Autumn Olive (Elaeagnus umbellata Thunb.) Berries Improve Lipid Metabolism and Delay Aging in Middle-Aged Caenorhabditis elegans.

This study evaluated the positive effects of autumn olive berries (AOBs) extract on delaying aging by improving lipid metabolism in middle-aged Caenorhabditis elegans that had become obese due to a high-glucose (GLU) diet. The total phenolic content and DPPH radical scavenging abilities of freeze-dried AOBs (FAOBs) or spray-dried AOBs (SAOBs) were examined, and FAOBs exhibited better antioxidant activity. HPLC analysis confirmed that catechin is the main phenolic compound of AOBs; its content was 5.95 times higher in FAOBs than in SAOBs. Therefore, FAOBs were used in subsequent in vivo experiments. FAOBs inhibited lipid accumulation in both the young adult and middle-aged groups in a concentration-dependent manner under both normal and 2% GLU conditions. Additionally, FAOBs inhibited ROS accumulation in a concentration-dependent manner under normal and 2% GLU conditions in the middle-aged worms. In particular, FAOB also increased body bending and egg production in middle-aged worms. To confirm the intervention of genetic factors related to lipid metabolism from the effects of FAOB, body lipid accumulation was confirmed using worms deficient in the daf-16, atgl-1, aak-1, and akt-1 genes. Regarding the effect of FAOB on reducing lipid accumulation, the impact was nullified in daf-16-deficient worms under the 2% GLU condition, and nullified in both the daf-16- and atgl-1-deficient worms under fasting conditions. In conclusion, FAOB mediated daf-16 and atgl-1 to regulate lipogenesis and lipolysis in middle-aged worms. Our findings suggest that FAOB improves lipid metabolism in metabolically impaired middle-aged worms, contributing to its age-delaying effect.

Biomimetic facile synthesis of zinc oxide and copper oxide nanoparticles from Elaeagnus indica for enhanced photocatalytic activity.

The present study focused on synthesizing ZnO nanoparticles (NPs) and CuO NPs using Elaeagnus indica leaf extract as reducing and stabilizing agents using Zn(O2CCH3)2 and Cu2SO4, respectively, for the first time. We have confirmed the formation of aggregated ZnO NPs and CuO NPs with phytochemicals by various spectral analyses and electron microscopy studies. The size of synthesized ZnO NPs and CuO NPs were in the range of 20-30 nm and 30-40 nm, respectively. The antimicrobial activity of ZnO NPs at 75 µg concentration is superior against Salmonella typhimurium, Klebsiella pneumonia, Bacillus subtilis, Staphylococcus epidermidis, and Aspergillus niger. While CuO nanoparticles with 75 µg concentration effectively inhibited S. typhimurium, B. subtilis, S. epidermidis, and A. niger. Phytochemicals and reactive oxygen species generated by the prepared NPs may account for the antimicrobial effects observed. The photodegradation of methylene blue by ZnO NPs and CuO NPs was 91% and 76%, respectively, for 6 h of sunlight exposure. CuO NPs and ZnO NPs have different intrinsic properties and phytochemical compositions; hence ZnO NPs photodegrade faster than CuO NPs even though ZnO has higher bandgap energy than CuO. Consequently, CuO and ZnO NPs produced from E. indica leaf extract might be utilized as antimicrobials and photocatalysts in the future.

Involvement of NF-κB in the reversal of CYP3A down-regulation induced by sea buckthorn in BCG-induced rats.

Previous studies reported that sea buckthorn (Hippophae rhamnoides L., Elaeagnaceae, HRP) exhibits hepatoprotective effects via its anti-inflammatory and antioxidant properties as well as its inhibitory effects on collagen synthesis. However, it is unclear whether this hepatoprotective effect is also achieved by regulating liver drug metabolism enzyme pathways. Herein, we examined the regulatory effect of HRP on cytochrome P450 3A (CYP3A) in rats with immune liver injury, and explored the molecular mechanism of its hepatoprotective effect. Rat models of immunological liver injury were induced by intravenous injections of Bacillus Calmette-Guerin (BCG; 125 mg kg-1; 2 wks). Specific protein levels were detected by ELISA or western blot, and CYP3A mRNA expression was detected by RT-PCR. High-performance liquid chromatography (HPLC) detected relative changes in CYP3A metabolic activity based on the rates of 1-hydroxylation of the probe drug midazolam (MDZ). BCG pretreatment (125 mg kg-1) significantly down-regulated liver CYP3A protein expression compared with the control, metabolic activity, and transcription levels while up-regulating liver NF-κB, IL-1ß, TNF-α and iNOS. HRP intervention (ED50: 78 mg kg-1) moderately reversed NF-κB, inflammatory cytokines, and iNOS activation in a dose-dependent manner (P < 0.05), and suppressed CYP3A down-regulation (P < 0.05); thereby partially alleviating liver injury. During immune liver injury, HRP may reverse CYP3A down-regulation by inhibiting NF-κB signal transduction, and protect liver function, which involves regulation of enzymes transcriptionally, translationally and post-translationally. The discovery that NF-κB is a molecular target of HRP may initiate the development and optimization of a clinical therapeutic approach to mitigate hepatitis B and other immunity-related liver diseases.

