Taxifolin Research Articles

Taxifolin, a novel food, attenuates acute alcohol-induced liver injury in mice through regulating the NF-κB-mediated inflammation and PI3K/Akt signalling pathways

Context Taxifolin (TAX) has effective anti-inflammatory, antioxidant and hepatoprotective activities, but its potential mechanism has not been revealed. Objective To evaluate the potential protective effect of TAX on acute alcohol-induced liver injury in mice. Materials and methods Alcoholic liver injury model was established by oral alcohol in mice, and randomly distributed in five groups (n = 10): Normal group (oral saline only); Alcohol group (concentration of fermented alcohol: 56%, 6 mL/kg); TAX groups, mice were orally administered with alcohol, and then TAX with doses of 20, 40, 80 mg/kg, respectively. Oral administration was conducted for 6 weeks. Results TAX treatment illustrated that the level of alanine aminotransferase (ALT) was reduced to 65.90 ± 2.26 U/L and aspartate aminotransferase (AST) to 33.28 ± 5.62 U/L compared with alcohol group (ALT 124.51 ± 4.40 U/L, AST 61.70 ± 4.09 U/L), while superoxide dismutase (SOD) was increased to 49.81 ± 2.39 U/mg and glutathione (GSH) to 8.16 ± 0.44 μmol/g, but MDA was reversed to 2.53 ± 0.24 nmol/mg. Histopathological examination showed TAX treatment alleviated alcohol-induced hepatocyte necrosis and inflammatory infiltration. Meanwhile, Western blot and rt-PCR indicated TAX reduced IL-6 to 2.49 ± 0.25 pg/mL and TNF-α to 1.79 ± 0.20 pg/mL, and inhibiting NF-κB activation in liver. Moreover, TAX reversed alcohol-induced apoptosis by regulating the expression of PI3K/Akt and its downstream apoptotic factors. Conclusions The research provides novel evidence of the hepatoprotective effect of TAX on alcohol-induced liver injury, while also providing the possibility for future treatment of alcoholic liver disease.

Dihydroquercetin influence on clinical and biochemical blood parameters of pigs under conditions of stress load

In modern society, distress has become a widespread condition that negatively affects the functioning of all systems of the human organism. The study of biological mechanisms and changes in the organism under the influence of stress, as well as methods of their leveling, are relevant in medicine, animal science and veterinary medicine. Pigs are an excellent biological model that is closest to humans. The aim of the research was to study the hematological and biochemical parameters of pigs out of and under stress, including against the background of daily consumption of the flavonoid dihydroquercetin (DHQ) with feed. Material and methods. The research was conducted in the experimental yard of the L.K. Ernst Federal Science Center for Animal Husbandry on 3 groups of pigs [F2 hybrid (large white×Landrace)×Duroc] with an initial body weight of 30-35 kg (n=27). Group 1K consisted of control animals not exposed to stress (n=9); group 2K - control animals subjected to simulated stress by the rearrangement of animals (n=9); group 3O - experimental animals subjected to simulated stress and fed throughout the entire experiment DHQ (32 mg per 1 kg of feed) (n=9). On days 0, 42, and 76, blood was collected from the animals and their hematological and biochemical parameters were studied using conventional methods. Results. The positive effect of using DHQ in pigs' nutrition on enhancing the oxidizing function of blood, metabolic intensity, and increasing the endurance of animals under stress conditions has been manifested in maintaining leukocyte level with a higher content of erythrocytes and hematocrit. In animals fed DHQ, alanine aminotransferase activity was lower than in animals not receiving DHQ. Stress led to a significant increase in lactate dehydrogenase activity in group 2K on the 46th day, which was not observed in animals treated with DHQ. Conclusion. Long-term intake DHQ (up to 72 days inclusive) against the background of stress contributed to the preservation of blood values at the control level (without stress), within the physiological norm.

In vitro effects of nutraceutical treatment on human osteoarthritic chondrocytes of females of different age and weight groups.

