An overview of Parafrankia (Nod+/Fix+) and Pseudofrankia (Nod+/Fix-) interactions through genome mining and experimental modeling in co-culture and co-inoculation of Elaeagnus angustifolia.

In many frankia, the ability to nodulate host plants (Nod+) and fix nitrogen (Fix+) is a common strategy. However, some frankia within the Pseudofrankia genus lack one or two of these traits. This phenomenon has been consistently observed across various actinorhizal nodule isolates, displaying Nod- and/or Fix- phenotypes. Yet, the mechanisms supporting the colonization and persistence of these inefficient frankia within nodules, both with and without symbiotic strains (Nod+/Fix+), remain unclear. It is also uncertain whether these associations burden or benefit host plants. This study delves into the ecological interactions between Parafrankia EUN1f and Pseudofrankia inefficax EuI1c, isolated from Elaeagnus umbellata nodules. EUN1f (Nod+/Fix+) and EuI1c (Nod+/Fix-) display contrasting symbiotic traits. While the prediction suggests a competitive scenario, the absence of direct interaction evidence implies that the competitive advantage of EUN1f and EuI1c is likely contingent on contextual factors such as substrate availability and the specific nature of stressors in their respective habitats. In co-culture, EUN1f outperforms EuI1c, especially under specific conditions, driven by its nitrogenase activity. Iron-depleted conditions favor EUN1f, emphasizing iron's role in microbial competition. Both strains benefit from host root exudates in pure culture, but EUN1f dominates in co-culture, enhancing its competitive traits. Nodulation experiments show that host plant preferences align with inoculum strain abundance under nitrogen-depleted conditions, while consistently favoring EUN1f in nitrogen-supplied media. This study unveils competitive dynamics and niche exclusion between EUN1f and EuI1c, suggesting that host plant may penalize less effective strains and even all strains. These findings highlight the complex interplay between strain competition and host selective pressure, warranting further research into the underlying mechanisms shaping plant-microbe-microbe interactions in diverse ecosystems. IMPORTANCE: While Pseudofrankia strains typically lack the common traits of ability to nodulate the host plant (Nod-) and/or fix nitrogen (Fix-), they are still recovered from actinorhizal nodules. The enigmatic question of how and why these unconventional strains establish themselves within nodule tissue, thriving either alongside symbiotic strains (Nod+/Fix+) or independently, while considering potential metabolic costs to the host plant, remains a perplexing puzzle. This study endeavors to unravel the competitive dynamics between Pseudofrankia inefficax strain EuI1c (Nod+/Fix-) and Parafrankia strain EU1Nf (Nod+/Fix+) through a comprehensive exploration of genomic data and empirical modeling, conducted both in controlled laboratory settings and within the host plant environment.

Green synthesis of nanohydroxyapatite trough Elaeagnus angustifolia L. extract and evaluating its anti-tumor properties in MCF7 breast cancer cell line.

BACKGROUND: One of the most common types of cancer in women is breast cancer. There are numerous natural plant-based products, which exert anti-tumoral effects including Elaeagnus Angustifolia (EA). It modulates cell-cycle process, heat-shock proteins expression, anti-proliferative properties, apoptosis induction, blocking of angiogenesis, and cell invasion inhibition. The current study aimed to synthesize and evaluate the anticancer effects of hydroalcoholic EA extract (HEAE), Nanohydroxyapatite (nHAp) and nHAp synthesized trough EA (nHA-EA) in MCF-7 breast cancer cell line. METHODS: In the present study, HEAE preparation and green synthesis of nHA-EA was done and phase composition, functional groups, and crystallin phase of nHA-EA and nHAp were determined using Fourier-transform infrared (FTIR) and X-ray diffraction (XRD). The characteristics of synthesized nanoparticles including structural and morphological parameters were investigated using scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) techniques. Then, by using MTT-assay (Dimethylthiazoldiphenyltetrazolium), the in vitro cytotoxic and half maximal inhibitory concentration (IC50) of EA extract, nHAp, and nHA-EA in the MCF-7 breast cancer cell line was evaluated. Next, we assessed the expression of apoptosis-related genes Bax, Bcl2 and p53 using quantitative reverse-transcriptase polymerase-chain-reaction (qRT-PCR) and migration of MCF-7 cells by scratch assay. RESULTS: The FTIR results demonstrated formation of nHAp and its interaction with HEAE during synthesis process. The XRD results of the synthesized nanoparticles showed similar XRD pattern of nHA-EA and nHAp and purity of synthesized nanomaterials. The average IC50 of HEAE, nHAp, and nHA-EA extract after treatment of cancer cells for 24 h was 400 µg/mL, 200 µg/mL, and 100 µg/mL, respectively. Our results revealed that nHA-EA significantly reduced the migration and invasion of the MCF-7 cells, in comparison to the nHAp and EA extract. Moreover, level of Bax/Bcl2 and p53 was significantly higher in the nHA-EA extract group in comparison to the EA extract and nHAp group. CONCLUSION: Taken together, our results demonstrated that bioactive constituents of EA medicinal plant in form of nHA-EA particles, can effectively exerts potential anticancer and chemo preventive effect against breast cancer growth and can be proposed as a promising beneficial candidate for BC therapy. However, further investigations are required to discover what bioactive compounds are responsible for the chemo preventive effect of this extract.