Functional Validation of Phytoene Synthase and Lycopene ε-Cyclase Genes for High Lycopene Content in Autumn Olive Fruit (Elaeagnus umbellata).

Lycopene is the most potent antioxidant among all carotenoids and is beneficial to human health. A ripe fruit of autumn olive (Elaeagnus umbellata Thunb.) accumulates a high level of lycopene, which is 5-20 times higher than that in an ordinary tomato fruit. During fruit ripening of autumn olive, only phytoene synthase (EutPSY) expression pattern shows a tight positive correlation with the increased lycopene content observed at four ripening stages, while the lycopene ε-cyclase (EutLCYe) transcript could not be detected throughout fruit ripening. Here, we investigated whether the two genes are important targets for engineering lycopene biosynthesis. The full-length cDNAs of EutPSY and EutLCYe were first isolated. Fruit-specific overexpression of EutPSY in tomato fruits resulted in elevated contents of lycopene and ß-carotene through feedforward regulation of carotenogenic genes, i.e., downregulation of SlLCYe and upregulation of SlLCYb and SlCYCB. These fruits were decreased in ethylene production throughout ripening. Transcript levels of genes for system-2 ethylene synthesis (SlACS2, SlACS4, SlACO1, and SlACO3), perception (SlNR/ETR3 and SlETR4), and response (SlE4 and SlE8) were also inhibited in EutPSY-overexpressing fruits. Repressing ethylene synthesis and signaling transduction delayed fruit climacteric ripening of transgenic tomato plants. Additionally, RNAi suppression of SlLCYe enhanced ß-carotene but not lycopene accumulation through altered expression of carotenogenic genes in transgenic tomato fruits by both feedforward and feedback regulatory mechanisms. Ethylene production in SlLCYe-RNAi fruits decreased, thereby delaying fruit ripening. Collectively, these results confirmed that transcriptional regulation of EutPSY and EutLCYe plays a crucial role and a part in massive lycopene accumulation in autumn olive fruits, respectively. EutPSY overexpression enhanced lycopene accumulation in tomato fruits independently of the ethylene pathway but did not influence the size and weight of tomato fruits. EutPSY can be used as an effective strategy capable of elevating the lycopene content in fruits for improving quality.

Elaeagnus angustifolia Plant Extract Inhibits Epithelial-Mesenchymal Transition and Induces Apoptosis via HER2 Inactivation and JNK Pathway in HER2-Positive Breast Cancer Cells.

Elaeagnus angustifolia (EA) is a medicinal plant used for treating several human diseases in the Middle East. Meanwhile, the outcome of EA extract on HER2-positive breast cancer remains nascent. Thus, we herein investigated the effects of the aqueous EA extract obtained from the flowers of EA on two HER2-positive breast cancer cell lines, SKBR3 and ZR75-1. Our data revealed that EA extract inhibits cell proliferation and deregulates cell-cycle progression of these two cancer cell lines. EA extract also prevents the progression of epithelial-mesenchymal transition (EMT), an important event for cancer invasion and metastasis; this is accompanied by upregulations of E-cadherin and ß-catenin, in addition to downregulations of vimentin and fascin, which are major markers of EMT. Thus, EA extract causes a drastic decrease in cell invasion ability of SKBR3 and ZR75-1 cancer cells. Additionally, we found that EA extract inhibits colony formation of both cell lines in comparison with their matched control. The molecular pathway analysis of HER2 and JNK1/2/3 of EA extract exposed cells revealed that it can block HER2 and JNK1/2/3 activities, which could be the major molecular pathway behind these events. Our findings implicate that EA extract may possess chemo-preventive effects against HER2-positive breast cancer via HER2 inactivation and specifically JNK1/2/3 signaling pathways.

Fermentation with mono- and mixed cultures of Lactobacillus plantarum and L. casei enhances the phytochemical content and biological activities of cherry silverberry (Elaeagnus multiflora Thunb.) fruit.