The in vitro effects of four nutraceuticals, catechin hydrate, gallic acid, α-tocopherol and ascorbic acid, on the ability of human osteoarthritic chondrocytes of two female obese groups to form articular cartilage (AC) tissues and to reduce inflammation were investigated. Group 1 represented thirteen females in the 50-69 years old range, an average weight of 100 kg and an average body mass index (BMI) of 34⋅06 kg/m2. Group 2 was constituted of three females in the 70-80 years old range, an average weight of 75 kg and an average BMI of 31⋅43 kg/m2. The efficacy of nutraceuticals was assessed in monolayer cultures using histological, colorimetric and mRNA gene expression analyses. AC engineered tissues of group 1 produced less total collagen and COL2A1 (38-fold), and higher COL10A1 (2⋅7-fold), MMP13 (50-fold) and NOS2 (15-fold) mRNA levels than those of group 2. In comparison, engineered tissues of group 1 had a significant decrease in NO levels from day 1 to day 21 (2⋅6-fold), as well as higher mRNA levels of FOXO1 (2-fold) and TNFAIP6 (16-fold) compared to group 2. Catechin hydrate decreased NO levels significantly in group 1 (1⋅5-fold) while increasing NO levels significantly in group 2 (3⋅8-fold). No differences from the negative control were observed in the presence of other nutraceuticals for either group. In conclusion, engineered tissues of the younger but heavier patients responded better to nutraceuticals than those from the older but leaner study participants. Finally, cells of group 2 formed better AC tissues with less inflammation and better extracellular matrix than cells of group 1.

Phytochemicals Analysis and Some Bioactive Properties of Erica manipuliflora Salisb. (EMS); Antibacterial, Antiradical and Anti-lipid Peroxidation.

In the present study, the phenolic content by using LC MS/MS method, anticholinergic, antioxidant (metal reduction, radical and removal of lipid peroxidation), and antibacterial activities of Erica manipuliflora Salisb. (EMS) extract were determined. The amount of vanillic acid, fumaric acid, catechin hydrate, quercetin, and phloridzin dihydrate were found higher than other compounds. The ethanol extract of the EMS showed an inhibition effect on the Acetylcholinesterase (AChE) enzyme with IC50 value of 0.124 ± 0.008 mg mL-1. Also, this extract of the EMS indicated radical (DPPH and ABTS) scavenging activity (about 20%) and the reducing capacity for Cu(II) and Fe(III) close to trolox, and inhibited the oxidation of linoleic acid with 40.5% at 20 μg mL-1. Antibacterial activity of the extracts was investigated against Staphylococcus aureus, Escherichia coli, and Salmonella Typhimurium using agar disc diffusion and minimum inhibitory concentration (MIC) methods. The EMS extract was found to be effective when used at least 312 mg mL-1 concentration on the pathogenic microorganisms. Consequently, it has an important non-synthetic natural content that can be used in the treatment of many diseases due to its many bioactivities such as anticholinergic, antioxidant (radical removal, lipid peroxidation prevention, etc.) and antibacterial.

The interaction of the green tea polyphenol (catechin) with pepsin: Insights from spectroscopic to molecular dynamics studies

Pepsin is an aspartic proteinase that plays an essential role in controlling many biological processes, therapy, and pharmaceutical research. Catechins belong to the group of polyphenols that are the most basic chemical composition of green tea. If catechin gets into the human stomach, the binding targets may be the digestive protease. This paper focuses on the behavior of the pepsin enzyme onto the green tea polyphenol, (+)-catechin hydrate. The effects of catechin hydrate on the structure and activity of pepsin were done utilizing ultraviolet−visible (UV–Vis) spectroscopy, intrinsic fluorescence spectroscopy, circular dichroism (CD), thermal stability, kinetic techniques, as well as molecular docking, and Molecular dynamic simulations (MD) technique. The experimental results from fluorescence spectroscopy revealed that the alterations in pepsin's tertiary structure were caused by catechin hydrate binding. The apparent binding constant Ka, the number of binding sites, and thermodynamic parameters were calculated at three different temperatures. Thermodynamic results revealed that catechin hydrate interacts with pepsin spontaneously by hydrophobic forces. The result of the circular dichroism spectral suggests the secondary structural changes. An increase in the content of the β-turn structure was shown. Kinetic parameters revealed that catechin hydrate inhibited the activity of pepsin. The Molecular dynamic (MD) and docking simulations supported experimental findings. As a result, catechin hydrate could be considered as an inhibitor with adverse effects on pepsin structure and function.

Investigation of Anti-Proliferative Effects of Natural Products Quercetin Hydrate and Catechin Hydrate on Leukemia Lymphocytes

The major problem of conventional cancer therapy is lack of selectivity and induction of harmful side-effects on normal (healthy) cells and tissues. In the recent years, scientific efforts are focused to find a proper approach for highly selective influence on cell viability, as well as induction of cell death in cancer cells only. In this regard, natural herbal products are of great interest due to their low cytotoxicity to normal cells and tissues and their potential as supplements to conventional chemotherapeutics. It is well known that flavonoids exhibit various biological activities, such as anti oxidative, anti bacterial, anti inflammatory, anti viral and anti cancer, and may play a role in cancer prevention. In the present study, the effects of low concentrations of quercetin hydrate and catechin hydrate on cell viability of leukemia lymphocytes were investigated, in order to provide an experimental basis for their future incorporation into newly-synthesized biocompatible nano-formulations.