Isolation, Structural Characterization, and Biological Activity of the Two Acidic Polysaccharides from the Fruits of the Elaeagnus angustifolia Linnaeus.

Elaeagnus angustifolia Linnaeus is a medicinal plant and its fruit has pharmacological activity such as antiinflammatory, antiedema, antinociceptive, and muscle relaxant functions, etc. Two acidic homogeneous polysaccharides (EAP-H-a1 and EAP-H-a2) were isolated from the fruits of Elaeagnus angustifolia L. through DEAE-52 and Sephadex G-75 column chromatography, and the physicochemical, structural properties, and biological activities of the polysaccharides were investigated. Both EAP-H-a1 and EAP-H-a2 were composed of Rha, Ara, Xyl, Glc, and Gal with the molar ratios of 13.7:20.5:23.3:8.8:33.4 and 24.8:19.7:8.2:8.4:38.6, respectively, and with the molecular weights of 705.796 kDa and 439.852 kDa, respectively. The results obtained from Fourier transform infrared spectroscopy (FTIR) confirmed the polysaccharide nature of the isolated substances. Congo red assay confirmed the existence of a triple-helix structure. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis revealed that EAP-H-a1 and EAP-H-a2 had irregular fibrous, filament-like surfaces; and both had crystalline and amorphous structures. Bioactivity analysis showed that the crude polysaccharide, EAP-H-a1, and EAP-H-a2 had clear DPPH and ABTS free radical scavenging activity, and could promote the secretion of NO and the phagocytic activities of RAW 264.7 and THP cells, which showed clear antioxidant and immuno-regulatory activity. These results indicated that Elaeagnus angustifolia L fruit acidic polysaccharides may have potential value in the pharmaceutical and functional food industries.

Antibacterial, Antioxidant, and Phytotoxic Potential of Phytosynthesized Silver Nanoparticles Using Elaeagnus umbellata Fruit Extract.

Due to its eco-friendliness, cost-effectiveness, ability to be handled safely, and a wide variety of biological activities, the green plant-mediated synthesis of nanoparticles has become increasingly popular. The present work deals with the green synthesis and characterization of silver nanoparticles (AgNPs) using Elaeagnus umbellata (fruit) and the evaluation of its antibacterial, antioxidant, and phytotoxic activities. For the synthesis of AgNPs, fruit extract was treated with a 4 mM AgNO3 solution at room temperature, and a color change was observed. In UV-Visible spectroscopy, an absorption peak formation at 456 nm was the sign that AgNPs were present in the reaction solution. Scanning electron microscopy and physicochemical X-ray diffraction were used to characterize AgNPs, which revealed that they were crystalline, spherical, and had an average size of 11.94 ± 7.325 nm. The synthesized AgNPs showed excellent antibacterial activity against Klebsiella pneumoniae (14 mm), Staphylococcus aureus (13.5 mm), Proteus mirabilis (13 mm), and Pseudomonas aeruginosa (12.5 mm), as well as considerable antioxidant activity against DPPH with 69% inhibition at an IC50 value of 43.38 µg/mL. AgNPs also exhibited a concentration-dependent effect on rice plants. Root and shoot length were found to be positively impacted at all concentrations, i.e., 12.5 µg/mL, 25 µg/mL, 50 µg/mL, and 100 µg/mL. Among these concentrations, the 50 µg/mL concentration of AgNPs was found to be most effective. The plant biomass decreased at higher AgNP exposure levels (i.e., 100 µg/mL), whereas 50 µg/mL caused a significant increase in plant biomass as compared to the control. This study provides an eco-friendly method for the synthesis of AgNPs which can be used for their antibacterial and antioxidant activities and also as growth promoters of crop plants.

