DkDTX1/MATE1 mediates the accumulation of proanthocyanidin and affects astringency in persimmon.

Proanthocyanidins (PAs) is a kind of polyphenols widely distributed in plants, and their astringent properties can protect plants from herbivores and regulate fruit taste. There is a great difference in PA composition between astringent (A)-type and nonastringent (NA)-type persimmon. Here, we studied the potential of DkDTX1/MATE1 in regulating PAs composition through its preferred transport in persimmon fruit. The results of fluorescence microscope showed that the DkDTX1/MATE1 green fluorescence overlapped with the blue light emitted by PA. Overexpression of DkDTX1/MATE1 in persimmon leaves not only significantly increase the concentrations of PA, but also upregulated the expression of PA biosynthesis pathway genes. Further overexpression of DkDTX1/MATE1 in persimmon fruit discs and stable genetic transformation of DkDTX1/MATE1 also led to PA concentrations increased. Molecular docking and transporter assays showed that DkDTX1/MATE1 preferentially transported catechin, epicatechin gallate and epigallocatechin gallate. DkDTX1/MATE1 mainly bound to the PA precursors via serine at position 68. Our findings indicate that DkDTX1/MATE1 play a role in the accumulation of PAs in early stage of fruit development and affects the astringency of persimmon through preferential transport PA precursors, which provided a theoretical basis for the future use of metabolic engineering to regulate the composition of PAs in persimmon.

Three-dimensional fruit growth analysis clarifies developmental mechanisms underlying complex shape diversity in persimmon fruit.

The determination of fruit size and shape are of considerable interest in horticulture and developmental biology. Fruit typically exhibits three-dimensional structures characterized by geometric features that are dependent on the genotype. Although minor developmental variations have been recognized, few studies have fully visualized and measured these variations throughout fruit growth. Here, a high-resolution 3D scanner was used to investigate the fruit development of 51 persimmon (Diospyros kaki) cultivars with various complex shapes. We obtained 2380 3D models that fully represented fruit appearance, and enabled precise and automated measurements of shape features throughout fruit development, including horizontal and vertical grooves, length-to-width ratio, and roundness. The 3D fruit model analysis identified key stages that determined the shape attributes at maturity. Typically, genetic diversity was found in vertical groove development, and these grooves could be filled by tissue expansion in the carpel fusion zone during fruit development. In addition, transcriptome analysis of fruit tissues from groove and non-groove tissues revealed gene co-expression networks that were highly associated with groove depth variation. The presence of YABBY homologs was most closely associated with groove depth and indicated the possibility that this pathway is a key molecular contributor to vertical groove depth variation. Overall, our results revealed deterministic patterns of complex shape traits in persimmon fruit and showed that different growth patterns among tissues are the main factor contributing to the shape of both vertical and horizontal grooves.

Analysis of the Correlation between Persimmon Fruit-Sugar Components and Taste Traits from Germplasm Evaluation.

Persimmon fruits are brightly colored and nutritious and are fruits that contain large amounts of sugar, vitamins, mineral elements, and phenolic substances. The aim of this study was to explore the differences in fruit-sugar components of different persimmon germplasms and their relationships with phenotypic and flavor indices through the determination of phenotypes and sugar components and through electronic-tongue indices, which provided the basis and inspiration for the selection of different sugar-accumulating types of persimmon fruits and the selection of high-sugar persimmon varieties. Our results showed that persimmon germplasm fruit-sugar components were dominated by sucrose, glucose and fructose and that the remaining sugar components were more diverse but less distributed among the various germplasm types. Based on the proportion of each sugar component in the fruit, persimmon germplasms can be categorized into sucrose-accumulating and reduced-sugar-accumulation types. Sucrose-accumulating types are dominated by sucrose, galactose, fucose and inositol, while reduced-sugar-accumulation types are dominated by glucose, fructose, mannose-6-phosphate, and xylose. The content of sugar components in the germplasm persimmon of fruits of different types and maturity periods of also differed, with significant differences in sugar components between PCNA (pollination-constant non-astringent) and PCA (pollination-constant astringent) fruits. Cluster analysis classified 81 persimmon germplasms into three clusters, including cluster I-A, with low glucose and fructose content, and cluster I-B, with medium glucose, fructose, and sucrose contents. Cluster II was high in sucrose and fructose. Cluster III had high contents of glucose and fructose and low contents of sucrose and inositol.