A Study and Optimization of Complexation of Cobalt Ions with Dihydroquercetin in Aqueous Solutions

This work continues a series of studies devoted to complex formation of ions of biogenic metals with the flavonoid dihydroquercetin (DHQ). The interaction of Со2+ ions with DHQ in aqueous solutions has been investigated. It has been found that, at different pH of a solution, complex compounds (CC) with different stoichiometry are formed; a variation of the pH value of a solution from 6.0 to 7.0 results in the formation of compounds (1)–(3) with the metal : flavonoid ligand ratio (Met : L) from 1 : 2 at рН 6.0 (1), through 2 : 3 at pH 6.4–6.7 (2), to 1 : 1 at рН 6.8–7.0 (3). By using the thermogravimetric method and the data of the elemental analysis, the most probable composition of the compounds with the determination of the amount of bound water has been proposed: [CoL2(H2O)4] for (1), [Co2L3(ОН)(H2O)4] for (2), and [CoL(ОН)(H2O)2] for (3). Conditions for the optimization of product yield in the complexation reaction of Со2+ ions with DHQ in an aqueous solution have been determined for compound (2): the рН value of solution 6.7; the reaction time 15 min; the temperature of the reaction solution 90°С; the molar ratio of the initial reagents DHQ : Со2+ 1 : 1.5; the initial concentration of DHQ 0.020 mol/L and that of Со2+ 0.030 mol/L; and the use of CoSO4 ⋅ 7H2O as a source of cobalt ions. The yield of the product is 81.8%.

INVESTIGATION AND OPTIMIZATION OF COMPLEXATION OF MANGANESE (II) IONS WITH DIHY-DROQUERCETIN IN AQUEOUS SOLUTIONS

The purpose of this work was to study and optimize the complexation reaction of Mn2+ ions with flavonoid dihydroquercetin (DHQ) in aqueous solutions. It was found that the interaction of DHQ (0.015–0.030 M) with manganese (II) salts at molar ratios of DHQ : Mn2+ from 1 : 1 to 1 : 3 at pH 6.9–7.9 leads to the formation of a complex compound (CC) with a stoichiometric ratio of metal : flavonoid ligand (Met : L) 1 : 1. The conditions for the complexation of Mn2+ ions with dihydroquercetin at laboratory conditions optimized for the yield of the product, are proposed. The maximum yield of the complex (95%) was achieved at the interaction of DHQ with Mn(CH3COO)2∙4H2O under conditions: pH 7.7, 70 °C, the reaction time was 15 min, the molar ratio of the initial reagents DHQ: Mn2+ was 1 : 1.5, the initial concentrations were 0.020 M DHQ and 0.030 M Mn2+. The most probable composition of the complex with the determination of the amount of bound water [MnL(OH)(H2O)2] and the structure of the complex were established using the data of thermogravimetry (TG), differential scanning calorimetry (DSC), elemental analysis and X-ray fluorescence analysis (XRF), confirmed by the data of total reflection X-ray fluorescence analysis (TXRF), and EPR spectroscopy data.

Dibutyl phthalate-induced oxidative stress, inflammation and apoptosis in grass carp hepatocytes and the therapeutic use of taxifolin

The widespread use of plastic products has led to the widespread presence of plasticizers in the environment. As a common environmental pollutant, research on plasticizer toxicity is insufficient in fish cells. In particular, research on the toxicity of dibutyl phthalate (DBP) in grass carp hepatocyte lines is insufficient. To further explore these mechanisms, we treated grass carp hepatocytes with 300 μM DBP, a common plasticizer, for 24 h, and hepatocytes were also treated with 1 μM taxifolin (TAX), an antioxidant, for 24 h to study its antagonistic effect on DBP. After DBP exposure, oxidative stress levels and inflammation in hepatocytes increased, and the mRNA and protein expression of apoptosis-related markers increased significantly, leading to hepatocyte apoptosis. Moreover, AO/EB staining, Hoechst staining and flow cytometry also showed that the level of apoptotic cells increased after DBP exposure. Notably, both TAX pretreatment and TAX simultaneous treatment alleviated oxidative stress, increased inflammatory factor levels and apoptosis induced by DBP. In comparison, the effect of simultaneous TAX treatment was better than that of TAX pretreatment. Our results showed that TAX alleviates DBP-induced apoptosis in grass carp hepatocytes through oxidative stress and inflammation, and TAX pretreatment and simultaneous treatment exhibited specific effects. Specifically, simultaneous treatment had a better effect. Our study assessed the toxicity of DBP in grass carp hepatocytes and provided a theoretical and research basis for the in vivo study of animal models in the future. The innovation of this study involves the exploration of the interaction between DBP and TAX for the first time. This study may enrich knowledge regarding the theoretical mechanism of DBP toxicity in fish hepatocytes and propose methods address DBP toxicity.

