Pulp or Peel? Comparative Analysis of the Phytochemical Content and Selected Cosmetic-Related Properties of Annona cherimola L., Diospyros kaki Thumb., Cydonia oblonga Mill. and Fortunella margarita Swingle Pulp and Peel Extracts.

Fruit peels might be a valuable source of active ingredients for cosmetics, leading to more sustainable usage of plant by-products. The aim of the study was to evaluate the phytochemical content and selected biological properties of hydroglycolic extracts from peels and pulps of Annona cherimola, Diospyros kaki, Cydonia oblonga, and Fortunella margarita as potential cosmetic ingredients. Peel and pulp extracts were compared for their antiradical activity (using DPPH and ABTS radical scavenging assays), skin-lightening potential (tyrosinase inhibitory assay), sun protection factor (SPF), and cytotoxicity toward human fibroblast, keratinocyte, and melanoma cell lines. The total content of polyphenols and/or flavonoids was significantly higher in peel than in pulp extracts, and the composition of particular active compounds was also markedly different. The HPLC-MS fingerprinting revealed the presence of catechin, epicatechin and rutoside in the peel of D. kaki, whereas kaempferol glucoside and procyanidin A were present only in the pulp. In A. cherimola, catechin, epicatechin and rutoside were identified only in the peel of the fruit, whereas procyanidins were traced only in the pulp extracts. Quercetin and luteolinidin were found to be characteristic compounds of F. margarita peel extract. Naringenin and hesperidin were found only in the pulp of F. margarita. The most significant compositional variety between the peel and pulp extracts was observed for C. oblonga: Peel extracts contained a higher number of active components (e.g., vicenin-2, kaempferol rutinoside, or kaempferol galactoside) than pulp extract. The radical scavenging potential of peel extracts was higher than of the pulp extracts. D. kaki and F. margarita peel and pulp extracts inhibited mushroom and murine tyrosinases at comparable levels. The C. oblonga pulp extract was a more potent mushroom tyrosinase inhibitor than the peel extract. Peel extract of A. cherimola inhibited mushroom tyrosinase but activated the murine enzyme. F. margarita pulp and peel extracts showed the highest in vitro SPF. A. cherimola, D. kaki, and F. margarita extracts were not cytotoxic for fibroblasts and keratinocytes up to a concentration of 2% (v/v) and the peel extracts were cytotoxic for A375 melanoma cells. To summarize, peel extracts from all analyzed fruit showed comparable or better cosmetic-related properties than pulp extracts and might be considered multifunctional active ingredients of skin lightening, anti-aging, and protective cosmetics.

Ultrasound-assisted deep eutectic solvent extraction of bioactive compounds from persimmon calyx.

This study aimed to investigate the ultrasound-assisted extraction of bioactive compounds from persimmon (Diospyros kaki) calyx by deep eutectic solvents (DES) with different molar ratios. For this reason, the prepared DES extracts' total phenolic-flavonoid compounds and antioxidant activities (1,1-diphenyl-2-picrilhydrazyl radical scavenging activity [DPPH•], Cupric Reducing Antioxidant Capacity (CUPRAC), and ferric reducing antioxidant power [FRAP]) were investigated as a result of the experimental design and optimization study conducted for this purpose. A sonication time of 20 min was determined as the optimal condition. Under these conditions, a molar ratio of 1.9:1 (lactic acid:choline chloride) and a water ratio of 70% provided the highest phenolic/flavonoid compounds and antioxidative activity. Correlations among water ratio, molar ratio, and sonication time were determined using principal component analysis (PCA). In conditions where total flavonoid compound, FRAP, and DPPH• are high due to PCA, it can be concluded that the sonication time is at high level; on the contrary, the water and molar ratios are at low level. In conclusion, ultrasound-assisted extraction using DES proved effective in persimmon calyx. Therefore, it can be recommended to use these environmentally friendly green solvents as an alternative to organic solvents in preparing extracts in various fields. PRACTICAL APPLICATION: This study shows the effectiveness of the ultrasound-assisted green extraction method using persimmon calyx specified as waste. These findings are compelling in the food industry in terms of consumers being now aware of green technology and the discovery that calyx is a good source of bioactive compounds.

