Rat Liver Homogenates Research Articles

Cadmium Chloride-induced skin ulceration and inflammation in rats: Anti-inflammatory potentials of Moringa oleifera and Musa sapientium via NF-KB/IL-4/1L-10-mediated pathway.

Cadmium-induced toxicity resulted in skin carcinogenesis in rats. The mechanism underlying Cadmium-induced skin carcinogenesis remains poorly understood. Therefore, we evaluated anticancer potentials of Moringa oleifera and Musa sapientum in Cadmium Chloride (CdCl2)-induced skin carcinogenesis, ulceration and inflammation in rats. Twenty-eight adult male wistar rats (average weight of 155 g) were randomly divided into 7 groups (n = 4). Group 1 was control. Groups 2-4 and 7 received single intraperitoneal administration of 1.5 mg/Kg bodyweight of CdCl on Day 1. Thereafter, Groups 3, 4 and 7 were post-treated with 15 mg/Kg bodyweight of MO11, 15 mg/Kg bodyweight of MO11 + 7 mg/Kg bodyweight of MS06 and 3.35 mg/Kg bodyweight of Doxorubicin respectively (Days 1-17). Groups 5 and 6 received only MO11-dose and Olive Oil (vehicle) respectively (Days 1-17). Skin histopathology (Heamatoxyline and Eosin technique) and ELISA concentrations of pro-inflammatory biomarkers (IL-1β, IL-6, IL-8 and NF-kB) and antiinflammatory biomarkers (IL-4 and IL-10) in liver homogenates of rats of Groups 1-7 were evaluated. Computed data were statistically analysed using Mann-Whitney-U test at P ≤ 0.05. Histo-pathological evaluations showed normal skin histology in Groups 1 and 3-5, but skin histoalterations in Groups 2 and 6. Post-treatments of CdCl2-induced skin carcinogenesis with MO11, MO11+MS06 and Doxorubicin resulted in downregulations of IL-1β, IL-6, IL-8 and NF-kB, butupregulations of IL-4 and IL-10 in Groups 3, 4 and 6, compared with Group 2. MO11 and MO11+MS06 achieved better anti-inflammatory potentials than Doxorubicin. Overall, MO11 and MS06 possess anticancer and anti-inflammatory potentials.

Metabolism of nitric oxide under the conditions chronic alcohol intoxication modelling

It was experimentally shown that ethanol affects the production of nitric oxide in rats. However, nitric oxide can have both a protective effect by weakening the harmful effect of ethanol on the microcirculation of the liver, and lead to liver damage by active forms of nitrogen. The purpose of the study is to study changes in the nitric oxide cycle under the conditions of modeling chronic alcohol intoxication in rats. Experiments were performed on 30 white, mature male Wistar rats, weighing 180–220 g. The animals were divided into 2 groups: I – control (n=6); II group – animals with alcoholic hepatitis (n=24) modelled by the method of forced intermittent alcoholization for 5 days, with a repeat after two days by intraperitoneal injection of a 16.5% ethanol solution in a 5% glucose solution, at the rate of 4 ml/kg of body weight. Animals were removed from the experiment on days 10, 14, 21 and 28 by taking blood from the right ventricle of the heart under thiopental anesthesia. The activity of inducible and constitutive isoforms of NO-synthase, concentration of nitrite, nitrosothiols and peroxynitrites of alkali and alkaline earth metals, the activity of nitrite reductase, nitrate reductase and arginase were determined in rat liver homogenate. Chronic alcohol intoxication modelling for 10–28 days leads to a violation of the formation and metabolism of nitric oxide with the predominant formation of its toxic metabolites, such as peroxynitrites and nitrites, which threatens the development of nitrosative stress in the liver. Chronic alcohol intoxication on the 10th–28th days of the experiment is accompanied by a sharp decrease in the activity of the arginase-dependent pathway of arginine metabolism in the liver of rats, which indicates a violation of the deamination processes in the Krebs-Handzeleit cycle. Keywords: nitrites, NO-synthase, peroxynitrite, alcohol, liver, rats.