Anti-inflammatory effects of N-Acetylcysteine and Elaeagnus angustifolia extract on acute lung injury induced by λ-carrageenan in rat.

N-Acetylcysteine (NAC) is a chemical compound with anti-inflammatory and antioxidant activity and acts as a free radical scavenger. Elaeagnus angustifolia (EA) is a plant native to the western part of Iran, with antioxidant and anti-inflammatory properties. The present study been taken evaluated the protective effect afforded by EA and NAC extracts on carrageenan-induced acute lung injury in Wistar rats. In this study, 42 rats were randomly assigned into seven groups. NAC and EA extracts were orally administered once/day for 21 continuous days. Pulmonary damage was induced by intratracheal injection of 100 µl of 2% λ-Carrageenan on day 21. Twenty-four hours post-surgery, the rats were euthanized and the samples were collected. Pretreatment with NAC and EA extracts reduced the total and differential cell accumulation as well as IL-6, and TNF-α cytokines. Antioxidant indicators demonstrate that in the groups receiving NAC and EA extract, MDA decreased while thiol and antioxidant capacity elevated. Treatment with NAC and EA significantly reduced Carrageenan-induced pathological pulmonary tissue injury. NAC and EA extract has protective effects on acute carrageenan-induced lung injury.

The Bioactive Profile, Nutritional Value, Health Benefits and Agronomic Requirements of Cherry Silverberry (Elaeagnus multiflora Thunb.): A Review.

The cherry silverberry (Elaeagnus multiflora Thunb.) is a lesser-known plant species with high nutritional and therapeutic potential. Cherry silverberry contains numerous biologically active compounds. The cherry silverberry is a shrub growing up to 3 m. Its drupe-like fruit is ellipsoidal, up to 1 cm long, and set on stems. It is red in color, juicy, and sour, and its taste resembles that of red currants. According to the literature, cherry silverberry fruit contains carbohydrates, organic acids, and amino acids, as well as vitamin C, in addition to biominerals, polyphenols, flavonoids, carotenoids, chlorophylls, and tocopherols, which contribute to its high nutritional value. New biotypes of cherry silverberry cultivated in Poland can be used for the production of functional foods and direct consumption. In China, the cherry silverberry, known as goumi, has been used as a medicinal plant and a natural remedy for cough, diarrhea, itch, foul sores, and, even, cancer. This review article summarizes the scant research findings on the nutritional and therapeutic benefits of cherry silverberry.

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.

Changes in Physicochemical, Free Radical Activity, Total Phenolic and Sensory Properties of Orange (Citrus sinensis L.) Juice Fortified with Different Oleaster (Elaeagnus angustifolia L.) Extracts.

In Iran and other parts of Western Asia, the oleaster (Elaeagnus angustifolia L.) fruit is processed in the dried powdery form, and in recent times, increasingly applied/sprinkled in fruit juices such as those made from oranges (Citrus sinensis L.). To our best knowledge, the effectiveness of oleaster fruit extract in fortifying the orange juice has not yet been reported and the knowledge of this will greatly benefit the consumers, particularly those around the Western Asia region. This current work, therefore, investigated the changes in physicochemical, free radical activity, total phenolic compounds, and sensory properties of orange juice fortified with different oleaster fruit extracts. The orange juice mix formulation comprised different concentrations (5, 10, 15, 20, and 25%) of oleaster (alcoholic, aqueous, and hydro-alcoholic) extracts. The control comprised orange concentrate (4% w/v), sugar (8.5% w/v), and citric acid (0.1% w/v) brought to the desirable volume with water. As the free radical activity depicted the antioxidant properties, the physicochemical aspects of this work involved the determinations of Brix, density, ash, pH, total acidity, sucrose, and total sugar, whereas the sensory aspects involved the determinations of color and taste. Whilst the aqueous oleaster 20 and 25% extracts produced notable physicochemical differences in the orange juice mix, both free radical activity, and phenolic compounds significantly increased (p < 0.05) after 30 days despite resembling (p > 0.05) those of control at day 1. More so, the increases in aqueous, alcoholic, and hydro-alcoholic oleaster extracts would decrease (p < 0.05) the sensory color and taste of the orange juice mix in this study.

Green synthesis of zinc oxide nanoparticles using Elaeagnus angustifolia L. leaf extracts and their multiple in vitro biological applications.