Kaempferol-3-O-(2″-O-galloyl-ß-D-glucopyranoside): a novel neuroprotective agent from Diospryros kaki against cerebral ischemia-induced brain injury.

Our previous study demonstrated neuroprotective and therapeutic effects of a standardized flavonoid extract from leaves of Diospyros kaki L.f. (DK) on middle cerebral artery occlusion-and-reperfusion (MCAO/R)-induced brain injury and its underlying mechanisms. This study aimed to clarify flavonoid components responsible for the effects of DK using in vitro and in vivo transient brain ischemic models. Organotypic hippocampal slice cultures (OHSCs) subjected to oxygen- and glucose-deprivation (OGD) were performed to evaluate in vitro neuroprotective activity of DK extract and nine isolated flavonoid components. MCAO/R mice were employed to elucidate in vivo neuroprotective effects of the flavonoid component that exhibited the most potent neuroprotective effect in OHSCs. DK extract and seven flavonoids [quercetin, isoquercetin, hyperoside, quercetin-3-O-(2″-O-galloyl-ß-D-galactopyranoside), kaempferol, astragalin, and kaempferol-3-O-(2″-O-galloyl-ß-D-glucopyranoside) compound (9)] attenuated OGD-induced neuronal cell damage and compound (9) possessed the most potent neuroprotective activity in OHSCs. The MCAO/R mice showed cerebral infarction, massive weight loss, characteristic neurological symptoms, and deterioration of neuronal cells in the brain. Compound (9) and a reference drugs, edaravone, significantly attenuated these physical and neurological impairments. Compound (9) mitigated the blood-brain barrier dysfunction and the change of glutathione and malondialdehyde content in the MCAO mouse brain. Edaravone suppressed the oxidative stress but did not significantly affect the blood-brain barrier permeability. The present results indicated that compound (9) is a flavonoid constituent of DK with a potent neuroprotective activity against transient ischemia-induced brain damage and this action, at least in part, via preservation of blood-brain barrier integrity and suppression of oxidative stress caused by ischemic insult.

Investigation into Polyphenol Profile and Biological Activities of Enriched Persimmon/Apple Smoothies during Storage.

Smoothies are becoming an increasingly popular product as a healthy alternative to snacks. The consumer expects from this product that, apart from its nutritional value, it will also be qualitatively stable during storage. Therefore, in this study, original smoothies obtained with persimmon fruit puree and apple juice (Dk/Md) enriched with Arbutus unedo fruits, Myrtus communis purple berry extract, Acca sellowiana, and Crocus sativus petal juice were evaluated for their polyphenol composition, antioxidant activity, and inhibition on targeted digestive enzymes, over six months of storage. The amount of polyphenols evaluated by UPLC-PDA analysis decreased in six months from 23.5% for both Dk/Md and enriched C. sativus smoothies to 42.5% for enriched A. sellowiana, with anthocyanins the most sensitive compounds (71.7-100% loss). Values of antioxidant assays generally strongly decreased during the first three months (up to ca. 60%) and to a lesser extent in the following three months (0.4-27%). In addition, inhibitory activity on α-amylase, α-glucosidase, and pancreatic lipase, especially on the last two enzymes, was negatively affected by time storage. The outcome of this study indicates that persimmon fruit is a good option for producing smoothies, and enrichment with other plant extracts can enhance the bioactive compound content and biological activities. It is recommended that appropriate storage strategies to preserve the properties of those smoothies should be developed.

Therapeutic effects of a standardized-flavonoid Diospyros kaki L.f. leaf extract on transient focal cerebral ischemia-induced brain injury in mice.