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.

Density functional theory (DFT), molecular docking, and xanthine oxidase inhibitory studies of dinaphthodiospyrol S from Diospyros kaki L.

In this work, we investigated Diospyros kaki extract and an isolated compound for their potential as xanthine oxidase (XO) inhibitors, a target enzyme involved in inflammatory disorders. The prepared extract was subjected to column chromatography, and dinaphthodiospyrol S was isolated. Then XO inhibitory properties were assessed using a spectrophotometry microplate reader. DMSO was taken as a negative control, and allopurinol was used as a standard drug. The molecular docking study of the isolated compound to the XO active site was performed, followed by visualization and protein-ligand interaction. The defatted chloroform extract showed the highest inhibitory effect, followed by the chloroform extract and the isolated compound. The isolated compound exhibited significant inhibitory activity against XO with an IC50 value of 1.09 µM. Molecular docking studies showed that the compound strongly interacts with XO, forming hydrogen bond interactions with Arg149 and Cys113 and H-pi interactions with Cys116 and Leu147. The binding score of -7.678 kcal/mol further supported the potential of the isolated compound as an XO inhibitor. The quantum chemical procedures were used to study the electronic behavior of dinaphthodiospyrol S isolated from D. kaki. Frontier molecular orbital (FMO) analysis was performed to understand the distribution of electronic density, highest occupied molecular orbital HOMO, lowest unoccupied molecular orbital LUMO, and energy gaps. The values of HOMO, LUMO, and energy gap were found to be -6.39, -3.51 and 2.88 eV respectively. The FMO results indicated the intramolecular charge transfer. Moreover, reactivity descriptors were also determined to confirm the stability of the compound. The molecular electrostatic potential (MEP) investigation was done to analyze the electrophilic and nucleophilic sites within a molecule. The oxygen atoms in the compound exhibited negative potential, indicating that they are favorable sites for electrophilic attacks. The results indicate its potential as a therapeutic agent for related disorders. Further studies are needed to investigate this compound's in vivo efficacy and safety as a potential drug candidate.

Comparative Metabolomic and Transcriptomic Analyses Reveal Distinct Ascorbic Acid (AsA) Accumulation Patterns between PCA and PCNA Persimmon Developing Fruit.

Persimmon fruit has a high nutritional value and significantly varies between pollination-constant astringent (PCA) and pollination-constant non-astringent (PCNA) persimmons. The astringency type affects sugar, flavonoids, and tannin accumulation and is well known in persimmon fruit. However, the impact of the fruit astringency type on ascorbic acid (AsA) accumulation is limited. In this study, typical PCA varieties ('Huojing' and 'Zhongshi5') and PCNA varieties ('Yohou' and 'Jiro') of persimmon fruit were sampled at four developing stages (S1-S4) to provide valuable information on AsA content variation in PCA and PCNA persimmon. Persimmon fruit is rich in ascorbic acid; the AsA content of the four varieties 'Zhongshi5', 'Huojing', 'Jiro', and 'Youhou' mature fruit reached 104.49, 48.69, 69.69, and 47.48 mg/100 g. Fruit of the same astringency type persimmon showed a similar AsA accumulation pattern. AsA content was significantly higher in PCA than PCNA fruit at S1-S3. The initial KEGG analysis of metabolites showed that galactose metabolism is the major biosynthetic pathway of AsA in persimmon fruit. There were significant differences in galactose pathway-related metabolite content in developing PCA and PCNA fruit, such as Lactose, D-Tagatose, and D-Sorbitol content in PCA being higher than that of PCNA. Combined gene expression and WGCNA analyses showed that the expression of the GME (evm.TU.contig4144.37) gene was higher in PCA-type than in PCNA-type fruit in S1-S3 and exhibited the highest correlation with AsA content (r = 690 **, p < 0.01). Four hub genes, including the DNA methylation gene, methyltransferase gene, F-box, and Actin-like Protein, were identified as potential regulators of the GME gene. These results provide basic information on how astringency types affect AsA accumulation and will provide valuable information for further investigation on AsA content variation in persimmon fruit.