Determination and Evaluation of Flavonoids in Hawthorn in China

Hawthorn (Crataegus pinnatifida Bunge) is a common agricultural product with applications in medicine and food in China. In particular, the flavonoids in hawthorn have important medicinal effects. To quantify the flavonoid contents in hawthorn fruit of different regions in China and scientifically guide the reasonable consumption of hawthorn, 12 flavonoids in hawthorn samples from Shandong (SD), Henan (HN), Shaanxi (SAX), Shanxi (SX), and Jiangsu (JS) were detected by means of high performance liquid chromatography (HPLC). The total flavonoid content in the Henan sample from hawthorn was the highest, at 177.94 μg · g−1, and the Jiangsu sample was the lowest at 124.67 μg · g−1. Epicatechin was the flavonoid with the highest concentration in samples from all regions, and the average content in Shandong samples reached 73.44 μg · g−1. Principal component analysis (PCA), hierarchical cluster analysis (HCA) and discriminant analysis (DA) were also performed to evaluate the effect of different growing regions in China on the contents of flavonoids in hawthorn. We found the Jiangsu samples had obvious regional characteristics, and the compositions of (±)-catechin hydrate, epicatechin and phlorizin dihydrate could be used to distinguish hawthorn grown in Jiangsu from the other four producing areas. The results will provide the reference basis for Chinese hawthorn consumers.

Feeding dihydroquercetin and vitamin E to broiler chickens reared at standard and high ambient temperatures

ABSTRACT The use of natural antioxidants, in particular polyphenols such as dihydroquercetin (DHQ), in animal nutrition has recently increased in popularity. This may partly be due to the risk of increased incidences of heat stress associated with raising livestock in warmer ambient temperatures, facilitated by global warming, reducing antioxidant capacity. The current research demonstrates the effect of dietary DHQ, vitaminEand standard or high ambient temperatures on growth performance, energy and nutrient metabolism, gastrointestinal tract (GIT) development, jejunal villus morphometry and antioxidant status in broiler chickens. Each of the four experimental diets was fed to 16 pens of five birds, which were allocated to four rooms (four pens in each room). The temperature in two rooms was maintained at aconstant 35°C (high temperature; HT), and the temperature in the other two rooms was gradually reduced from 27°C at 7 dof age to 22°C at 20 dof age (standard temperature; ST). Rearing birds at HT reduced feed intake, weight gain, weight of small intestine, total GIT, liver, spleen, heart, villus height, villus surface area and lowered blood glutationperoxidase (GSH-Px). Dietary DHQ increased blood GSH-Px and total antioxidant status, increased heart weight and reduced caecal size. When fed separately, DHQ and vitamin E improved hepatic vitamin E concentration. Feeding vitamin Eincreased spleen and liver weights. When fed together, DHQ and vitamin Ereduced villus height, villus height to crypt depth ratio and villus surface area. Temperature and antioxidants did not affect energy and nutrient metabolism. There were no effects of dietary antioxidants on growth performance of broiler chickens and there were no mortalities. At present, it is unclear if feeding antioxidants (in particular DHQ) at different levels, using different dietary formulations, and rearing birds under arange of environmental conditions may be effective at enhancing production performance and bird health in hot ambient climates.

Влияние дигидрокверцетина на артериальное давление и гемореологические параметры крыс SHR в периоды развития и стабильной артериальной гипертензии