This study aimed to investigate the neuroprotective and therapeutic effects of Diospyros kaki L.f. leaves (DK) on transient focal cerebral ischemic injury and underlying mechanisms using a middle cerebral artery occlusion (MCAO) model of mice. The animals received the MCAO operation on day 0. The daily administrations of DK (50 and 100 mg/kg, p.o) and edaravone (6 mg/kg, i.v), a reference drug with radical scavenging activity, were started 7 days before (pre-treatment) or immediately after the MCAO operation (post-treatment) and continued during the experimental period. Histochemical, biochemical, and neurological changes and cognitive performance were evaluated. MCAO caused cerebral infarction and neuronal cell loss in the cortex, striatum, and hippocampus in a manner accompanied by spatial cognitive deficits. These neurological and cognitive impairments caused by MCAO were significantly attenuated by pre- and post-ischemic treatments with DK and edaravone, suggesting that DK, like edaravone, has therapeutic potential for cerebral ischemia-induced brain damage. DK and edaravone suppressed MCAO-induced changes in biomarkers for apoptosis (TUNEL-positive cell number and cleaved caspase-3 protein expression) and oxidative stress (glutathione and malondialdehyde contents) in the brain. Interestingly, DK, but not edaravone, mitigated an increase in blood-brain permeability and down-regulation of vascular endothelial growth factor protein expression caused by MCAO. Although the exact chemical constituents implicated in the effects of DK remain to be clarified, the present results indicate that DK exerts neuroprotective and therapeutic activity against transient focal cerebral ischemia-induced injury probably by suppressing oxidative stress, apoptotic process, and mechanisms impairing blood-brain barrier integrity in the brain.

In-vitro antioxidant, lipoxygenase inhibitory, and in-vivo muscle relaxant potential of the extract and constituent isolated from Diospyros kaki (Japanese Persimmon).

Diospyros kaki (Japanese persimmon) is cultivated specious of the Diospyros genus. D. kaki is a multi-medicinal application in the folk system for the cure of ischemic stroke, angina, atherosclerosis, muscle relaxation, internal hemorrhage, hypertension, high cough, and infectious disease. The main objective of this study was the isolated bioactive metabolites from chloroform fractions of D. kaki. The extract and fractions were then tested for various in-vitro (antioxidant and lipoxygenase) and in-vivo (muscle relaxant) activities. The repeated chromatographic separation of chloroform extract afforded compound 1. Compound 1, n-hexane, and chloroform fractions were evaluated for in vitro antioxidant, lipoxygenase inhibitory, and in vivo muscle relaxant potency. The chloroform extract has 79.54% interaction with DPPH at higher concentrations (100 µg/ml) while the compound exhibited a maximum effect of 95.09% at 100 µg/ml. Compound 1 exhibited significant lipoxygenase inhibitory activity with an IC50 value of 36.98 µM followed by a chloroform extract of 57.09 µM. Similarly, compound 1 and chloroform extract showed excellent muscle relaxant effects at a higher dose. From this investigation, it is concluded that extracts and pure compounds exhibited promising antioxidant, lipoxygenase inhibitory, and muscle relaxant activity. This study excellently rationalizes the traditional usage of D. kaki in curing various diseases. Furthermore, the docking results indicate, that the isolated compound fits well into the active site of the lipoxygenase, and makes strong interactions with the target protein.

The Effects of Persimmon (Diospyros kaki L.f.) Oligosaccharides on Features of the Metabolic Syndrome in Zebrafish.

Metabolic syndrome has become a global health care problem since it is rapidly increasing worldwide. The search for alternative natural supplements may have potential benefits for obesity and diabetes patients. Diospyros kaki fruit extract and its oligosaccharides, including gentiobiose, melibiose, and raffinose, were examined for their anti-insulin resistance and obesity-preventing effect in zebrafish larvae. The results show that D. kaki oligosaccharides improved insulin resistance and high-fat-diet-induced obesity in zebrafish larvae, evidenced by enhanced ß-cell recovery, decreased abdominal size, and reduced the lipid accumulation. The mechanism of the oligosaccharides, molecular docking, and enzyme activities of PTP1B were investigated. Three of the oligosaccharides had a binding interaction with the catalytic active sites of PTP1B, but did not show inhibitory effects in an enzyme assay. The catalytic residues of PTP1B were typically conserved and the cellular penetration of the cell membrane was necessary for the inhibitors. The results of the mechanism of action study indicate that D. kaki fruit extract and its oligosaccharides affected gene expression changes in inflammation- (TNF-α, IL-6, and IL-1ß), lipogenesis- (SREBF1 and FASN), and lipid-lowering (CPT1A)-related genes. Therefore, D. kaki fruit extract and its oligosaccharides may have a great potential for applications in metabolic syndrome drug development and dietary supplements.