BACKGROUND: Lactic acid fermentation has been widely used to improve the nutritional and functional properties of food products. Cherry silverberry (Elaeagnus multiflora Thunb.) is considered as an invasive plant species with known medicinal and functional properties. In this study, improvement of the biological activity and health benefits of cherry silverberry fruit through lactic acid fermentation was investigated. RESULTS: Extracts of cherry silverberry fruits fermented by pure cultures of Lactobacillus plantarum KCTC 33131 and L. casei KCTC 13086 exhibited favorable physicochemical properties and enhanced phytochemical content, antioxidant properties (DPPH radical scavenging activity, reducing power, superoxide dismutase-like property and hydrogen peroxide scavenging activity) and α-glucosidase and tyrosinase enzyme inhibitory activity as compared with unfermented fruits. Despite a decrease in the specific phenolic acid contents among the fermented samples, the cherry silverberry fruit fermented by mixed cultures of L. plantarum and L. casei contained superior total polyphenols (3.78 ± 0.22 mg GAE g-1 ) and total (0.66 ± 0.12 mg QE g-1 ) and individual flavonoid contents in comparison with fruits fermented by single cultures and unfermented ones. Multivariate analysis also showed strong association among total phytochemical contents and biological activities. CONCLUSIONS: This work has elucidated the effect of fermentation with L. plantarum KCTC 33131 and L. casei KCTC 13086 on the improvement of the physicochemical properties and biological activity of cherry silverberry fruit. It also revealed the potential application of fermented cherry silverberry in the production of food materials beneficial for health. © 2020 Society of Chemical Industry.

Curative Effect of Catechin Isolated from Elaeagnus Umbellata Thunb. Berries for Diabetes and Related Complications in Streptozotocin-Induced Diabetic Rats Model.

In this study, catechin (CTN) isolated from Elaeagnus umbellata was evaluated for in vitro antioxidant potential and inhibition of carbohydrate digestive enzymes (α-amylase and α-glucosidase). The compound was also tested for its in vivo antidiabetic potential using Sprague-Dawley rats as experimental animals. The effects of various doses of catechin in STZ (Streptozotocin) induced diabetic rats on fasting blood glucose level, body weight, lipid parameters, hepatic enzymes, and renal functions were evaluated using the reported protocols. The CTN exhibited the highest percent antioxidant for free radical scavenging activity against DPPH and ABTS free radicals, and inhibited the activity of carbohydrate digestive enzymes (with percent inhibition values: 79 ± 1.5% α-amylase and 80 ± 1.1% α-glucosidase). Administration CTN and standard glibenclamide significantly decreased the fasting blood glucose level and increased the body weight in STZ-induced diabetic rats. CTN significantly decreased the different lipid parameters, hepatic, and renal function enzyme levels along with Hb1c level in diabetic rats, while significantly increasing the high-density lipoprotein (HDL) level with values comparable to the standard glibenclamide. Further, the altered levels of glutathione and lipid peroxides of liver and kidney tissues were restored (by CTN) to levels similar to the control group. CTN significantly increased the antioxidant enzyme activities, total content of reduced glutathione, and reduced the malondialdehyde (MDA) level in rat liver and kidney tissues homogenates, and also corrected the histopathological abnormalities, suggesting its antioxidant potential.

Proteomics Analysis of E. angustifolia Seedlings Inoculated with Arbuscular Mycorrhizal Fungi under Salt Stress.

To reveal the mechanism of salinity stress alleviation by arbuscular mycorrhizal fungi (AMF), we investigated the growth parameter, soluble sugar, soluble protein, and protein abundance pattern of E. angustifolia seedlings that were cultured under salinity stress (300 mmol/L NaCl) and inoculated by Rhizophagus irregularis (RI). Furthermore, a label-free quantitative proteomics approach was used to reveal the stress-responsive proteins in the leaves of E. angustifolia. The result indicates that the abundance of 75 proteins in the leaves was significantly influenced when E. angustifolia was inoculated with AMF, which were mainly involved in the metabolism, signal transduction, and reactive oxygen species (ROS) scavenging. Furthermore, we identified chorismate mutase, elongation factor mitochondrial, peptidyl-prolyl cis-trans isomerase, calcium-dependent kinase, glutathione S-transferase, glutathione peroxidase, NADH dehydrogenase, alkaline neutral invertase, peroxidase, and other proteins closely related to the salt tolerance process. The proteomic results indicated that E. angustifolia seedlings inoculated with AMF increased the secondary metabolism level of phenylpropane metabolism, enhanced the signal transduction of Ca2+ and ROS scavenging ability, promoted the biosynthesis of protein, accelerated the protein folding, and inhibited the degradation of protein under salt stress. Moreover, AMF enhanced the synthesis of ATP and provided sufficient energy for plant cell activity. This study implied that symbiosis of halophytes and AMF has potential as an application for the improvement of saline-alkali soils.