A Comparative Study on GC-MS Profiles and Effect of Ethanolic Extracts of Stem-Bark and Leaves of Daniellia oliveri on Cerebral Sodium Pump and Thiols Status in Fenton Reaction Treated Rat Organs In-vitro

Daniellia oliveri is one of the most extensively utilized medicinal plants in Nigeria and some West African countries for the treatment of various ailments. In the present study, the GC-MS profiles and effect of ethanolic extracts of stem bark and leaves of Daniellia oliveri on lipid peroxidation, Na+/K+-ATPase activity and thiols status in H2O2 and Fe2+ (fenton reaction) assaulted cerebral and hepatic tissue homogenates was investigated. The stem bark and leaves were extracted with absolute ethanol for 72hrs and concentrated in a rotary evaporator. Wistar rats were euthanized and the brains and livers were removed, homogenized, centrifuged and the supernatants were used for lipid peroxidation, Na+/K+-ATPase activity, and thiol assays in the presence of prooxidants (1mM H2O2 or 10μM Fe2+ or H2O2 and Fe2+).The results revealed that ethanol extract of both the leaf and stem-bark inhibited lipid peroxidation induced by H2O2, Fe2+ and combination of H2O2 and Fe2+ and this was evident in the reduction of lipid peroxidation adducts formation in rat liver and brain homogenates with the stem-bark possessing more efficacy. Furthermore, the results of Na+/K+-ATPase and thiol assays revealed that H2O2 and Fe2+ inhibited the activity of cerebral Na+/K+-ATPase, while H2O2/Fe2+ caused a marked reduction in the levels of both protein and non-protein thiols in the tissues homogenates. However, the ethanolic extract of Daniellia oliveri extirpated this anomaly by increasing the activity of cerebral Na+/K+-ATPase as well as the thiol level in the rat hepatic and cerebral tissues. Finally, GC-MS analysis was carried out on the ethanolic extracts of the stem bark and leaves of Daniellia oliveri and results revealed the presence of 21 compounds including 1,2-Benzisothiazol-3-amine, 1H-Indole-2-carboxylic acid, 2-Ethylacridine, Ethyl 2-(2-chloroacetamido)-3,3,3-trifluoro-2-(3-fluoroanilino) propionate and trans-1-Butyl-2-methylcyclopropane in the stem bark and 10 compounds including Methylthio-acetonitrile, 2-Methyl-Z,Z-3,13-octadecadienol, Desulphosinigrin and 4H-1,2,4-triazole-3,5-diamine in the leaves, justifying the observed biological activities of Daniellia oliveri and the higher potency of the stem bark may be attributed to the presence of more bioactive constituents found in it. This study therefore justifies the medicinal usage of leaves and stem-bark of Daniellia oliveri and suggests its consideration in the treatment and management of degeneration diseases with etiology associated with oxidative stress.

Evaluation of Selenomethionine Entrapped in Nanoparticles for Oral Supplementation Using In Vitro, Ex Vivo and In Vivo Models

Selenium methionine (SeMet) is an essential micronutrient required for normal body function and is associated with additional health benefits. However, oral administration of SeMet can be challenging due to its purported narrow therapeutic index, low oral bioavailability, and high susceptibility to oxidation. To address these issues, SeMet was entrapped in zein-coated nanoparticles made from chitosan using an ionic gelation formulation. The high stability of both the SeMet and selenomethionine nanoparticles (SeMet-NPs) was established using cultured human intestinal and liver epithelial cells, rat liver homogenates, and rat intestinal homogenates and lumen washes. Minimal cytotoxicity to Caco-2 and HepG2 cells was observed for SeMet and SeMet-NPs. Antioxidant properties of SeMet were revealed using a Reactive Oxygen Species (ROS) assay, based on the observation of a concentration-dependent reduction in the build-up of peroxides, hydroxides and hydroxyl radicals in Caco-2 cells exposed to SeMet (6.25-100 μM). The basal apparent permeability coefficient (Papp) of SeMet across isolated rat jejunal mucosae mounted in Ussing chambers was low, but the Papp was increased when presented in NP. SeMet had minimal effects on the electrogenic ion secretion of rat jejunal and colonic mucosae in Ussing chambers. Intra-jejunal injections of SeMet-NPs to rats yielded increased plasma levels of SeMet after 3 h for the SeMet-NPs compared to free SeMet. Overall, there is potential to further develop SeMet-NPs for oral supplementation due to the increased intestinal permeability, versus free SeMet, and the low potential for toxicity.