Aloe vera gel coating delays softening and maintains quality of stored persimmon (Diospyros kaki Thunb.) Fruits.

The effect of Aloe vera (AV) gel coating was studied on antioxidant enzymes activities, oxidative stress, softening and associated quality attributes of persimmon fruits. The fruits were coated with 0 and 50% AV-gel coating and stored for 20 days at 20 ± 1 ºC. AV-gel coated fruits exhibited considerably less weight loss, hydrogen peroxide level, electrolyte leakage and malondialdehyde content. AV-gel coated fruits had significantly higher ascorbate peroxidase, peroxidase, superoxide dismutase and catalase activities. In addition, AV-gel coating suppressed pectin methylesterase, polygalacturonase and cellulase activities and showed higher ascorbic acid, DPPH scavenging antioxidants and phenolics, and lower sugars and carotenoids. To the best of our knowledge, these results are the first evidence that AV-gel coating modulates the activities of cell wall degrading enzymes to delay ripening in climacteric fruits. So, AV-gel coating prohibited the onset of senescence by activating enzymatic antioxidant system, accumulating bioactive compounds and suppressing cell wall degradation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05412-5.

Effects of Different Chemicals on Sexual Regulation in Persimmon (Diospyros kaki Thunb.) Flowers.

Research on crop sexuality is important for establishing systems for germplasm innovation and cultivating improved varieties. In this study, androecious persimmon trees were treated with various concentrations of ethrel (100, 500, and 1,000 mg/L) and zeatin (1, 5, and 10 mg/L) to investigate the morphological, physiological, and molecular characteristics of persimmon. Ethrel at 1,000 mg/L and zeatin at 10 mg/L both significantly reduced the stamen length and pollen grain diameter in androecious trees. Ethrel treatment also led to reduced stamen development with degenerated cellular contents; zeatin treatment promoted the development of arrested pistils via maintaining relatively normal mitochondrial morphology. Both treatments altered carbohydrate, amino acid, and endogenous phytohormone contents, as well as genes associated with hormone production and floral organ development. Thereafter, we explored the combined effects of four chemicals, including ethrel and zeatin, as well as zebularine and 5-azacytidine, both of which are DNA methylation inhibitors, on androecious persimmon flower development. Morphological comparisons showed that stamen length, pollen viability, and pollen grain diameter were significantly inhibited after combined treatment. Large numbers of genes involving in carbohydrate metabolic, mitogen-activated protein kinase (MAPK) signaling, and ribosome pathways, and metabolites including uridine monophosphate (UMP) and cyclamic acid were identified in response to the treatment, indicating complex regulatory mechanisms. An association analysis of transcriptomic and metabolomic data indicated that ribosomal genes have distinct effects on UMP and cyclamic acid metabolites, explaining how male floral buds of androecious persimmon trees respond to these exogenous chemicals. These findings extend the knowledge concerning sexual differentiation in persimmon; they also provide a theoretical basis for molecular breeding, high-yield cultivation, and quality improvement in persimmon.

Persimmon Water Extract Suppresses Hepatic Lipotoxicity by Regulating Lipid Metabolites.