Examination of Correlation between Histidine and Cadmium Absorption by Eleagnus angustifolia L., Vitis vinifera L. and Nerium oleander L. Using HPLC-MS and ICP-MS.

In this study, HPLC-MS and ICP-MS methods wereused for the determination of histidine and cadmium in Eleagnus angustifolia L., Vitis vinifera L. and Nerium oleander L. leaves taken from industrial area including Gaziantep and Bursa cities. To histidine determination by HPLC-MS, flow rate of mobile phase, fragmentor potential, injection volume and column temperature were optimized as 0.2 mL · min⁻¹, 70 V, 15 µL and 20 °C, respectively. For extraction of histidine from plants, distilled water was used by applying on 90 °C and 30 min. The concentrations (as mg · kg⁻¹) of histidine were found to be in range of 8~22 for Eleagnus angustifolia L., 10~33 for Vitis vinifera L. and 6~11 for Nerium oleander L. The concentrations of cadmium were found to be in ranges of 6~21 µg · kg⁻¹ for Vitis vinifera L. 15~110 µg · kg⁻¹ for Eleagnus angustifolia L. and 63~218 µg · kg⁻¹ for Nerium oleander L.

Proteomic analysis of up-accumulated proteins associated with fruit quality during autumn olive (Elaeagnus umbellata) fruit ripening.

Fruit ripening is a complex phenomenon that makes berries attractive and also determines their nutritional value. Autumn olive ( Elaeagnus umbellata Thunb.) fruit is a rich source of many human health-related nutrients. The changes in pericarp color are initiated at early developmental stages, coinciding with the fast increase in fruit size. Fruit quality traits with special emphasis on soluble sugars, organic acids, lycopene, and total protein contents were assayed during the fruit ripening. In the fully ripe fruit, glucose and fructose were the principal sugars, malic acid was the most abundant organic acid, and lycopene concentration was extremely high. A proteomic analysis was used to identify up-accumulated proteins induced by the ripening. Among 63 up-accumulated protein spots, 43 were successfully identified by MALDI-TOF/TOF-MS. All 43 proteins were novel for autumn olive, and 8 were first reported in the fruit. Twenty-one proteins of known function were involved in sugar metabolism, citric acid cycle, isoprenoid metabolism, fatty acid synthesis, and protein hydrolysis. The possible roles of these 21 accumulated proteins in autumn olive fruit quality are discussed.

Soil and groundwater nitrogen response to invasion by an exotic nitrogen-fixing shrub.

Autumn-olive (Elaeagnus umbellata Thunb.) is an invasive, exotic shrub that has become naturalized in the eastern United States and can fix nitrogen (N) via a symbiotic relationship with the actinomycete Frankia. Fixed N could potentially influence nutrient cycling rates and N leaching into soil water and groundwater. In situ net N mineralization, net nitrification, and net ammonification rates, as well as soil water and groundwater nitrate N (NO(3)-N) and ammonium N (NH(4)-N) concentrations, were measured under autumn-olive-dominated and herbaceous open field areas in southern Illinois. Soil net N mineralization and net nitrification rates were higher under autumn-olive compared with open field (p < 0.05) and could be driven, in part, by the relatively low C/N ratio (11.41 +/- 0.29) of autumn-olive foliage and subsequent litter. Autumn-olive stands also had greater soil water NO(3)-N (p = 0.003), but soil water NH(4)-N concentrations were similar between autumn-olive and open field. Groundwater NO(3)-N and NH(4)-N concentrations were similar beneath both types of vegetation. Groundwater NO(3)-N concentrations did not reflect patterns in soil N mineralization and soil water NO(3)-N most likely due to a weak hydrologic connection between soil water and groundwater. The increased N levels in soil and soil water indicate that abandoned agroecosystems invaded by autumn-olive may be net sources of N to adjacent terrestrial and aquatic systems rather than net sinks.

Nitrogen fixation by Elaeagnus angustifolia in the reclamation of degraded croplands of Central Asia.