Gender determinism of the effect of placenta cryoextract on the hepatotropic effects of esomeprazole, clarithromycin and metronidazole in chronic liver damage

Current regimens for the eradication of Helicobacter pylori infection include bismuth drugs, proton pump inhibitors, and a combination of 2–3 antibacterial drugs. Uncontrolled use of antibacterial agents as part of eradication schemes increases the risk of developing their unwanted drug reactions, in particular, hepatotoxicity. As a means with potentially hepatoprotective activity, our attention was drawn to placental cryoextract.
 The purpose of the study. To characterize the gender aspects of the hepatotropic action of placenta cryoextract in tetrachloromethane hepatitis with background ethanol-induced cirrhosis and liver damage by antiulcer agents.
 Materials and methods. Research was conducted on 112 male and female rats. Tetrachloromethane hepatitis with background ethanol-induced liver cirrhosis was reproduced by injecting an oil solution of CCl4 in combination with a 5.0% ethanol solution for 45 days. Modulation of the content of sex hormones was achieved by surgical ovariectomy or testectomy.
 Results and discussion. Administration of esomeprazole, clarithromycin, and metronidazole in chronic liver damage in animals without changes in hormonal status led to a statistically significant (p=0.01) 27.6% increase in the level of bilirubin in homogenates of liver tissues in female rats (78.7±4.5 mmol/l) than in males. The greatest decrease in the content of bilirubin (41.7%, p<0.001) with the use of cryoextract of the placenta was noted in castrated female rats with simulated tetrachlormethone hepatitis with background ethanol-induced cirrhosis of the liver, which were injected with antiulcer agents.
 Conclusions. Administration of cryoextract of the placenta has a pronounced hepatoprotective effect in animals of both sexes. In male rats with no change in hormonal status with simulated liver injury and the administration of antiulcer drugs, placenta cryoextract therapy was more effective than in female rats, as indicated by a more than 1.6-fold decrease in the content of bilirubin in the liver homogenates of male rats (43.1%, p<0.001) than in females (27.4%, p<0.01).

Influence of NF-κB on the development of oxidative-nitrosative stress in the liver of rats under conditions of chronic alcohol intoxication

Alcohol-related liver disease is the most common cause of liver disease worldwide. The purpose of this work is the establishment of the influence of the transcription factor κB on the development of oxidative-nitrosative stress in the liver of rats under conditions of chronic alcohol intoxication. The experiments were performed on 24 male Wistar rats weighing 180-220 g. The animals were divided into 4 groups of 6 animals: control; animals, which were administered NF-κB inhibitor, namely ammonium pyrrolidinedithio­carbamate (PDTC) at a dose of 76 mg/kg 3 times a week; animals, on which we simulated alcoholic hepatitis and group of combination of alcoholic hepatitis and NF-κB inhibitor. We determined in rat liver homogenate the following biochemical parameters: the activi­ty of NO synthase isoforms, superoxide dismutase and catalase activity, the concentration of malonic dialdehyde, the concentration of peroxynitrite, nitrites and nitrosothiols, concentration of sulfide anion and superoxide anion radical production. Chronic alcohol intoxication led to increased production of reactive oxygen and nitrogen species on the background of decreased antioxidant activity, thus intensifying lipid peroxidation in the liver. Blockade of the transcription factor κB during chronic alcohol intoxication despite an increase in antioxidant activity and decrease of reactive oxygen and nitrogen species production did not ameliorate oxidative damage to the liver. Blockade of activation of nuclear transcription factor κB in rat liver by PDTC reduced the risk of oxidative damage to hepatocytes, but did not reduce the risk of developing nitrosative damage to hepatocytes. Keywords: chronic alcohol intoxication, NF-κB, oxidative-nitrosative stress, PDTC, rat liver

Rats with Long-Term Cholestasis Have a Decreased Cytosolic but Maintained Mitochondrial Hepatic CoA Pool.