Due to their versatile applications, ZnONPs have been formulated by several approaches, including green chemistry methods. In the current study, convenient and economically viable ZnONPs were produced using Elaeagnus angustifolia (EA) leaf extracts. The phytochemicals from E. angustifolia L. are believed to serve as a non-toxic source of reducing and stabilizing agents. The physical and chemical properties of ZnONPs were investigated employing varying analytical techniques (UV, XRD, FT-IR, EDX, SEM, TEM, DLS and Raman). Strong UV-Vis absorption at 399 nm was observed for green ZnONPs. TEM, SEM and XRD analyses determined the nanoscale size, morphology and crystalline structure of ZnONPs, respectively. The ZnONPs were substantiated by evaluation using HepG2 (IC50: 21.7 µg mL-1) and HUH7 (IC50: 29.8 µg mL-1) cancer cell lines and displayed potential anticancer activities. The MTT cytotoxicity assay was conducted using Leishmania tropica "KWH23" (promastigotes: IC50, 24.9 µg mL-1; and amastigotes: IC50, 32.83 µg mL-1). ZnONPs exhibited excellent antimicrobial potencies against five different bacterial and fungal species via the disc-diffusion method, and their MIC values were calculated. ZnONPs were found to be biocompatible using human erythrocytes and macrophages. Free radical scavenging tests revealed excellent antioxidant activities. Enzyme inhibition assays were performed and revealed excellent potential. These findings suggested that EA@ZnONPs have potential applications and could be used as a promising candidate for clinical development.

Antioxidants Isolated from Elaeagnus umbellata (Thunb.) Protect against Bacterial Infections and Diabetes in Streptozotocin-Induced Diabetic Rat Model.

The increase in resistance of microbes against conventional drugs is currently a hot issue, whereas diabetes is another main cause of mortalities encountered throughout the world after cancer and heart attacks. New drug sources in the form of plants are investigated to get effective drugs for the mentioned diseases with minimum side effects. Elaeagnus umbellata Thunb. is a medicinal plant used for the management of stress related disorders like diabetes and other health complications. The active constituents of the chloroform extract derived from E. umbellata berries was isolated by silica gel column chromatography which were identified as morin, phloroglucinol, and 1-hexyl benzene through various spectroscopic techniques (electron ionization mass spectrometry, 1H-NMR, and 13C-NMR spectroscopy). The possible protective effects (antioxidant, antibacterial, and antidiabetic activity) of isolated compounds were evaluated using reported methods. Morin exhibited strong in vitro antiradical potential against DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) radicals along with prominent antibacterial activities against selected bacterial strains (Escherichia coli, Bacillus cereus, Salmonella typhi, Klebsiella pneumonia, Pseudomonas aeruginosa and Proteus mirabilis). Among the isolated compounds the more potent one (morin) was assessed for its in vivo antidiabetic potential in streptozotocin-induced diabetic rat model. The in vivo effects observed were further confirmed in ex vivo experiments where the effect of isolated compound on antioxidant enzyme like glutathione peroxidase (GPx), total content of reduced glutathione (GSH), % DPPH inhibition, and the lipid peroxidation MDA (Malondialdehyde) level in pancreatic tissues homogenates were evaluated. In vivo morin at tested doses (2, 10, 15, 30 and 50 mg/kg body weight) significantly restored the alterations in the levels of fasting blood glucose level and body weight loss along with significant decrease in levels of cholesterol, triglycerides, low density lipoprotein, HbA1c level, and significantly increased the high-density lipoprotein in diabetic rats. Morin also effectively ameliorated the hepatic enzymes, and renal functions like serum creatinine. Morin significantly increased the antioxidant enzyme like GPx activity, GSH content, and % DPPH inhibition activity, while reduced the lipid peroxidation MDA (malondialdehyde) level in pancreatic tissues homogenates, and modification of histopathological changes in diabetic rats. Morin exhibited high antioxidant, antibacterial, and antidiabetic potentials as compared to phloroglucinol and 1-hexyl benzene, that could, therefore, be considered as a promising therapeutic agent to treat diabetes mellitus and bacterial infections.

trans-Tiliroside: A potent α-glucosidase inhibitor from the leaves of Elaeagnus angustifolia L.

