Natural Antioxidant Flavonoid Research Articles

Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function

This pioneering research investigated apigenin potential to augment rooster sperm cryosurvival in an extender model. Apigenin is a natural antioxidant flavonoid showing promise for improved post-thaw sperm function. However, its effects on avian semen cryopreservation remain unexplored. This first study supplemented rooster sperm Lake extender with 0, 50, 100, 200, 400 μmol/L apigenin to determine the optimal concentrations for post-thaw quality. Supplementation with 100 μmol/L apigenin resulted in significant enhancements in total motility (from 41.5% up to 71.5%), progressive motility (18.1% to 29.1%) (p < 0.05), membrane integrity (40% to 68%), mitochondrial function (p < 0.001), viability (37% to 62%) and total antioxidant capacity (p < 0.001) compared to the control. It also substantially reduced percentages of abnormal morphology, reactive oxygen species and apoptosis (p < 0.001). Although 200 μmol/L apigenin significantly enhanced some attributes, effects were markedly lower than 100 μmol/L. Higher doses did not improve cryoprotective parameters. This indicates 100 μmol/L as the optimal apigenin concentration. This represents the first report of apigenin protecting rooster sperm from cryodamage. The natural antioxidant improved post-thaw sperm quality, likely by suppressing oxidative stress and apoptosis. Apigenin shows promise for enhancing rooster sperm cryosurvival.

The Prophylactic Effect of Eucheuma spinosum on Total Blood Cholesterol Levels in Rattus norvegicus Dexamethasone-Induced

Hyperlipidemia is one of the main risk factors for heart disease caused by the formation of atherosclerosis. Eucheuma spinosum contains a natural antioxidant flavonoid, which has an antiatherogenic effect that can inhibit the formation of cholesterol in the liver so that it can prevent the development of atherosclerosis. This study aims to determine the effect of prophylactic administration of Eucheuma spinosum on the total cholesterol levels of Rattus novergicus induced by dexamethasone. This study was an experimental design with a post-test-only control group design. This study would use 24 white male Rattus norvegicus divided into three groups; a negative control group (K-) without treatment, a positive control group (K+) induced dexamethasone 8 mg/Kg BW on day 14 until day 20, treatment group (KP) given Eucheuma spinosum extract 200 mg/Kg BW on day 8 until day 20 as Prophylactic and on day 14 until day 20 induced dexamethasone 8 mg/Kg BW. After completing the treatment period, all groups were terminated and taken total calculation of cholesterol levels through the aorta of rats. Data was analyzed statistically computerized. The results of the study of total cholesterol levels that used Kruskal-Wallis test showed differences between groups (p&lt;0.001). There was a decrease in total cholesterol levels in the treatment group (p&lt;0.024) compared with the positive control group. The administration of Eucheuma spinosum extract as a prophylactic dose of 200 mg/Kg BW for 14 days could reduce the total cholesterol level of Rattus norvegicus induced by dexamethasone compared to those not given Eucheuma spinosum extract.

Efficacy of Palmitoylethanolamide and Luteolin Association on Post-Covid Olfactory Dysfunction: A Systematic Review and Meta-Analysis of Clinical Studies.

Post-Covid Olfactory Dysfunction (PCOD) is characterized by olfactory abnormalities, hyposmia, and anosmia, which are among the most often enduring symptoms in individuals who have recovered from SARS-CoV-2 infection. This disorder has been reported to persist in subsets of patients well after 12 months following infection, significantly affecting their quality of life. Despite the high prevalence of PCOD among patients who suffered from SARS-CoV-2 infection, specific therapeutic strategies are still limited. Among these, emerging evidence seems to indicate the administration of CoUltraPEALut, a combination of micronized Palmitoylethanolamide (PEA), an endogenous fatty acid amide, and Luteolin, a natural antioxidant flavonoid, as a viable therapy, especially when given as an adjuvant to olfactory training. Based on the above, a systematic review and a meta-analysis of the literature were conducted, with the aim of evaluating the efficacy of CoUltraPEALut as an addition to olfactory training (OT), in treating PCOD symptoms. Pubmed (MEDLINE), Embase (OVID), and Web of Science scientific databases were screened from the inception until 31 May 2023, and a total of 407 articles were recovered; only five of these studies (441 total patients between treated and control groups) were included in the systematic review. CoUltraPEALut demonstrated significant efficacy in the overall recovery of the olfactory function, compared to the conventional therapy, suggesting that it could represent a possible future adjuvant treatment for PCOD.