A Review: Pharmacological Effect of Natural Compounds in Diospyros kaki Leaves from the Perspective of Oxidative Stress.

Oxidative stress is caused by an imbalance between reactive oxygen species and antioxidant levels. Current research suggests that oxidative stress is one of the key factors in the development of many chronic diseases, and it has been a concern for many years. Many natural compounds have been studied for their special free-radical-scavenging properties. The major chemical constituents of the leaves of Diospyros kaki are flavonoids and triterpenoids, both of which are potential antioxidants that can prevent damage caused by reactive oxygen species or reactive nitrogen species and ameliorate diseases associated with oxidative stress. In addition to the major constituents such as flavonoids and triterpenoids, the leaves of Diospyros kaki include compounds such as phenylpropanoids, alkaloids, phenolic acids, and terpenes. Studies have shown these compounds have certain antioxidant and neuroprotective activities. Experiments have shown that flavonoids or the extracts from the leaves of Diospyros kaki have a variety of good pharmacological activities, which could activate oxidative stress and mitochondrial apoptosis, inhibit the proliferation of human prostate cancer cells and induce apoptosis. It also could achieve the effect of anti-cancer cell proliferation and induce apoptosis by regulating oxidative stress. The main chemical substance of the leaves of Diospyros kaki regulating oxidative stress may be these multi-hydroxyl structure compounds. These natural products exhibit significant antioxidant activity and are an important basis for the leaves of Diospyros kaki to treat human diseases by regulating oxidative stress. This review summarizes the structural types of natural products in the leaves of Diospyros kaki and elaborates the mechanism of the leaves of Diospyros kaki in neuroprotection, anti-diabetes, renal protection, retinal degenerative diseases, and anti-cancer from a new perspective of oxidative stress, including how it supplements other pharmacological effects. The chemical constituents and pharmacological effects of the leaves of Diospyros kaki are summarized in this paper. The relationship between the chemical components in the leaves of Diospyros kaki and their pharmacological effects is summarized from the perspective of oxidative stress. This review provides a reference for the study of natural anti-oxidative stress drugs.

Preferential transport activity of DkDTX5/MATE5 affects the formation of different astringency in persimmon.

Proanthocyanidins (PAs) are specialized metabolites that influence persimmon fruit quality. Normal astringent (A)-type and non-astringent (NA)-type mutants show significant variation in PA accumulation, but the influencing mechanism remains unclear. In this study, among the six identified DTXs/MATEs proteins associated with PA accumulation, we observed that allelic variation and preferential transport by DkDTX5/MATE5 induced variation in PA accumulation for A-type and NA-type fruit. The expression pattern of DkDTX5/MATE5 was correlated with PA accumulation in NA-type fruit. Upregulation and downregulation of DkDTX5/MATE5 promoted and inhibited PA accumulation, respectively, in the NA-type fruit. Interestingly, transporter assays of Xenopus laevis oocytes indicated that DkDTX5/MATE5 preferentially transported the PA precursors catechin, epicatechin, and epicatechin gallate, resulting in their increased ratios relative to the total PAs, which was the main source of variation in PA accumulation between the A-type and NA-type. The allele lacking Ser-84 in DkDTX5/MATE5 was identified as a dominantly expressed gene in the A-type and lost its transport function. Site-directed mutagenesis revealed that DkDTX5/MATE5 binds to PA precursors via Ser-84. These findings clarify the association between the transporter function of DkDTX5/MATE5 and PA variation, and can contribute to the breeding of new cultivars with improved fruit quality.

DkMYB14 is a bifunctional transcription factor that regulates the accumulation of proanthocyanidin in persimmon fruit.