Elaeagnus angustifolia L. (Elaeagnaceae) is an important medicinal plant associated with numerous pharmacological activities. Its leaves are used as a therapeutic agent in traditional medicinal systems to treat diabetes. However, the active compounds responsible for the beneficial effects of E. angustifolia remain unclear. In this study, we determined the bioactive profile of E. angustifolia leaves using open column chromatography and semi-preparative HPLC. Further, we sought to determine its α-glucosidase and α-amylase inhibitory activities, and its DPPH and ABTS radical-scavenging activities. Four undescribed flavonol glycosides, igdoside A-D, and four known glucosides were isolated from the ethyl acetate and n-butanol extracts of E. angustifolia leaves. Thereafter, the compound structures were identified using spectroscopic methods, including NMR and mass spectrometry. Of the compounds extracted, kaempferol-3-O-(6″-trans-p-coumaroyl)-ß-D-glucopyranoside (trans-tiliroside), exhibited the highest α-glucosidase inhibitory activity with an IC50 value of 2128 ±â€¯63 µM compared to the positive control, acarbose (IC50 = 6561 ±â€¯207 µM). trans-Tiliroside was also found to exhibit potent scavenging activity against the ABTS radical, with an IC50 value of 5 ±â€¯0 µM, compared to the positive controls, trolox (31 ±â€¯1 µM) and α-tocopherol (50 ±â€¯1 µM). In addition, isorhamnetin-3-O-ß-D-galactopyranoside (IC50 = 6 ±â€¯0 µM) and astragalin (IC50 = 6 ±â€¯0 µM) showed similar ABTS radical-scavenging activity as trans-tiliroside. Based on HPLC, the content of trans-tiliroside was 9.69% in the ethyl acetate extract, 1.04% in decoction, 0.34% in 70% methanol extract, and 0.23% in infusion. None of the extracts and compounds showed α-amylase inhibition or DPPH-scavenging activity.

Anti-ulcerogenic effect of methanolic extract of Elaeagnus conferta Roxb. seeds in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Elaeagnus conferta Roxb. (Elaeagnaceae) is a subtropical shrub mainly native to India, Vietnam, Malaysia and South China, whose various parts are used for treatment of diabetes, gastric ulcers, pain, oxidative stress and pulmonary disorders. Though the other parts of the plant have been reported for their ethnic use i.e. fruits as astringent locally and for cancer systemically, leaves for body pain and flowers for pain in chest and the seeds are mentioned as edible, there is no report per se on the medicinal use of seeds. Based on the fact that seeds of closely resembling species i.e. Elaeagnus rhamnoides has demonstrated significant anti-gastroulcerative property, the probability of the seeds of E. conferta possessing similar activity seemed quite significant. AIM OF THE STUDY: Phytochemical investigation and assessment of pharmacological mechanism(s) involved in anti-ulcer effect of methanolic extract of the seeds of E. conferta. MATERIALS AND METHODS: Bioactive phytoconstituents were isolated by column chromatography. These were identified by spectroscopic techniques including infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) and mass spectrometry. Methanolic extract (MEC) of the seeds was prepared by cold maceration and its anti-ulcerogenic potential was evaluated using indomethacin (50 mg/kg) and water immersion stress models in male rats. The animals were pre-treated with different doses of MEC (400 and 800 mg/kg) and the therapeutic effect was compared with standard drug i.e. ranitidine (RANT; 50 mg/kg). The ameliorative effects of MEC were investigated on gastric juice pH, total acidity, free acidity and ulcer index. The assays of malionaldehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and pro-inflammatory cytokines i.e. interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were carried out to find out the possible mechanism(s) of protection. Further, histopathological changes were also studied. RESULTS: Chromatography studies and further confirmation by spectroscopic techniques revealed the presence of four different compounds in MEC i.e oleic acid (1), stearic acid (2), ascorbic acid (3) and quercetin (4). MEC exhibited anti-ulcerogenic effect in dose dependent manner which may be attributed to suppression of pro-inflammatory cytokines (IL-6, TNF-α) and MDA (112.7%), and up-regulation of protective factors such as CAT (90.48%), SOD (92.77%) and GSH (90.01%). Ulcer inhibition, reduction in total and free acidity and increase in gastric juice pH were observed in MEC treated rats as compared to disease control animals. Histopathological findings confirmed decreased cell infiltration, less epithelial cell damage and regeneration of gastric mucosa in dose dependent manner. CONCLUSIONS: The anti-ulcer effect of MEC may be attributed to its ability to scavenge free radicals and anti-inflammatory property via suppression of TNF-α and IL-6, thus offers a complete and holistic approach for management of peptic ulcer.