Highly Selective and Sensitive Sensor Based IL and CMC-MWCNTs Nanocomposite for Rutin Determination

Rutin is a natural antioxidant flavonoid compound with anti-inflammatory, antioxidant, and antiviral effects that is used to prepare drugs with wide application in clinical treatment. Therefore, the quantitative detection of rutin has important practical significance. In this work, a novel electrochemical sensor based on glassy carbon electrodes (GCEs) modified with sodium carboxymethylcellulose (CMC), multi-walled carbon nanotubes (MWCNTs), and 1-butyl-3-methylimid (ionic liquid, IL) was developed for the super-sensitive detection of the flavonoid rutin. The properties of these modified materials were analyzed by transmission electron microscope (TEM), cyclic voltammograms (CVs), and electrochemical-impedance spectroscopy (EIS). CMC was used to disperse MWCNTs to further enhance their hydrophilicity and biocompatibility. The modified MWCNTs improved the sensitivity of rutin detection. The square-wave voltammetry (SWV) technique showed that the linear range of rutin concentration determination was 0.01 μM to 1 µM and 1 µM to 10 µM. The minimum concentration detection of rutin was 0.83 nM and 6.6 nM, respectively. The proposed sensor presented good selectivity for rutin and successfully analyzed rutin content in the pharmaceutical rutin tablets. These results are consistent with those measured by ultra-high-performance liquid chromatography (UHPLC). Therefore, this sensor has latent application value in the analysis of rutin in food and drug tablets and nutraceutical samples.

Myricetin slows liquid–liquid phase separation of Tau and activates ATG5-dependent autophagy to suppress Tau toxicity

Intraneuronal neurofibrillary tangles composed of Tau aggregates have been widely accepted as an important pathological hallmark of Alzheimer's disease. A current therapeutic avenue for treating Alzheimer's disease is aimed at inhibiting Tau accumulation with small molecules such as natural flavonoids. Liquid–liquid phase separation (LLPS) of Tau can lead to its aggregation, and Tau aggregates can then be degraded by autophagy. However, it is unclear whether natural flavonoids modulate the formation of phase-separated Tau droplets or promote autophagy and Tau clearance. Here, using confocal microscopy and fluorescence recovery after photobleaching assays, we report that a natural antioxidant flavonoid compound myricetin slows LLPS of full-length human Tau, shifting the equilibrium phase boundary to a higher protein concentration. This natural flavonoid also significantly inhibits pathological phosphorylation and abnormal aggregation of Tau in neuronal cells and blocks mitochondrial damage and apoptosis induced by Tau aggregation. Importantly, using coimmunoprecipitation and Western blotting, we show that treatment of cells with myricetin stabilizes the interaction between Tau and autophagy-related protein 5 (ATG5) to promote clearance of phosphorylated Tau to indirectly limit its aggregation. Consistently, this natural flavonoid inhibits mTOR pathway, activates ATG5-dependent Tau autophagy, and almost completely suppresses Tau toxicity in neuronal cells. Collectively, these results demonstrate how LLPS and abnormal aggregation of Tau are inhibited by natural flavonoids, bridging the gap between Tau LLPS and aggregation in neuronal cells, and also establish that myricetin could act as an ATG5-dependent autophagic activator to ameliorate the pathogenesis of Alzheimer's disease.