Согласно современным представлениям, важную роль в прогрессировании артериальной гипертензии (АГ) играет изменение реологических свойств крови. К настоящему времени опубликованы работы, в которых показано улучшение гемореологических параметров за счет использования различных лекарственных средств при сердечно-сосудистых заболеваниях. Цель работы - исследование влияния дигидрокверцетина (ДГК) на артериальное давление (АД) и гемореологические параметры у крыс линии SHR в различные периоды развития АГ. Методика. Эксперименты проведены на крысах со спонтанной гипертензией (spontaneously hypertensive rats, SHR) двух возрастных групп: период развития АГ (возраст крыс SHR на начало эксперимента 5 нед) и период стабильной АГ (возраст крыс SHR на начало эксперимента 11 нед). ДГК (50 мг/кг) вводили внутрижелудочно в течение 6 нед. Крысы контрольных групп получали эквиобъемное количество крахмальной слизи по той же схеме. АД у бодрствующих крыс регистрировали неинвазивным методом (NIBP200A, «Biopac Systems, Inc.», США). Вязкость цельной крови, стабилизированной ЭДТА (в диапазоне скоростей сдвига 15-450 с-1), и плазмы (450 с-1) измеряли на ротационном вискозиметре (LVDV-II+ Pro, CP40, «Brookfield Engineering Labs Inc.») при 36 оС. Гематокрит определяли методом центрифугирования. Агрегацию и деформируемость эритроцитов исследовали на анализаторе «RheoScan-AnD 300» («Rheo Meditech, Inc.»). Результаты. У крыс SHR после 6-недельного введения ДГК в период развития АГ не наблюдалось значимых различий уровня систолического АД (САД), макро- и микрореологических показателей по сравнению с параметрами в контрольной группе. У крыс SHR со стабильной АГ введение ДГК приводило к снижению САД, ослаблению агрегации эритроцитов с увеличением их индекса элонгации и снижению вязкости крови по сравнению с контрольной группой. Заключение. Нарушение гемореологических параметров крови выражено значимо меньше у крыс SHR в начальной стадии формирования АГ. Использование ДГК в период развития АГ не оказывало статистически значимого влияния на АД. В период стабильной АГ гипотензивное действие ДГК связано в том числе с его способностью снижать вязкость крови и улучшать такие микрореологические параметры, как агрегация и деформируемость. Arterial hypertension (AH) is the most common cardiovascular disease and a risk factor for other vascular diseases and organ damage. According to current concepts, changes in blood rheology play an important role in AH progression. Drugs improving hemorheological parameters have been shown to be beneficial in cardiovascular diseases. Aim. To study the effect of dihydroquercetin (DHQ) on blood pressure (BP) and hemorheological parameters in spontaneously hypertensive rats (SHR) at different stages of AH. Methods. Experiments were performed on SHR in the period of AH development (5-week-old SHR at the beginning of experiments) and in the period of stable AH (11-week-old SHR at the beginning of experiments). DHQ (50 mg/kg) was administered intragastrically for 6 weeks. Rats of the control groups received an equal volume of starch mucilage according to the same schedule. BP was measured with a non-invasive NIBP200A (Biopac Systems, Inc., USA) instrument in conscious rats. Viscosity of K2EDTA-stabilized whole blood (shear rate range, 15-450 s-1) and plasma (450 s-1) was measured with a rotational viscometer (LVDV-II + Pro, CP40, Brookfield Engineering Labs Inc.) at 36 °C. Hematocrit was determined by centrifugation. Aggregation and deformability of red blood cells was studied with a RheoScan-AnD 300 analyzer (RheoMeditech, Inc.). Results. After 6 weeks of DHQ administration in the period of AH development, mean blood pressure (MBP) and macro- and microrheological parameters did not significantly differ from the respective values in the control group In SHR with stable AH, the DHQ treatment resulted in decreased MBP, blood viscosity, and red blood cell aggregation and increased red blood cell elongation index compared to the control group. Conclusion. Disorders of hemorheological parameters were significantly less pronounced in SHR with early AH. Perhaps for this reason, the DHQ treatment during the development of hypertension did not exert a significant effect on BP. In the period of stable hypertension, hyperviscosity syndrome progressed in SHR. The antihypertensive effect of DHQ in this period, among other things, was associated with a beneficial effect on micro-rheological parameters, including red blood cell aggregation and deformability, which was evident as decreased blood viscosity.

Effect of modelled stress and adaptogens on microstructural characteristics of pork from fast-growing hybrid animals

This research aimed to study the effect of modelled technological stress and the introduction of selenium and dihydroquercetin (DHQ) into pig diets on the microstructure of M. longissimus dorsi muscle tissue. The in vivo experiment was carried out on 36 hybrid young barrows (Large White x Landrace) x Duroc) with an initial live weight of 34 – 36 kg until they reached a weight of not less than 110 kg. The animals were divided into four groups: 1 (C-) – pigs did not receive adaptogens and were not subjected to modelled technological stress; 2 (C+) – pigs did not receive adaptogens but were subjected to stress via relocation of animals; 3 (С+Se) – pigs were subjected to stress and received 0.2 mg Se.kg-1 feed as selenium proteinate in addition to their diet; 4 (С+DHQ) – pigs were subjected to stress and received 32 mg DHQ.kg-1 feed in addition to their diet. The best results regarding the muscle tissue condition were recorded in the musle L. dorsi samples were taken from the carcasses of group 4 (С+DHQ). Analysis of variance using the Fisher–Snedecor test confirmed that addition of adaptogens led to an improvement of the pH24 value (at p = 0.05, f observed = 5.90 >fcritical = 4.17) and moisture-holding capacity (at p = 0.05, f observed = 3.04 >fcritical = 2.92). The effect of long-term addition of DHQ to pig diets (78 days) on the condition of muscle tissue was studied for the first time, which allowed us to conclude its role in the prevention of myopathic changes in the muscle fibre structure.