Previous studies showed that rats with long-term bile duct ligation have reduced coenzyme A stores per g of liver but maintained mitochondrial CoA stores. Based on these observations, we determined the CoA pool in the liver homogenate, liver mitochondria, and liver cytosol of rats with bile duct ligation for 4 weeks (BDL rats, n = 9) and sham-operated control rats (CON rats, n = 5). In addition, we tested the cytosolic and mitochondrial CoA pools by assessing the metabolism of sulfamethoxazole and benzoate in vivo and of palmitate in vitro. The hepatic total CoA content was lower in BDL than CON rats (mean ± SEM; 128 ± 5 vs. 210 ± 9 nmol/g), affecting all subfractions equally (free CoA (CoASH), short- and long-chain acyl-CoA). In BDL rats, the hepatic mitochondrial CoA pool was maintained, and the cytosolic pool was reduced (23.0 ± 0.9 vs. 84.6 ± 3.7 nmol/g liver; CoA subfractions were affected equally). The urinary excretion of hippurate after i.p. benzoate administration (measuring mitochondrial benzoate activation) was reduced in BDL rats (23.0 ± 0.9 vs. 48.6 ± 3.7% of dose/24 h), whereas the urinary elimination of N-acetylsulfamethoxazole after i.p. sulfamethoxazole administration (measuring the cytosolic acetyl-CoA pool) was maintained (36.6 ± 3.0 vs. 35.1 ± 2.5% of dose/24 h BDL vs. CON rats). Palmitate activation was impaired in the liver homogenate of BDL rats but the cytosolic CoASH concentration was not limiting. In conclusion, BDL rats have reduced hepatocellular cytosolic CoA stores, but this reduction does not limit sulfamethoxazole N-acetylation or palmitate activation. The hepatocellular mitochondrial CoA pool is maintained in BDL rats. Impaired hippurate formation in BDL rats is explained best by mitochondrial dysfunction.

Effects of adenosine N1-Oxide and pioglitazone on inflammatory and antioxidant state in sepsis caused by experimental cecal puncture in rat.

Sepsis is an acute condition caused by the systemic inflammatory response syndrome to an infection. There are very few drugs that could improve the severe conditions in patients with sepsis. Hence, it is important to consider different treatment options. In this survey, we studied the effect of adenosine N1-oxide (ANO) and pioglitazone on rats with cecal ligation and perforation (CLP). They were randomly divided to four groups (n = 10) including Group A: as control group receiving normal saline, Group B: the rats with CLP as surgical control group, Group C: the rats receiving ANO, and Group D: the rats receiving pioglitazone. Interleukin (IL) -6, IL-1β, tumor necrosis factor alpha (TNF-α), nitric-oxide (NO) in serum blood and superoxide dismutase (SOD), catalase (CAT) malondialdehyde, (MDA) and myeloperoxidase (MPO) in liver and spleen homogenates were measured. The amount of antioxidant enzymes in the spleen and liver, and finally cell viability and rats' survival were investigated. The measurement of blood serum nitric-oxide and survival of all groups of rats were also performed. The results indicated that both drugs could cause a decrease in IL-1β, IL-6, TNF-α and NO in rat blood serum and MDA and MPO in the liver and spleen homogenates, however, a significant increase in SOD and CAT in the liver and spleen homogenates in rats that received ANO and pioglitazone was observed compared to rats with CLP group. Cell viability and rats' survival were significantly improved in rats that received ANO and pioglitazone compared to rats with CLP group. Adenosine N1-oxide showed stronger and more effective effects.

Riboflavin compounds show NAD(P)H dependent quinone oxidoreductase-like quinone reducing activity.