Nano-Zinc Oxide Induced Pancreatic Toxicity and The Ameliorating Role of Naringenin: Histological and immunohistochemical study

Background: Though Nano-zinc oxide particles (ZnO NPs) are widely applied in biomedicine, bioengineering and cosmetology, a controversy developed between ZnO NPs benefits versus toxicity on biological systems. Naringenin is a natural antioxidant flavonoid. Objective: to explore the histological and immunohistochemical alterations in the rat pancreas following intraperitoneal exposure to two different doses of 35 nm ZnO NPs and to assess the ameliorating role of Naringenin.Materials and Methods: Forty-five adult male rats were split randomly into five groups. Group1 served as control. Groups2 and 4 received a single intraperitoneal injection of 250 and 700 mg/kgbw ZnO NPs. Groups3 and 5 administered ZnO NPs as previously described followed by Naringenin gavaged at a dose of 20mg/kgbw/day once daily for 14 consecutive days. Histological, immunohistochemistry, biochemistry, and morphometry studies were accessed in the pancreatic tissue gained from all animals under the study. Results: Contrasted to the group of control, ZnO NPs exhibited a dose dependent pancreatic tissue and cellular damage manifested as vascular congestion, duct dilatation, fibrosis, and inflammatory cell infiltration. Also, both acinar and B-cells showed degenerating changes varied from just cytoplasmic vacuolization in the ZnO NPs(250mg)-treated rats to severe cell shrinking, pyknosis, cytoplasmic and nuclear fragmentation in the ZnO NPs(700mg)-treated rats. Moreover, ZnO NPs provoked significantly increased mean area percentage of collagen fiber deposition and caspase immunoexpressing, significantly raised fasting blood glucose, serum amylase, lipase and MDA levels besides significant decline regarding insulin immunoexpressing. Naringenin administration induced a great recovery concerning the ZnO NPs (250mg)-treated rats but partial recovery regarding the ZnO NPs (700mg)-treated rats. Conclusion: ZnO NPs potentially persuaded an oxidating stress manifest as structural and functional toxicity in the rat pancreas with great reversibility by Naringenin coadministration. A future work concerning ZnO NPs toxicity on vital organs and Naringenin role in opposing this impact is recommended.

Fabrication, characterization and evaluation of myricetin adsorption onto starch nanoparticles

Myricetin (MY) is a natural antioxidant flavonoid with a variety of biological activities. However, extremely low water solubility, bioavailability, and easy degradation, restrict their application. Recently, increasing interest in starch nanoparticles as a new kind of biocompatible renewable polymer in applications like nanocarriers. This work was to fabricate MY adsorption onto tapioca starch nanoparticles (TSNPs) and evaluate their biological activities. The adsorption mechanism, loading amount, antioxidative capacity, and in vitro release of the loaded MY were also analyzed. The adsorption kinetics and adsorption equilibrium were best explained by a pseudo-second-order model and Freundlich isotherms, respectively. Based on the thermodynamic parameters, adsorption was found to be a spontaneous and exothermic process with a decrease in entropy. MY possessed a maximum equilibrium adsorption capacity of 453 ± 8.07 mg/g. Low cytotoxicity were obtained as described by methylene blue assay, and a sustained release of loaded MY was observed in stimulated gastric (pH 2.0) and intestinal (pH 7.0) fluids. Additionally, the rate of clearance of DPPH free radicals was increased by the adsorption of MY onto TSNPs, which was confirmed by the lower value of 50 % inhibitory concentration (IC50).

Glioma progression is suppressed by Naringenin and APO2L combination therapy via the activation of apoptosis in vitro and in vivo.

Naringenin (NG) is a natural antioxidant flavonoid which is isolated from citrus fruits, and has been reported to inhibit colon cancer proliferation. However, the effects of NG treatment on glioma remain to be elucidated. The present study aimed to explore the effects of NG on glioma in vitro and in vivo. Also, the interactions between NG and APO2 ligand (APO2L; also known as tumor necrosis factor-related apoptosis-inducing ligand) were investigated in glioma. A synergistic effect of NG and APO2L combination on apoptotic induction was observed, though glioma cells were insensitive to APO2L alone. After NG treatment, glioma cells resumed the sensitivity to APO2L and cell apoptosis was induced via the activation of caspases, elevation of decoy receptors 4 and 5 (DR4 and DR5) and induction of p53. Coadministration of NG and APO2L decreased levels of anti-apoptotic B cell lymphoma 2 (Bcl-2) family members Bcl-2 and Bcl-extra large (Bcl-xL), while increased levels of proapoptotic factors Bcl-2-associated agonist of cell death (Bad) and Bcl-2 antagonist/killer 1 (Bak). Furthermore, an in vivo mouse xenograft model demonstrated that NG and APO2L cotreatment markedly suppressed glioma growth by activating apoptosis in tumor tissues when compared with NG or APO2L monotherapy. The present study provides a novel therapeutic strategy for glioma by potentiating APO2L-induced apoptosis via the combination with NG in glioma tumor cells.