Functional characterization of three flavonol synthase genes from Camellia sinensis: Roles in flavonol accumulation

Flavonol derivatives are a group of flavonoids benefiting human health. Their abundant presence in tea is associated with astringent taste. To date, mechanism pertaining to the biosynthesis of flavonols in tea plants remains unknown. In this study, we used bioinformatic analysis mining the tea genome and obtained three cDNAs that were annotated to encode flavonol synthases (FLS). Three cDNAs, namely CsFLSa, b, and c, were heterogenously expressed in E. coli to induce recombinant proteins, which were further used to incubate with three substrates, dihydrokampferol (DHK), dihydroquercetin (DHQ), and dihydromyricetin (DHM). The resulting data showed that three rCsFLSs preferred to catalyze (DHK). Overexpression of each cDNA in tobacco led to the increase of kampferol and the reduction of anthocyanins in flowers. Further metabolic profiling of flavan-3-ols in young tea shoots characterized that kaempferol derivatives were the most abundant, followed by quercetin and then myricetin derivatives. Taken together, these data characterized the key step committed to the biosynthesis of flavonols in tea leaves. Moreover, these data enhance understanding the metabolic accumulation relevance between flavonols and other main flavonoids such as flavan-3-ols in tea leaves.

Prospects for the use of regulators of oxidative stress in the comprehensive treatment of the novel Coronavirus Disease 2019 (COVID-19) and its complications.

Some surface proteins of the newly identified severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can bind to the hemoglobin molecule of an erythrocyte, which leads to the destruction of the structure of the heme and the release of harmful iron ions to the bloodstream. The degradation of hemoglobin results in the impairment of oxygen-carrying capacity of the blood, and the accumulation of free iron enhances the production of reactive oxygen species. Both events can lead to the development of oxidative stress. In this case, oxidative damage to the lungs leads then to the injuries of all other tissues and organs. The use of uridine, which preserves the structure of pulmonary alveoli and the air-blood barrier of the lungs in the course of experimental severe hypoxia, and dihydroquercetin, an effective free radical scavenger, is promising for the treatment of COVID-19. These drugs can also be used for the recovery of the body after the severe disease.

Pistachio Green Hull Extract Induces Apoptosis through Multiple Signaling Pathways by Causing Oxidative Stress on Colon Cancer Cells.

Pistachio is considered to be one of the fifty foods with the highest antioxidant effect. However, the anticancer effect mechanisms of this plant extracts are unknown. The aim of this study was to investigate the anticancer effect of different extracts from the green hull of pistachio. The cytotoxic effects of different solvent extracts on cancer and normal cells were examined by cell viability assay and flow cytometric analysis. The levels of the apoptotic gene and protein were investigated by Western Blot and ELISA, and qPCR. The intracellular free radical exchange was determined by oxidative and nitric oxide analyses. DNA damage level was measured by the 8-OHdG test. Phenolic and free fatty acid components were examined by LC-MS/MS and GC-MS, respectively. It was determined that the n-hexane fraction showed a higher cytotoxic effect on cancer cells. Oxidative and cell cycle analyses indicated that the n-hexane fraction arrested cell cycle of HT-29 at the sub-G1 phase by increasing DNA damage through oxidative stress. In addition, gene expression analysis of the HT-29 treated with the n-hexane fraction indicated that apoptotic and autophagic gene expressions were significantly upregulated. LC-MS/MS analysis of the n-hexane fraction revealed the presence of 15 phenolic compounds, containing mainly gallic acid and catechin hydrate, and GC-MS analysis determined the presence of the following fatty acids: 9-octadecenoic acid, 9,12-octadecadienoic acid and hexadecenoic acid. Based on these grounds, we suggest that the n-hexane fraction of pistachio green hull damages DNA, arrests the cell cycle at the G1 subphase, and induces apoptosis through oxidative pathways in colon cancer.