In vitro and in vivo relaxation and anti-inflammation of natural flavonoids from Elaeagnus pungens leaf via L-type calcium channel and targeting MAPK signal pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Chinese medicine (TCM), the leaf of Elaeagnus pungens Thunb. (Family Elaeagnaceae) is a herb documented as an antiasthmatic remedy to treat the severe asthma, bronchitis and other respiratory diseases in the early material medica "Bencao Gangmu" (Ming dynasty, about 442 years ago). AIM OF THE STUDY: This work is purposed to investigate the pharmacological effects and mechanism of total flavonoids from Elaeagnus pungens leaves (FLA) on asthma in vivo and vitro. MATERIALS AND METHODS: Female BALB/c mice were sensitized by intraperitoneal injection of OVA with aluminum hydroxide and intranasal challenged with OVA. After treatment with FLA (10, 20 mg/kg p.o.), the behaviors of mice were observed by score evaluation. Enumeration of total cells and OVA-specific IgE assay in the blood were measured as well as enumeration of total cells and cytokines assay in the BALF. Furthermore, histopathological analysis was performed by HE staining. The in vitro relaxing action on muscle force of FLA (0.0316-10.0 mg/ml) was evaluated using isometric tension in tracheal rings, and VDLCC currents were recorded to explore the relaxation mechanism in the isolated tracheal rings and mouse ASM cells, respectively. In vitro anti-inflammatory actions were assessed with LPS-stimulated RAW 264.7 macrophages. The production of inflammatory mediators and MAPK signaling pathway was estimated using ELISA and Western blotting analysis, respectively. RESULTS: The high dose of flavones from E. pungens leaf (20 mg/kg) can significantly improve the symptom of asthma breakout and relieve the lung swelling. FLA treatment decreased eosinophils and leukocytes numbers in blood and BLAF with a dosedependent manner. Furthermore, the inhibiting effect of FLA on the level of Ig E and inflammatory-related cytokines including TNF-α, IL-5 showed dose-independent. FLA relaxed high K + -induced contraction in a dose-dependent manner. The maximal relaxation produced by FLA was 99.7% (IC 50 = 0.46 mg/ml). The whole-cell VDLCC currents were abolished by FLA (3.16 mg/ml) and FLA significantly decreased the maximal amplitude of VDLCCs. No cytotoxic effect of FLA was observed in RAW264.7 cells under the tested concentrations (1-300 µg/mL). The increased IL-6 and NO by the stimulation of LPS in RAW264.7 cells were significantly inhibited by FLA in the dosedependent manner. Treatment with LPS in the presence of FLA, LPS-induced phosphorylation of ERK1/2 and JNK was inhibited in the macrophages. CONCLUSION: FLA from Elaeagnus pungens leaf can alleviate the inflammation symptom via reducing the eosinophils and leukocytes numbers as well as the production of pro-inflammatory cytokines. This anti-inflammatory effect is related to the modulation of the MAPK signaling pathway. FLA can relax the precontracted TRs by blocking the VDLCCs, which interrupts extracellular Ca 2+ influx and inhibit the rise of [Ca 2+ ]i. It strongly suggests that these flavonoids components are the substances basis of Elaeagnus pungens leaves for allergic action, bronchospasm and inflammation in asthma.

Anti-Platelet Properties of Phenolic and Nonpolar Fractions Isolated from Various Organs of Elaeagnus rhamnoides (L.) A. Nelson in Whole Blood.

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a shrub growing in coastal areas. Its organs contain a range of bioactive substances including vitamins, fatty acids, various micro and macro elements, as well as phenolic compounds. Numerous studies of sea buckthorn have found it to have anticancer, anti-ulcer, hepatoprotective, antibacterial, and antiviral properties. Some studies suggest that it also affects the hemostasis system. The aim of the study was to determine the effect of six polyphenols rich and triterpenic acids rich fractions (A-F), taken from various organs of sea buckthorn, on the activation of blood platelets using whole blood, and to assess the effect of the tested fractions on platelet proteins: fraction A (polyphenols rich fraction from fruits), fraction B (triterpenic acids rich fraction from fruits), fraction C (polyphenols rich fraction from leaves), fraction D (triterpenic acids rich fraction from leaves), fraction E (polyphenols rich fraction from twigs), and fraction F (triterpenic acids rich fraction from twigs). Hemostasis parameters were determined using flow cytometry and T-TAS (Total Thrombus-formation Analysis System). Additionally, electrophoresis was performed under reducing and non-reducing conditions. Although all tested fractions inhibit platelet activation, the greatest anti-platelet activity was demonstrated by fraction A, which was rich in flavonol glycosides. In addition, none of the tested fractions (A-F) caused any changes in the platelet proteome, and their anti-platelet potential is not dependent on the P2Y12 receptor.