This study was performed to investigate the effects of persimmon (Diospyros kaki) on high-fat diet (HFD)-induced hepatic lipotoxicity. The compounds of persimmon water extract (PWE) were identified as gallic acid, glucogallin, 1-O-Galloyl-(2-O-acetyl)-glu, and trihydroxy-octadecadienoic acid. The PWE was ingested by C57BL/6 mice with an HFD for 8 weeks. The PWE improved glucose tolerance and suppressed weight gain by inhibiting increases in the weight of liver and adipose tissues. The results of serum biomarker analysis showed that PWE suppressed biomarkers such as liver injury and dyslipidemia. In ex vivo tests, reduction of oxidative stress and improvement of mitochondrial dysfunction were confirmed in the liver of PWE groups. In a molecular study, it was confirmed that PWE decreased lipid accumulation, insulin resistance, inflammation, and apoptosis in the liver. Finally, in a metabolite analysis of liver tissue using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), it was confirmed that PWE has an effect on lipid metabolism. In particular, PWE reduced phosphatidylcholines (PCs) and lysophosphatidylcholines (lysoPCs). Notably, it is presumed that the reduction of lysoPCs and PCs in the PWE group is related to the improvement of liver dysfunction due to lipotoxicity.

Nutritional and mineral composition of persimmon fruits (Diospyros kaki L.) from Central and Southern Italy.

In this study, the nutritional and mineral composition of Diospyros kaki fruits from Apulia, Campania, Lazio, Sardinia and Sicily regions was evaluated. Dietary fiber, mineral, pectin, polyphenol, and protein contents were evaluated. Particularly high are the contents of the dietary fiber and pectins. The mineral elements profile was interesting due to its modest content of sodium and high potassium concentration. Protein amounts were in line with the quantities for this fruit. The total polyphenol content of the fruits analysed was very variable, interesting for the quantities found both total and gallic acid. The results obtained confirmed the nutritional value of this fruit even for special dietary regimens such as hypertension and heart diseases and the genuineness of its cultivation in Central-Southern Italy.[Formula: see text].

Nephroprotective effect of persimmon leaves (Diospyros kaki L.f.) against CCl4-induced renal toxicity in Swiss Albino rats.

Diospyros kaki L.f. fruit and leaves are traditionally used for the treatment of hypertension, angina, internal hemorrhage, antithrombotic and anti-inflammatory effects.In the current study, the protective effects of ethyl acetate (Per-1), n-butanol (Per-2), and aqueous (Per-3) fractions of Diospyros kaki leaves against carbon tetrachloride (CCl4) induced nephrotoxicity in Swiss albino rats were tested. Animal were divided into nine groups; each group consists of six animals. The groups were : group I was untreated and kept as control, group II was treated with CCl4 only, group III (silymarin with CCl4); group IV (Per-1 100 mg/kg with CCl4);group V (Per-1 200 mg/kg with CCl4); group VI (Per-2 100 mg/kg with CCl4); group VII (Per-2 200 mg/kg with CCl4); group VIII (Per-3 100 mg/kg with CCl4); and group IX (Per-3 200 mg/kg with CCl4). Silymarin was used as standard drug. All tested fractions were found active (except Per-1 at low dose of 100 mg/kg) with significant value (p < 0.001) compared to CCl4 only group. Serum creatinine, malondialdehyde (MDA), and uric acid were significantly (p < 0.001) lowered in group VII-IX as compared to CCl4 only group. Similarly, total protein (TP) and non-protein sulfhydryls(NP-SH) level in kidney tissues were significantly (p < 0.001) elevated in the same groups compared to CCl4 only group. Further to check the cardio-protective potential, biochemical parameters such as LDH, creatine kinase, TP, MDA, and NP-SH levels in myocardial tissues were also estimated.These findings confirmed that the n-butanol and aqueous fractions are active and recommended for further bioactive phytoconstituents screening. Repeated column chromatography on silica gel G and sephadex-LH-20 of the active n-butanol fraction, four flavonoids were isolated. Based on the spectroscopic NMR data, compounds were identified as kaempferol (1), quercetin (2), astragalin (3), and rutin (4).

Rhamnogalacturonan I-rich polysaccharide isolated from fermented persimmon fruit increases macrophage-stimulatory activity by activating MAPK and NF-κB signaling.