NAD(P)H-dependent quinone oxidoreductase (NQO) is an essential enzyme in living organisms and cells protecting them from oxidative stress. NQO reduces coenzyme Q (CoQ) using NAD(P)H as an electron donor. In the present study, we searched for coenzyme Q10 reducing activity from fractions of gel filtration-fractionated rat liver homogenate. In addition to the large-molecular-weight fraction containing NQO, CoQ10 reducing activity was also detected in a low-molecular-weight fraction. Furthermore, dicumarol, a conventional inhibitor of NQO1 (DT diaphorase), did not inhibit the reduction but quercetin did, suggesting that the activity was not due to NQO1. After further purification, the NADH-dependent CoQ10-reducing compound was identified as riboflavin. Riboflavin is an active substituent of other flavin compounds such as FAD and FMN. These flavin compounds also reduced not only CoQ homologues but also vitaminK homologues in the presence of NADH. The mechanism was speculated to work as follows: NADH reduces flavin compounds to the corresponding reduced forms, and subsequently, the reduced flavin compounds immediately reduce bio-quinones. Furthermore, the flavin-NADH system reduces CoQ10 bound with saposin B, which is believed to function as a CoQ transfer protein in vivo. This flavin-dependent CoQ10 reduction, therefore, may function in aqueous phases such as the cell cytosol and bodily fluids.

Pharmacological study of original extracts of corn silk

Aim: the aim of the research is a pharmacological study of the original extracts of corn silk. Materials and methods. The object of the study were 4 original extracts of corn silk, obtained by extraction with water and ethanol of different concentrations, namely: aqueous extract of corn silk (ACSE), extract of corn silk, extracted with ethanol 30 % (ECSE 30 %) , extract of corn silk, extracted with ethanol 50 % (ЕCSЕ 50 %), extract of corn silk, extracted with ethanol 70 % (ЕCSЕ 70 %). A screening study of the antioxidant properties of corn silk extracts was carried out in vitro using models of spontaneous and ascorbate-induced lipid peroxidation (LPO) in rat liver homogenate. Results. As can be seen from the given data, ECSE 50 % at a dose of 20 mg/kg showed a pronounced protective effect on the hepatotoxicity of TСM. Its use led to a probable restoration of the bile-forming function of the liver against the background of tetrachloromethane hepatitis: in response to an increase in the cholesterol content (by 43 %, p<0.05), the content of bile acids increased 2 times (p<0.05), as a result of which cholate -cholesterol ratio approached the level of indicators of intact animals, the rate of bile secretion normalized. Based on the analysis of the results of the study, it can be concluded, that in terms of bile-forming and choleretic activity on the tetrachloromethane hepatitis model, ECSE 50 % was not inferior to silibor and superior to quercetin (р<0.05). Conclusions. Biologically active substances of corn silk affect not only the diffusion and filtration processes of the liver parenchyma, but also the biosynthesis and transport of its organic components, that is, they affect the bile-forming function

Comparative antioxidant and hepatoprotective potential quercetin and corycavidine from <em>Hedyotis corymbosa </em>(L.) Lam. and <em>Solanum xanthocarpum</em> Schrad & Wendl