Assessment of preclinical effect of (+)-catechin hydrate on sexual function: An in silico and in vivo study.

Considering dopamine-enhancing effect of (+)-catechin, the present study was designed to evaluate dopamine-2 (D2) receptor agonistic and phosphodiesterase-5 (PDE5) enzyme inhibitory effects in in silico and effect on male sexual function of Sprague Dawley rats in vivo. (+)-Catechin and standard (sildenafil and bromocriptine) were docked using Autodock Vina 1.1.2 and visualised by UCSF Chimera 1.14. Significant interactions in terms of binding energies were observed for catechin with both proteins. In in vivo study, the rats were dosed orally for 54days with (+)-catechin hydrate (50mg/kg), sildenafil citrate (standard, 4mg/kg) and carboxymethylcellulose (vehicle, 0.25%w/v). The aphrodisiac effects were evaluated on the day 14, 28, 42 and 54 using the behavioural parameters of mounting and intromission. After the study, animals were sacrificed and testes and spermatozoa were assessed for safety profile. Results showed a significant increase in mount and intromission frequencies and a significant reduction in mount and intromission latencies in the catechin group on all tested days when compared to vehicle control. (+)-Catechin was found to be safe on histology of testes, sperm count, sperm motility and sperm morphology parameters. In conclusion, catechin demonstrated an enhancement in sexual behaviour without eliciting toxicity on the male reproductive system in rats.

Biosynthesis of phenylpropanoids and their protective effect against heavy metals in nitrogen-fixing black locust (<i>Robinia pseudoacacia</i>)

Purpose: To examine the effect of various heavy metals (HMs) on phenylpropanoid pathway compounds in Robinia pseudoacacia.Methods: A series of pot culture experiments were performed to understand how the metabolic profile of phenylpropanoid compounds were affected by various HMs, such as redox-active HMs (AgNO3 and CuCl2), and non-redox-active HMs (HgCl2). Phenylpropanoid compound level was evaluated by high performance liquid chromatography.Results: The total phenylpropanoid level in leaves increased significantly in all the treated groups when compared to that in the untreated group (p < 0.05). However, a significant effect on the total phenylpropanoid levels was only found for redox-active HMs (p < 0.05), whereas non-redox-active HMs showed less accumulation. Chlorogenic acid and rutin were the two major phenylpropanoid compounds found after the plants were subjected to redox and non-redox-active HMs stress. However, when compared to these two compounds, the levels of catechin hydrate, epicatechin, p-coumaric acid, kaempferol, and quercetin were lower. Caffeic acid level was significantly decreased in both redox and non-redox-active HMs when compared to that in the control (p < 0.05). In addition, trans-cinnamic acid accumulation was altered based on the types and concentration of HMs.Conclusion: Phenylpropanoid metabolic pathway participated in the HM tolerance process for the protection of R. pseudoacacia from oxidative damage caused by HMs, thus allowing the species to grow in highly HMs-contaminated areas.
 Keywords: Heavy metals, Non-redox-active metals, Phenylpropanoid compounds, Redox-active metals, Robinia pseudoacacia

Эндотелиопротекторные свойства ATACL и катехин гидрата в условиях истощающих нагрузок крыс