Proanthocyanidins (PAs) are phenolic secondary metabolites that contribute to the protection of plant and human health. Persimmon (Diospyros kaki Thunb.) can accumulate abundant PAs in fruit, which cause a strong sensation of astringency. Proanthocyanidins can be classified into soluble and insoluble PAs; the former cause astringency but the latter do not. Soluble PAs can be converted into insoluble PAs upon interacting with acetaldehydes. We demonstrate here that DkMYB14, which regulates the accumulation of PA in persimmon fruit flesh, is a bifunctional transcription factor that acts as a repressor in PA biosynthesis but becomes an activator when involved in acetaldehyde biosynthesis. Interestingly, both functions contribute to the elimination of astringency by decreasing PA biosynthesis and promoting its insolubilization. We show that the amino acid Gly39 in the R2 domain and the ethylene response factor-associated amphiphilic repression-like motif in the C-terminal of DkMYB14 are essential for the regulation of both PA and acetaldehyde synthesis. The repressive function of DkMYB14 was lost after the mutation of either motif, and all activities of DkMYB14 were eliminated following the mutation of both motifs. Our results demonstrate that DkMYB14 functions as both a transcriptional activator and a repressor, directly repressing biosynthesis of PA and promoting its insolubilization, resulting in non-astringency in persimmon.

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).

Genome Sequence Resource of the Causal Agent of Persimmon Anthracnose, Colletotrichum horii Strain SD010 from China.

Colletotrichum horii is a main causal agent of persimmon (Diospyros kaki) anthracnose and is distributed widely in persimmon-producing areas of the world. Here, we report the first high-quality draft genome sequence of C. horii strain SD010. This will provide a reference for understanding adaptive evolution of genome structure, genes, and population diversity among members of the C. gloeosporioides species complex, and also help in understanding the mechanisms of host-pathogen interactions and improve management strategies of anthracnose.

Analysis of Pesticide Residues by QuEChERS Method and LC-MS/MS for a New Extrapolation of Maximum Residue Levels in Persimmon Minor Crop.

According to EU guidance SANCO/7525/VI/95 Rev. 10.3, residue data extrapolation from a surrogate major crop to a minor crop can be used for setting maximum residue levels (MRLs) with a reduced number of residue trials and representative selected pesticides. In this work, a QuEChERS method (citrate-buffered version and PSA with MgSO4 clean-up) and LC-ESI-MS/MS for the determination of boscalid, pyraclostrobin, fludioxonil, fluopyram and tebuconazole in persimmon was developed and validated according to EU Commission guidelines and afterwards used for the determination of residues in four field trials. Residue levels at harvest for each pesticide ranged between 0.347 and 0.028 mg/kg. After comparing EFSA residue data on apples, as the surrogate major crop, and conducting a consumer risk assessment, a proposal of residue data extrapolation to set MRLs in persimmons was performed. The results showed that pesticide residues in persimmons at harvest were consistently lower than residues in apples when substances were applied according to the same critical GAP. MRLs were set at 0.5 mg/kg for fludioxonil, 0.6 mg/kg for boscalid, 0.3 mg/kg for tebuconazole, 0.4 mg/kg for fluopyran and 0.3 mg/kg for pyraclostrobin. The ratio of the MRLs for apple/persimmon varied between 2.5 for boscalid and 1.25 for fluopyram, suggesting that residue extrapolation can be feasible, promoting the process of pesticide registration for minor crops and the settlement of MRL.

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.

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.

Phytohormone and integrated mRNA and miRNA transcriptome analyses and differentiation of male between hermaphroditic floral buds of andromonoecious Diospyros kaki Thunb.