Naringenin-Induced Apoptotic Cell Death in Prostate Cancer Cells Is Mediated via the PI3K/AKT and MAPK Signaling Pathways.

Prostate cancer is the most common cancer in men and the second most common cause of cancer-related deaths in men. Although, various drugs targeting the androgen receptor are normally used, the patients frequently undergo recurrence of the disease. To overcome these limitations, natural compounds have been researched for evidence that they suppress progression and metastasis of various cancer cells. In the present study, we investigated effects of naringenin, a natural anti-oxidant flavonoid derived from citrus, on prostate cancer cells (PC3 and LNCaP). Results of present study with PC3 and LNCaP cells revealed that naringenin inhibited proliferation and migration, while inducing apoptosis and ROS production by those cells. In addition, naringenin-induced loss of mitochondrial membrane potential and increased Bax and decreased Bcl-2 proteins in PC3 cells, but not LNCaP cells. In a dose-dependent manner, naringenin decreased phosphorylation of ERK1/2, P70S6K, S6, and P38 in PC3 cells, and reduced phosphorylation of ERK1/2, P53, P38, and JNK proteins in LNCaP cells. However, naringenin activated phosphorylation of AKT in both PC3 and LNCaP cells. Then, targeted signaling proteins associated with viability of PC3 and LNCaP cells were analyzed using pharmacological inhibitors of AKT and ERK1/2 cell signaling pathways. Moreover, we compared the apoptotic effects of naringenin and paclitaxel alone and in combination to find that naringenin enhanced the efficiency of paclitaxel to suppress progression of prostate cancer cell lines. Collectively, these results indicate that naringenin is a potential chemotherapeutic agent for treatment of prostate cancer. J. Cell. Biochem. 118: 1118-1131, 2017. © 2016 Wiley Periodicals, Inc.

Antioxidant, anticancer activities and mechanistic studies of the flavone glycoside diosmin and its oxidovanadium(IV) complex. Interactions with bovine serum albumin

The natural antioxidant flavonoid diosmin, found in citric fruits, showed low antioxidant properties among other flavonoids due to its structural characteristics and low cytotoxicity against lung (A549) and breast (T47D, SKBR3 and MDAMB231) cancer cell lines. The anticancer behavior has been improved by the metal complex generated with the flavonoid and the oxidovanadium(IV) ion. This new complex, [VO(dios)(OH)3]Na5·6H2O (VOdios), has been synthesized and characterized both in solid and solution states. The interaction of the metal ion through the sugar moiety of diosmin precluded the improvement of the antioxidant effects. However, the cell-killing effects tested in human lung A549 and breast T47D, SKBR3 and MDAMB231 cancer cell lines, were enhanced by complexation. The anti-proliferative effects on the human lung cancer cell line were accompanied by cellular ROS generation and an increase in cytoplasm condensation. The breast cancer cell lines did not produce caspase3/7 activation, mitochondrial potential reduction and ROS generation. Therefore, a non-apoptotic form of cell death in a caspase- and oxidative stress-independent manner has been proposed. The protein binding ability has been monitored by the quenching of tryptophan emission in the presence of the compounds using bovine serum albumin (BSA) as a model protein. Both compounds could be distributed and transported in vivo and the complex displayed stronger binding affinity and higher contributions to the hydrogen bond and van der Waals forces.

Formation and inhibition of genotoxic malonaldehyde from DNA oxidation controlled with EDTA