Цель исследования - изучение эндотелиопротекторных свойств ATACL и катехин гидрата в сравнении с известным лекарственным препаратом Сулодексид в условиях истощающих нагрузок крыс. Методика. Истощающие нагрузки воспроизводили методом Порсолта - принудительного плавания животных с нагрузкой до полного истощения и борьбы за жизнь. Эксперименты выполнены на 50 крысах-самцах Wistar (200-220 г), разделенных на 5 групп (n=10). Вазодилатирующую функцию эндотелия сосудов головного мозга оценивали методом допплерографии с использованием программного комплекса «Minimax Doppler», производства компании «Минимакс» (Санкт-Петербург, Россия). Результаты. Показано, что на фоне истощающих нагрузок наблюдалось значительное ухудшение вазодилатирующей и антитромботической функций эндотелия мозговых сосудов, что проявлялось снижением реакции сосудов в ответ на введение анализаторов (АЦХ и L- NAME), развитием «феномена L-аргининового парадокса», повышением как степени, так и агрегационной способности (скорости) тромбоцитов. Исследуемые соединения ATACL и катехин гидрат способствовали коррекции эндотелиальной дисфункции, что выражалось в статистически значимом увеличении скорости кровотока при внутривенном введении АЦХ у животных групп, получавших ATACL на 26,1%, катехин гидрат - на 24,5%, относительно исходной скорости кровотока. ATACL (4-гидрокси-3,5-ди-трет-бутилкоричнаякислота) оказывал выраженное влияние на антитромботическую функцию, при этом статистически значимо снижалась степень агрегации тромбоцитов на 99,3% и скорости агрегации - на 136,6%, относительно аналогичных показателей группы крыс негативного контроля. Заключение. Соединение ATACL представляет интерес для дальнейшего изучения в качестве актопротекторного средства, обладающего эндотелиопротекторной активностью. Aim. To study possible endothelium-protective properties of catechin hydrate and ATACL in comparison with a well-known drug Sulodexide in exhaustive physical exercise of rats. Methods. Exhaustive exercise was produced by forced swimming of rats with a load until exhaustion and struggle for life. Experiments were performed on 50 Wistar male rats (200-220 g) divided into 5 groups (n=10 in each group). The endothelial vasodilatory function of cerebral blood vessels was evaluated with the Doppler method using Minimax Doppler equipment and software (Minimax, St. Petersburg, Russia). Results. Exhaustive exercise induced a significant impairment of vasodilatory and antithrombotic endothelial functions, which was evident as decreased response of cerebral blood vessels to acetylcholine (ACH) and L-NAME, development of the L-arginine paradox, and increases in both platelet aggregation degree and rate. ATACL (4-hydroxy-3,5-di-tert-butyl cinnamic acid) and catechin hydrate provided correction of the endothelial dysfunction, which was evident from increased blood flow velocity in response to intravenous administration of ACH by 26.1% in the ATACL group (p<0.05) and by 24.5% in the catechin hydrate group (p<0.05) compared to the baseline blood flow velocity. ATACL exerted a pronounced effect on the endothelial antithrombotic function evident as decreases in aggregation degree by 99.3% (p<0.05) and aggregation rate by 136.6% (p<0.05) compared to the group of negative control. Conclusion. ATACL is of interest for further studies as an actoprotector with endothelium-protective activity.

A Chitosan-PLGA based catechin hydrate nanoparticles used in targeting of lungs and cancer treatment.

ObjectiveTo prepare a novel Chitosan (CS)-coated-PLGA-NPs of catechin hydrate (CTH) and to improve lungs bioavailability via direct nose to lungs-delivery for the comparative assessment of a pulmokinetics study by the first-time UHPLC-MS/MS developed method in the treatment of lungs cancer via anticancer activities on H1299 lung cancer cells.Material and methodsPLGA-NPs was prepared by solvent evaporation (double emulsion) method followed by coated with chitosan (CS) and evaluated based on release and permeation of drug, a comparative pulmokinetics study with their anticancer activities on H1299 lung cancer cells.ResultsThe particle size, PDI and ZP of the optimized CAT-PLGA-NPs and CS-CAT-PLGA-NPs were determined 124.64 ± 12.09 nm and 150.81 ± 15.91 nm, 0.163 ± 0.03 and 0.306 ± 0.03, –3.94 ± 0.19 mV and 26.01 ± 1.19 mV respectively. Furthermore, higher entrapment efficiency was observed for CS-CAT PLGA NPs. The release pattern of the CS-CAT-PLGA NPs was found to favor the release of entrapped CAT within the cancer microenvironment. CS-CAT-PLGA-NPs exposed on H1299 cancer cells upto 24.0 h was found to be higher cytotoxic as compared to CAT-solution (CAT-S). CS-CAT-PLGA-NPs showed higher apoptosis of cancer cells after their exposure as compared to CAT-S. CS-CTH-PLGA-NPs showed tremendous mucoadhesive-nature as compared to CTH-S and CS-CTH-PLGA NPs by retention time (RT) of 0.589 min, and m/z of 289.21/109.21 for CTH alongwith RT of 0.613 min and m/z of 301.21/151.21 was found out for IS (internal standard), i.e. Quercetin). Likewise, for 1–1000 ng mL−1 (linear range) of % accuracy (92.01–99.31%) and %CV (inter & intra-day, i.e. 2.14–3.33%) was determined. The improved Cmax with AUC0–24 was observed extremely significant (p < 0.001) via i.n. as compared oral and i.v. in the wistar rat’s lungs. The CS-approach was successfully designed and safely delivered CAT to the lungs without causing any risk.ConclusionCS-CTH-PLGA-NPs were showed a significant role (p < 0.001) for the enhancement of lungs-bioavailability and potentially promising approach to treat lung cancers. CS-CTH-PLGA-NPs did not cause any toxicity, it showed safety and have no obvious toxic-effects on the rat’s lungs and does not produce any mortality followed by no abnormal findings in the treated-rats.