BACKGROUND: Persimmon (Diospyros kaki Thunb.) has various labile sex types, and studying its sex differentiation can improve breeding efficiency. However, studies on sexual regulation patterns in persimmon have focused mainly on monoecy and dioecy, whereas little research has been published on andromonoecy. In order to reveal the sex differentiation regulation mechanism of andromonoecious persimmon, we performed histological and cytological observations, evaluated OGI and MeGI expression and conducted phytohormones assays and mRNA and small RNA transcriptome analyses of the male and hermaphroditic floral buds of the andromonoecious persimmon 'Longyanyeshi 1'. RESULTS: Stages 2 and 4 were identified as the critical morphological periods for sex differentiation of 'Longyanyeshi 1' by histological and cytological observation. At both stages, OGI was differentially expressed in male and hermaphroditic buds, but MeGI was not. This was different from their expressions in dioecious and monoecious persimmons. Meantime, the results of phytohormones assays showed that high IAA, ABA, GA3, and JA levels at stage 2 may have promoted male floral bud differentiation. However, high JA levels at stage 4 and high ZT levels at stages 2 and 4 may have promoted hermaphroditic floral bud differentiation. In these phytohormone biosynthesis and signaling pathways, 52 and 54 differential expression genes (including Aux/IAA, ARFs, DELLA, AHP, A-ARR, B-ARR, CYP735A, CRE1, PP2C, JAZ, MYC2, COI1, CTR1, SIMKK, ACO, and MPK6) were identified, respectively. During the development of male floral buds, five metacaspases genes may have been involved in pistil abortion. In addition, MYB, FAR1, bHLH, WRKY, and MADS transcription factors might play important roles in persimmon floral bud sex differentiation. Noteworthy, miR169v_1, miR169e_3, miR319_1, and miR319 were predicted to contribute to phytohormone biosynthesis and signaling pathways and floral organogenesis and may also regulate floral bud sex differentiation. CONCLUSION: The present study revealed the differences in morphology and phytohormones content between male and hermaphroditic floral buds of 'Longyanyeshi 1' during the process of sex differentiation, and identified a subset of candidate genes and miRNAs putatively associated with its sex differentiation. These findings can provide a foundation for molecular regulatory mechanism researching on andromonoecious persimmon.

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).

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.

Method Development and Validation for Simultaneous Determination of Six Flavonoids in Rat Eyes after Oral Administration of Diospyros kaki Leaves Extract by UPLC-MS/MS.

A robust ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique was proven effective for simultaneous characterization of six flavonoids including quercetin-3-O-beta-galactoside (Q3GAL), quercetin-3-O-beta-glucoside (Q3GLU), quercetin-3-(2-galloylglucoside) (Q3GG), kaempferol-3-O-beta-galactoside (K3GAL), kaempferol-3-O-beta-glucoside (K3GLU), and kaempferol-3-(2-galloylglucoside) (K3GG) in rat eyes. By investigation of corresponding validation parameters (linearity, selectivity, precision, accuracy, matrix effect, extraction recovery, and stability), the method was verified to be within current acceptable criteria. Thereafter, the validated method enabled quantification of the six compounds successful in rat eyes after oral administration of ethanol extract Diospyros kaki (EEDK) at 0, 3, 15, 35, 60, 120 min.

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.

Effects of a High-Molecular-Weight Polysaccharides Isolated from Korean Persimmon on the Antioxidant, Anti-Inflammatory, and Antiwrinkle Activity.

Persimmon (Diospyros kaki), a familiar and widespread fruit worldwide, is known to exhibit several physiological effects because of the presence of pharmacologically active compounds called phytochemicals. However, its high-molecular-weight compounds, particularly polysaccharides, have not been extensively studied. In this study, D. kaki extract (DK) was fractionated into low- and high-molecular-weight fractions (DK-L and DK-H, respectively) through ethanol fractionation, and their effects on antioxidant, anti-inflammatory, and antiwrinkle activities were investigated by an in vitro system. DK-H contained significantly higher contents of neutral sugar, uronic acid, and polyphenols compared to DK and DK-L. Furthermore, DK-H exhibited significantly improved pharmacological activities, such as antioxidant, anti-inflammatory, and antiwrinkle properties, compared to those of DK and DK-L, demonstrating that DK-H may play an important role in mediating the beneficial effects of persimmon. Sugar composition analysis and molecular characterization indicated that DK-H consisted of a galacturonic acid (GalA)-rich polysaccharide with a molecular weight of >345 kDa that mainly comprised GalA and small amounts of neutral sugar and polyphenol residues. These results suggest that the bioactive fraction DK-H is likely to be a GalA-rich pectic polysaccharide containing a small number of polyphenol residues, which may be a novel candidate in the pharmaceutical and cosmeceutical industries.