BACKGROUND: Persimmon (Diospyros kaki) is a familiar and widespread fruit, cultivated worldwide. To date, physiological and chemical changes in fermented persimmon fruit and its active compounds have been rarely investigated. Moreover, comparative studies on the pharmacological activities of fermented persimmon fruit-derived compounds have not been reported. RESULTS: To investigate the effect of traditional fermented foods on immunostimulatory activity, non-fermented persimmon fruit (D. kaki, DK) and fermented persimmon fruit (fermented D. kaki, FDK) were prepared and further fractionated into low- and high-molecular weight fractions. FDK exhibited significantly higher activity toward the production of macrophage-stimulatory mediators compared with that of DK, and the high-molecular weight fraction (FDK-H) isolated from FDK was shown to have more potent activity than FDK. FDK-H not only increased the expression of immunostimulatory genes (TNF-α, IL-6, IL-12, and iNOS), but also stimulated the phosphorylation of both MAPK (ERK, JNK, and p38) and NF-κB (p65 and IκB) signaling molecules underlying macrophage activation. The putative chemical characteristic of FDK-H was identified as a pectic rhamnogalacturonan (RG) I-rich polysaccharide with a high molecular weight of 304 kDa containing galacturonic acid, arabinose, rhamnose, and galactose as the major monosaccharide units. CONCLUSION: The present study reveals that traditional fermentation is a useful method for increasing the macrophage-immunostimulatory activity of persimmon fruit, and the increased activity may be associated with structural modification of persimmon polysaccharides. This study may serve to identify a functional ingredient as an immunostimulatory agent, and our results may be applied to develop a new immunostimulatory product using FDK-H. © 2021 Society of Chemical Industry.

Chemical Constituents of the Leaves of Diospyros kaki (Persimmon).

Diospyros kaki (persimmon) leaves have long been utilized as traditional medicine for the treatment of ischemic stroke, angina, and hypertension and as a healthy beverage and cosmetic for anti-aging. This study aimed to isolate as many compounds as possible from an ethanol extract of the persimmon leaves to identify the biologically active compounds. The antioxidative effect of the ethyl acetate layer from the ethanol extract of the persimmon leaves was demonstrated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and online high-performance liquid chromatography-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (HPLC-ABTS) analysis. A new flavonoid, kaempferol-3-O-ß-d-2″-coumaroylgalactoside (1), and a new natural compound, kaempferol-3-O-ß-d-2″-feruloylglucoside (3) were isolated from the ethyl acetate layer, along with 25 previously known compounds, including fourteen flavonoids, one ionone, two coumarins, seven triterpenoids, and one acetophenone. Their structures were determined by the interpretation of spectrometric and spectroscopic data. All isolated compounds were rapidly evaluated using an online HPLC-ABTS assay, and of these, compounds 4-8, 11, 13, 15, and 16 clearly showed antioxidative effects. The amount of these compounds was 0.3-0.65% of the extract.

Immature Persimmon Suppresses Amyloid Beta (Aß) Mediated Cognitive Dysfunction via Tau Pathology in ICR Mice.

This study confirmed the ameliorating effect of immature persimmon (Diospyros kaki) ethanolic extract (IPEE) on neuronal cytotoxicity in amyloid beta (Aß)1-42-induced ICR mice. The administration of IPEE ameliorated the cognitive dysfunction in Aß1-42-induced mice by improving the spatial working memory, the short-term and long-term memory functions. IPEE protected the cerebral cholinergic system, such as the acetylcholine (ACh) level and acetylcholinesterase (AChE) activity, and antioxidant system, such as the superoxide dismutase (SOD), reduced glutathione (GSH) and malondialdehyde (MDA) contents. In addition, mitochondrial dysfunction against Aß1-42-induced toxicity was reduced by regulating the reactive oxygen species (ROS), mitochondrial membrane potential and ATP contents. In addition, IPEE regulated the expression levels of tau signaling, such as TNF-α, p-JNK, p-Akt, p-GSK3ß, p-tau, p-NF-κB, BAX and caspase 3. Finally, gallic acid, ellagic acid and quercetin 3-O-(6″-acetyl-glucoside) were identified as the physiological compounds of IPEE using ultra-performance liquid chromatography ion mobility separation quadrupole time-of-flight/tandem mass spectrometry (UPLC IMS Q-TOF/MS2).