The attack of DNA by reactive oxygen species, such as hydroxyl radical, causes several types of damage, which subsequently promote diseases. Determination of oxidized products, such as malonaldehyde (MA), from DNA would provide theoretical and practical information on the mechanisms of DNA oxidation following DNA damage and this information could be used to prevent DNA damage caused by oxidation. In the present study, calf thymus DNA was oxidized by Fenton’s reagent/EDTA with or without natural antioxidants—flavonoids and anthocyanins—and synthetic antioxidants, Trolox ® and 2H-pyrrole, 3,4-dihydro-2,2-dimethyl-, 1-oxide (DMPO). Amounts of MA formed, which was determined by gas chromatography, in oxidized DNA with the presence of antioxidants, ranged from 7.35 ± 0.88 nmol/mg (2″-O-GIV) to 12.6 ± 0.24 nmol/g (cyanidin). Except for cyanidin, all antioxidants tested inhibited MA formation. DMPO and Trolox ® inhibited MA formation by 12.4% and 27.3%, respectively from oxidized DNA. The decreasing order of inhibitory effect by the anthocyanins was callistephin (30.2%) > keracyanin (27.3%) > Pelargonindin (10.1%) > cyanidin (0%). The decreasing order of inhibitory effect by the flavonoids was 2″-O-GIV (42.7%) > catechin (8.8%) > quercetin (36.4%) > apigenin (34.4%). It is hypothesized that EDTA controlled formation of hydroxyl radicals via trapping Fe(II) ions reversibly.

Alterations in Regional Brain Neurotransmitters by Silymarin, a Natural Antioxidant Flavonoid Mixture, in BALB/c Mice

Silymarin, a natural antioxidant flavonoid mixture, exerts anti-inflammatory effects in the liver and hinders tumor formation. The effect of this flavonoid mixture on the central nervous system is unknown, although antioxidants are considered beneficial. Brain amines and metabolites were studied after a short-term silymarin treatment. BALB/c mice were intraperitoneally treated with 0, 10, 50, or 250 mg/kg of silymarin per day for 5 days. High-performance liquid chromatography coupled with electrochemical detection was performed to determine concentrations of norepinephrine (NE), dopamine (DA), dioxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxytryptamine (5-HT, serotonin) and 5-hydroxyindoleacetic acid (5-HIAA) in discrete brain regions. Analyses showed increased 5-HT levels in the cortex and increased DA and NE levels in the cerebellum in the highest dose group. Results indicated lack of general adverse effect on the brain amine metabolism and suggest that silymarin may have marginal serotonergic, dopaminergic, and noradrenergic effects.

Physiological responses to a natural antioxidant flavonoid mixture, silymarin, in BALB/c mice: II. alterations in thymic differentiation correlate with changes in c-myc gene expression.

Silymarin is a mixture of bioactive flavonoids isolated from the seeds and fruits of Milk Thistle [ Silybum marianum (L.) Gaertner]. We tested the hypothesis that exposure to silymarin will modulate differentiation and cell selection in the thymus via alterations in gene expression. Male BALB/c mice were treated intraperitoneally once daily for five days with 0, 10, 50 or 250 mg/kg of silymarin. Flow cytometric examination of thymic lymphocyte populations showed that the absolute numbers of CD4 + and CD8 + positive T-lymphocytes were increased by silymarin. The c- myc proto-oncogene is important in controlling differentiation and functions of thymocytes. Treatment with silymarin resulted in increased c- myc expression in the thymus. In contrast, the expressions of IL-2 and IL-4 were decreased by silymarin, while MHC II expression did not change. These results indicate that in vivo exposure to silymarin influences phenotypic selection processes in the thymus at doses that may be encountered in natural medicinal use. Further studies investigating the effects of silymarin on the immune system are warranted.

Physiological responses to a natural antioxidant flavonoid mixture, silymarin, in BALB/c mice: I induction of transforming growth factor beta1 and c-myc in liver with marginal effects on other genes.

Silymarin, a mixture of flavonolignans isolated from Silybum marianum, is known for its hepatoprotective properties. We investigated the expression of cytokines in mouse liver following treatment with 0, 10, 50, and 250 mg/kg of silymarin once daily for 5 days. A dose-related but insignificant decrease of circulating alanine aminotransferase and aspartate aminotransferase after silymarin treatment was observed, suggesting that silymarin treatment did not induce hepatic damage. Silymarin treatment caused significant increases in the expressions of transforming growth factor (TGF) beta1 and c-myc in liver. No significant difference was detected among these treatments in the expression of hepatocyte growth factor, interferon gamma, tumor necrosis factor alpha, and class II major histocompatibility complex. These results suggest that alterations of TGFbeta1 and c-myc expression in the liver may be involved in the hepatoprotective effects of silymarin observed in other studies.