Extensive degradation of irrigated croplands, due to increasing soil salinity and depletion of soil nutrient stocks, is a major problem in Central Asia (CA), one of the largest irrigated areas in the world. To assess the potential for improving the productive capacity of degraded lands by afforestation, we examined N(2) fixation of Elaeagnus angustifolia L. in mixed plantations with non-fixing Populus euphratica Oliv. and Ulmus pumila L. Fixation of N(2) was quantified by the (15)N natural abundance technique based on both foliar and whole-plant sampling during five consecutive growing seasons. Despite elevated root-zone soil salinity (6-10 dS m(-1)) and deficiency in plant-available P (4-15 mg kg(-1)), N(2) fixation (%Ndfa) increased from an initial value of 20% to almost 100% over 5 years. Within each growing season, %Ndfa steadily increased and peaked in the fall. Annual N(2) fixation, determined using foliar delta(15)N, initially averaged 0.02 Mg ha(-1), peaked at 0.5 Mg ha(-1) during the next 2 years and thereafter stabilized at 0.3 Mg ha(-1). Estimates based on whole-plant delta(15)N were <10% lower than those based on foliar delta(15)N. The increase in plant-available soil N was significantly higher in E. angustifolia plots than in P. euphratica and U. pumila plots. Increases in the concentrations of organic C (19%), total N (21%) and plant-available P (74%) in the soil were significant irrespective of tree species. This improvement in soil fertility is further evidence that afforestation with mixed-species plantations can be a sustainable land use option for the degraded irrigated croplands in CA.

Natural abundances of 15N and 13C in leaves of some N2-fixing and non-N2-fixing trees and shrubs in Syria.

A survey study was conducted on man-made plantations located at two different areas in the arid region of Syria to determine the variations in natural abundances of the (15)N and (13)C isotopes in leaves of several woody legume and non-legume species, and to better understand the consequence of such variations on nitrogen fixation and carbon assimilation. In the first study area (non-saline soil), the delta(15)N values in four legume species (Acacia cyanophylla,-1.73 per thousand Acacia farnesiana,-0.55 per thousand Prosopis juliflora,-1.64 per thousand; and Medicago arborea,+1.6 \textperthousand) and one actinorhizal plant (Elaeagnus angustifolia,-0.46 to-2.1 per thousand) were found to be close to that of the atmospheric value pointing to a major contribution of N(2) fixing in these species; whereas, delta(15)N values of the non-fixing plant species were highly positive. delta(13)C per thousand; in leaves of the C3 plants were found to be affected by plant species, ranging from a minimum of-28.67 per thousand; to a maximum of-23 per thousand. However, they were relatively similar within each plant species although they were grown at different sites. In the second study area (salt affected soil), a higher carbon discrimination value (Delta(13)C per thousand) was exhibited by P. juliflora, indicating that the latter is a salt tolerant species; however, its delta(15)N was highly positive (+7.03 per thousand) suggesting a negligible contribution of the fixed N(2). Hence, it was concluded that the enhancement of N(2) fixation might be achieved by selection of salt-tolerant Rhizobium strains.

Expression of EuNOD-ARP1 encoding auxin-repressed protein homolog is upregulated by auxin and localized to the fixation zone in root nodules of Elaeagnus umbellata.

Root nodule formation is controlled by plant hormones such as auxin. Auxin-repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD-ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD-ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules.

Intra-specific genetic relationship analyses of Elaeagnus angustifolia based on RP-HPLC biochemical markers.

Elaeagnus angustifolia Linn. has various ecological, medicinal and economical uses. An approach was established using RP-HPLC (reversed-phase high-performance liquid chromatography) to classify and analyse the intra-specific genetic relationships of seventeen populations of E. angustifolia, collected from the Xinjiang areas of China. Chromatograms of alcohol-soluble proteins produced by seventeen populations of E. angustifolia, were compared. Each chromatogram of alcohol-soluble proteins came from a single seed of one wild plant only. The results showed that when using a Waters Delta Pak. C18, 5 microm particle size reversed phase column (150 mm x 3.9 mm), a linear gradient of 25%-60% solvent B with flow rate of 1 ml/min and run time of 67 min, the chromatography yielded optimum separation of E. angustifolia alcohol-soluble proteins. Representative peaks in each population were chosen according to peak area and occurrence in every seed. The converted data on the elution peaks of each population were different and could be used to represent those populations. GSC (genetic similarity coefficients) of 41% to 62% showed a medium degree of genetic diversity among the populations in these eco-areas. Cluster analysis showed that the seventeen populations of E. angustifolia could be divided into six clusters at the GSC=0.535 level and indicated the general and unique biochemical markers of these clusters. We suggest that E. angustifolia distribution in these eco-areas could be classified into six variable species. RP-HPLC was shown to be a rapid, repeatable and reliable method for E. angustifolia classification and identification and for analysis of genetic diversity.