Antidiarrheal, Antimicrobial, and Antioxidant Properties of the Aqueous Extract of Tunisian Persimmon (Diospyros kaki Thunb.) Fruits.

Medicinal plants were used to prevent and treat numerous gastrointestinal disorders owing, in part, to their antioxidant capacity. The protective effects of Diospyros kaki fruit aqueous extract (DKFAE) against castor oil (CO)-induced diarrhea was studied. The in vitro antioxidant and antibacterial properties were investigated using colorimetric and biochemical analyses. In vivo, 60 male rats were divided into 6 groups of 10 animals each (n = 10): control (C), CO, CO+various doses of DKFAE (100, 200, and 400 mg/kg, b.w., p.o.), and CO+loperamide (LOP, 10 mg/kg, b.w., p.o.).The DKFAE was rich in tannins and showed interesting antioxidant and antibacterial activities. The liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) analysis resulted in the identification of 16 phenolic compounds, among which quinic acid was the main one. The in vivo study showed that diarrhea was accompanied by an oxidative stress status as measured by an increase of lipid peroxidation, a decrease of glutathione and thiol group levels, as well as antioxidant enzyme activity depletion, such as glutathione peroxidase, superoxide dismutase, and catalase. The DKFAE administration significantly decreased the gastrointestinal transit in a dose-dependent manner. Besides, DKFAE protected against CO-induced diarrhea and intestinal fluid accumulation. Interestingly, DKFAE pretreatment counteracted all the oxidative stress status deregulation induced by CO intoxication. D. kaki fruit could be suggested for its strong protective effect against CO-induced acute diarrhea, which could be explained, in part, to its antimicrobial and antioxidant properties.

Pork Liver Pâté Enriched with Persimmon Coproducts: Effect of In Vitro Gastrointestinal Digestion on Its Fatty Acid and Polyphenol Profile Stability.

Agrofood coproducts are used to enrich meat products to reduce harmful compounds and contribute to fiber and polyphenol enrichment. Pork liver pâtés with added persimmon coproducts (3 and 6%; PR-3 and PR-6, respectively) were developed. Therefore, the aim was to study the effect of their in vitro gastrointestinal digestion on: the free and bound polyphenol profile (HPLC) and their colon-available index; the lipid oxidation (TBARs); and the stability of the fatty acid profile (GC). Furthermore, the effect of lipolysis was investigated using two pancreatins with different lipase activity. Forty-two polyphenols were detected in persimmon flour, which were revealed as a good source of bound polyphenols in pâtés, especially gallic acid (164.3 µg/g d.w. in PR-3 and 631.8 µg/g d.w. in PR-6). After gastrointestinal digestion, the colon-available index in enriched pâté ranged from 88.73 to 195.78%. The different lipase activity in the intestinal phase caused significant differences in bound polyphenols' stability, contributing to increased lipid oxidation. The fatty acids profile in pâté samples was stable, and surprisingly their PUFA content was raised. In conclusion, rich fatty foods, such as pâté, are excellent vehicles to preserve bound polyphenols, which can reach the colon intact and be metabolized by the intestinal microbiome.

Diospyros kaki extract protects myoblasts from oxidative stress-induced cytotoxicity via secretions derived from intestinal epithelium.