Inhibitory Effect of Elaeagnus umbellata Fractions on Melanogenesis in α-MSH-Stimulated B16-F10 Melanoma Cells.

When skin is exposed to UV radiation, melanocytes produce melanin. Excessive melanin production leads to skin pigmentation, which causes various cosmetic and health problems. Therefore, the development of safe, natural therapeutics that inhibit the production of melanin is necessary. Elaeagnus umbellata (EU) has long been widely used as a folk medicinal plant because of pharmacological properties that include anti-ulcer, antioxidant, and anti-inflammatory properties. In this study, we investigated the antioxidant activity and melanogenesis inhibitory effects of EU fractions in B16-F10 melanoma cells. EU fractions showed a dose-dependent increase in antioxidant activity in radical scavenging activity. In addition, we evaluated the effect of EU fractions on tyrosinase activity and melanogenesis in α-melanocyte-stimulating hormone-induced B16-F10 melanoma cells. EU was noncytotoxic at 12.5-50 µg/mL. EU fractions effectively inhibited tyrosinase activity and melanogenesis, suppressed the phosphorylation of CREB and ERK involved in the melanogenesis pathway, and down-regulated expression of melanogenesis-related proteins. Interestingly, the anti-melanogenesis effect was most effective at a concentration of 50 µg/mL EU, and the effects of the fractions were superior to those of the extract. Therefore, our study suggests that EU has potential as a safe treatment for excessive pigmentation or as a natural ingredient in cosmetics.

Multidirectional effects of saponin fraction isolated from the leaves of sea buckthorn Elaeagnus rhamnoides (L.) A. Nelson.

Many studies show that saponins isolated from various plants have a cytotoxic effect on cancer cells inducing apoptosis and autophagy. On the other hand, saponins also exhibit a number of beneficial properties, such as antioxidant properties. Thus, saponins can be considered both in terms of their therapeutic and protective effects during anticancer treatment. In this study, we investigated the effect of the saponin fraction isolated from sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) leaves on the viability of HL-60 cancer cells using resazurin assay and its ability to induction of apoptosis with Annexin V-FITC and propidium iodide (PI) double staining. Moreover, we studied its effect on the oxidative stress induced by H2O2, and anti-platelet and anticoagulant potential in whole blood using T-TAS, a microchip-based flow chamber system. We observed that the saponin fraction significantly decreased the viability of HL-60 cells at the concentration above 50 µg/mL and induced apoptosis at the concentration of 100 µg/mL. Moreover, we observed that saponin fraction used at lower concentrations, such as 0.5 and 1 µg/mL, stimulated HL-60 cells and increased their viability. The saponin fraction also decreased the level of free radicals and reduced oxidative DNA damage measured by the comet assay. However, at high concentration of oxidant H2O2 equal 5 mM, we noticed that the saponin fraction at 50 µg/mL increased the level of free radicals in HL-60 cells. We also demonstrated anticoagulant potential of the saponin fraction at the concentration of 50 µg/mL. Our results indicate that the saponin fraction obtained from sea buckthorn leaves can show both chemotherapeutic and chemoprotective potential.

Chemical composition, in vitro antioxidant, anticholinesterase, and antidiabetic potential of essential oil of Elaeagnus umbellata Thunb.

BACKGROUND: Elaeagnus umbellata Thunb. (autumn olive) is a high valued medicinal plant. It belongs to Elaeagnaceae family and is widely distributed in Himalayan regions of Pakistan. In the present study essential oil were extracted from the fruit of this plant and their antioxidant, anticholinesterase and antidiabetic potentials were also evaluated. METHODS: Essential oils were extracted from the fruit of E. umbellata using hydro-distillation method and were characterized by GC-MS. The extracted oil were tested for its antioxidant, anticholinesterase, and antidiabetic potentials using standard protocols. RESULTS: About 68 compounds were identified by GC-MS. The extracted oil exhibited a fairly high free radical scavenging activities against DPPH and ABTS radicals with IC50 values of 70 and 105 µg/mL respectively (for ascorbic acid, used as standard, the IC50 values were 32 and 29 µg/mL, respectively against the mentioned radicals). The essential oil also exhibited anticholinesterase activities with IC50 values of 48 and 90 µg/mL respectively against AChE and BChE (for galantamine used as standard, the IC50 values were 25 and 30 µg/mL respectively). The essential oil also exhibited antidiabetic potential with IC50 values of 120 and 110 µg/mL respectively against α-glucosidase and α-amylase (IC50 values for standard acarbose = 28 and 30 µg/mL respectively). CONCLUSION: Essential oil extracted from the fruits of E. umbellata exhibited reasonable antioxidant, anticholinesterase, and antidiabetic potentials that could be used as alternative medicine in treating diabetes and neurodegenerative disorders. However, further studies are needed to isolate responsible compounds and evaluate the observed potential in animal models.

