vivo studies further confirmed that luteolin could significantly decrease the absorption of glucose and triacylglyceride in rats by inhibiting α‐glucosidase and pancreatic lipase.

Luteolin loaded zein nanoparticles (Lut-ZNP) were prepared by using sodium caseinate as an electrostatic stabilizer. The formulation of the nanoparticles was optimized. Lut-ZNP were spray-dried, and the physicochemical properties were characterized by SEM, XRD, FT-IR and DSC. Then, the bioavailability of luteolin in rats was determined. Under the formulation of luteolin, zein and sodium caseinate with mass ratio of 1:5:15, the particle size, ζ-potential, encapsulation efficiency and loading efficiency of Lut-ZNP were 171.8 nm, -49.05 mV, 85.85% and 3.15%, respectively. Luteolin existed in the nanoparticles with amorphous form. Lut-ZNP exhibited good redispersibility in water after drying. Compared with free luteolin, the solubility, stability and release of luteolin in Lut-ZNP were greatly improved. Besides, the fecal excretion of luteolin in rats was significantly reduced after oral administration of Lut-ZNP. In addition to native luteolin, its metabolites including sulfate, glucuronidate and methylated glucuronidate were found in rat plasma. Lut-ZNP significantly increased the plasma concentrations of luteolin and its metabolites, and the bioavailability of luteolin was enhanced by 2.92 times.