Under oxidative stress, reactive oxygen species (ROS) alter signal transduction and induce macromolecular damage in cells. Such oxidative damage can lead to sarcopenia, an age-related syndrome characterized by a progressive loss of mass and strength of skeletal muscles. Because food components do not directly come in contact with muscle cells, we focused on the effects of secretions produced by stimulated intestinal epithelial cells on oxidative stress in myoblast cells. An extract of Diospyros kaki was fractionated using different concentrations of ethanol. Each fraction showed different levels of antioxidant and phenolic compounds. The biological activity was evaluated using a Caco-2 cell coculture system. Secretions from Caco-2 cells exposed to 0.5 mg/mL D. kaki extract attenuated the oxidative stress-induced reduction of C2C12 cell viability, suggesting that the D. kaki extract could stimulate intestinal epithelial cells to produce secretions that reduce oxidative stress in myoblasts in vitro.

Identification of Phenolic Constituents and Inhibitory Activity of Persimmon Calyx and Shiteito against Tumor Cell Proliferation.

The persistent calyx on the fruit of Diospyros kaki, called "Shitei" in Japanese, is reported to contain phenolic compounds including condensed tannins. In this study, we isolated and characterized a new compound, together with 26 phenolic components, from the 70% acetone extract of Shitei, with structural elucidation based on spectroscopic analyses. In addition, we confirmed the presence of condensed tannins by 13C-NMR spectra, and the weight-average molecular weight was estimated by gel permeation chromatography (GPC) analysis. Next, Shiteito, a Kampo medicine consisting of Shitei, ginger, and clove clinically used to treat chronic hiccoughs occurring in association with anticancer drug treatments, and hot-water extracts of each of its components, were analyzed by HPLC, which determined that the main ingredient in Shiteito was derived from clove. We therefore isolated the ingredients and investigated their anti-tumor cell proliferative activity, together with Shiteito and Shitei extracts. As a result, Shiteito showed weak inhibition of hepatocellular carcinoma (Hep3B) cell proliferation at a high concentration. In contrast, ellagic acid, one of the main constituents of Shiteito, showed significant cytotoxicity against Hep3B cells, and significant inhibition of gastric adenocarcinoma (AGS) cell proliferation in a concentration-dependent manner. The ethyl acetate (EtOAc) fraction of the 70% acetone extract of Shitei significantly inhibited the proliferation of colon adenocarcinoma (Caco-2) and AGS cells at low to middle concentration, while showing strong cytotoxicity against Hep3B. These data indicate that Shiteito and Shitei extracts could enhance cancer drug treatment by preventing the associated chronic hiccups, and have the potential to be adjuvant treatments as well.

Prevention of the rise in plasma cholesterol and glucose levels by kaki-tannin and characterization of its bile acid binding capacity.

BACKGROUND: Bile acid-binding agents, such as cholestyramine and colesevelam, improve both cholesterol and glucose metabolism. Kaki-tannin, a polymerized condensed tannin derived from persimmon (Diospyros kaki), has been shown to have bile acid-binding capacity and a hypocholesterolemic effect. However, its effects on glucose metabolism have not been well studied, and the binding selectivity of kaki-tannin to bile acid molecules has not been reported. RESULTS: In vivo experiments using mice with high-fat diet-induced obesity showed that kaki-tannin intake (20 g kg-1 of the diet) increased fecal bile acid excretion by 2.3-fold and prevented a rise in plasma cholesterol levels and fasting plasma glucose levels. Kaki-tannin also suppressed the development of impaired glucose tolerance. To characterize the bile acid-binding capacity of kaki-tannin, we investigated its capacity to bind to eight types of bile acid and cholesterol in vitro. Kaki-tannin showed strong capacity to bind to lithocholic acid (85.5%), which has one hydroxy group. It also showed moderate capacity to bind to bile acids with two hydroxy groups (53.3%), followed by those with three hydroxy groups (39.0%), but kaki-tannin did not show binding capacity to cholesterol. These results suggest that the binding capacity of kaki-tannin to bile acids tends to decrease as the number of hydroxy groups increases. Interestingly, the binding capacity of kaki-tannin correlated with that of cholestyramine (correlation coefficient: r = 0.900). CONCLUSION: Our findings indicate that kaki-tannin binds preferentially to bile acids with fewer hydroxy groups and has beneficial effects on glucose metabolism as well as cholesterol metabolism. © 2020 Society of Chemical Industry.