Novel bioactive properties of low-polarity fractions from sea-buckthorn extracts (Elaeagnus rhamnoides (L.) A. Nelson) - (in vitro).

Due to the richness of bioactive substances and easy accessibility, sea-buckthorn can be an ingredient of currently popular functional food supporting anti-cancer therapy. Low-polarity fractions from fruit (OL), twigs (GL) and leaves (LL) were investigated. Compared to the previous scientific reports a more detailed analysis of the chemical composition of individual fractions was performed. Cytotoxicity of low-polarity fractions has been investigated and activity compared in human tumor and normal cells cultured in vitro. The genotoxicity and pro-apoptotic properties of low-polarity fractions were also followed on selected cell lines that had proved to be the most sensitive. In the proposed research model being tested, low-polarity fractions act cytotoxically, even 3 times more strongly in cancer cells than normal ones. Measurement of caspase 3/7 activity indicated that cell death occurs through apoptosis. Furthermore, high concentrations of low-polarity fractions have moderate genotoxic properties. Data obtained on the biological properties of low-polarity fractions from sea-buckthorn show that these fractions can potentially support cancer cells elimination. Phytotochemical analysis indicates the key role of the triterpenoids in this process.

The Oleaster (Elaeagnus angustifolia): A Comprehensive Review on Its Composition, Ethnobotanical and Prebiotic Values>.

BACKGROUND: Oleaster or Elaeagnus angustifolia is a deciduous plant from Elaegnacea family and is well-known for its remedial applications. OBJECTIVE: This paper presents a comprehensive review of the potential application of Oleaster's flour incorporated in some food products. Emphasis is given to the physicochemical, biochemical, and functional properties of Oleaster's flour. METHODS: A comprehensive search was carried out to find publications on Oleaster's flour and its application as a prebiotic. The results of the related studies were extracted and summarized in this paper. RESULTS: Oleaster's flour as a prebiotic ingredient enhances antioxidants, polyphenols, fiber, flavonoids, Sterols, carbohydrates, and protein content of food products. CONCLUSION: Further advanced investigations on Oleaster and its functional ingredients revealed that these are efficacious and can be applied as a substitute source in pharmacological industries for medical applications.

The effects of Elaeagnus angustifolia L. on lipid and glycaemic profiles and cardiovascular function in menopausal women: A double-blind, randomized, placebo-controlled study.

AIMS: The reduced production of ovarian hormones is considered to be the cause of an increase in the incidence of heart disease in women after menopause. Phytoestrogens are found in various herbal sources and are considered as an alternative to hormone therapy because of structural similarity with oestrogen. Elaeagnus angustifolia L., known as Senjed in Persian, is used in Iranian traditional medicine with various medicinal properties, contains valuable compounds, including two types of phytoestrogens. The aim of the present study was to investigate the efficacy of E. angustifolia fruit on the cardiovascular function, lipid and glycaemic profiles in postmenopausal women. METHODS: In this double-blind placebo-controlled clinical trial, 58 postmenopausal women were randomly assigned into two experimental groups of medicinal herb (15 g/day of the whole E. angustifolia fruit powder) and placebo (15 g/day of isomalt + corn starch). Before the trial and after 10 weeks of the treatment, cardiovascular function (heart rate, blood pressure), serum glycaemic profile (fasting blood glucose; glycated haemoglobin, HbA1C), insulin and lipid profile (total cholesterol, TC; triglyceride, TG; LDL-C and HDL-C) were measured. RESULTS: Heart rate, and serum level of LDL-C and HDL-C significantly decreased after treatment with E. angustifolia. Changes in glycaemic profile were not clinically significant. In addition, some studied biochemical parameters significantly changed in the placebo group. CONCLUSIONS: E. angustifolia was somewhat effective in improving cardiovascular function and lipid profile, as well as the overall health of postmenopausal women. However, the use of isomalt and corn starch in menopausal women needs further investigation.