Liver disease is the major health issues in current era. Antioxidants play the essential role in hepatoprotection by protecting the hepatic cells against free radicals. Flavonoids and alkaloids are the essential plants bioactive that play the major role in the antioxidant system. In our project we used diamond flower Hedyotis corymbosa (L.) Lam. and Yellow berried Nightshade Solanum xanthocarpum Schrad & Wendl., and both plants have major phytoconstituent which acts as antioxidants. Thus, the aim of the current study validates the isolation, characterization, and determination of in vivo antioxidant and hepatoprotective consequences of corycavidine and quercetin that were isolated from Diamond flower Hedyotis corymbosa (L.) Lam. and Yellow berried Nightshade Solanum xanthocarpum Schrad & Wendl.. The study intent to isolate and identify the antioxidant and hepatoprotective agent from two different plants and compare their hepatoprotective potential to obtain the most effective liver protective phytoconstituent. Quercetin was isolated from S. xanthocarpum by column chromatography employing n-butanol: acetic acid: H2O (2:2:6) as a solvent system, however, corycavidine was isolated from H. corymbosa by column chromatography employing chloroform: methanol: diethylamine (4:1:2.2) as mobile phase. Structural illustrations were confirmed by UV, FT-IR, 1H-NMR, and 13C-NMR, and mass spectroscopy. Both the phytoconstituents, corycavidine and quercetin, were explored for their antioxidant potential by investigating CAT, SOD GSH, and LPO in liver homogenates of experimental rats. Additionally, the in vivo hepatoprotective effect was examined against simvastatin (20 mg/kg, p.o.), which induced hepatotoxicity in experimental rodents. The liver protective activity was computed by determining distinct biochemical parameters like SGOT, SGPT, ALP, bilirubin, total protein, cholesterol and urea along with hematological parameters and histopathological studies. The results of spectroscopic methods confirmed that the isolated phytochemical constituent from the H. corymbosa is corycavidine, a benzylisoquinoline alkaloid, however from S. xanthocarpum is quercetin a flavonoid. Both phytoconstituents significantly (P <0.05 P <0.001) and dose-dependently reversed simvastatin induced elevated levels of SGOT, SGPT, cholesterol, urea, total bilirubin and restored the total protein and albumin level in experimental rats. Furthermore, it also signifies the blood parameters at a dose of 50 and 100 mg/kg and restored the body protection system. The histological examination exhibited that corycavidine and quercetin at a dose of 100 mg/kg showed regeneration of hepatocytes around the central vein with nearly normal liver architecture. The results expressed the hepatoprotective outcome of quercetin is preeminent than corycavidine and therefore, scientifically validates its traditional application.

EFFECT OF CURCUMIN ON THE DEVELOPMENT OF OXIDATIVE-NITROSATIVE STRESS IN THE LIVER TISSUES OF RATS EXPOSED TO MODELED METABOLIC SYNDROME UNDER ROUND-THE-CLOCK LIGHTING

The aim of this study is to evaluate the effect of curcumin, a natural polyphenol, on indicators of oxidative-nitrosative stress in the liver of rats exposed to modeled metabolic syndrome (MS) under 24-hour lighting. The experiments were performed on 21 Wistar male white rats weighing 215-255 g. The animals were divided into 3 groups of 7 animals in each. The animals of the 1st group (control I) were kept on a standard vivarium diet and an ordered alternation of light / dark periods. The rats of the 2nd group were used for modeling diet-induced MS (a 2-month carbohydrate-lipid diet including 20% aqueous fructose solution for drinking and appropriate food) under the 24-hour lighting (control II). The rats of the 3rd group received curcumin (Sigma-Aldrich, Inc., USA) intragastrically through a gavage in a dose of 200 mg/kg daily during the 24-hour lighting exposure. We investigated the liver homogenate of rats to assess the rate of generation of the superoxide anion radical (•О), the activity of total NO synthase (NOS), its constitutive and inducible isoforms (cNOS, iNOS), and the content of peroxynitrites of alkali and alkaline earth metals. The administration of curcumin under the experimental conditions significantly suppresses the •О production in liver tissues by microsomal monooxygenases and NOS by 40.1%, by the mitochondrial respiratory chain by 34.4%, and by leukocyte NADPH oxidase by 32.1%; the activity of NOS and iNOS by 39.0% and 41.6%, the concentration of peroxynitrites of alkaline and alkaline earth metals by 23.8 compared to the values in the 2nd group. The activity of cNOS under these conditions was 2.79 times higher than the respective results in the II control, and the coupling index of this isoenzyme was 4.3 times higher. The main conclusion that can be drawn is that the administration of curcumin in the rat model of MS under the round-the-clock lighting significantly reduces the signs of oxidative-nitrosative stress in the liver tissues.

N-(9,10-Dioxo-9,10-dihydroanthracen-1(2)-yl)-2-(R-thio) Acetamides: Synthesis, Antioxidant and Antiplatelet Activity.

The synthesis of new N-(9,10-dioxo-9,10-dihydroanthracen-1(2)-yl)-2-(R-thio) acetamides was carried out using reaction of 2-chloro-N-(9,10-dioxo-9,10-dihydroanthracene-1(2)-yl)acetamides with functionalized thiols in the presence of potassium carbonate in N,N-dimethylformamide (DMF) at room temperature. Evaluation of the synthesized compounds on such indicators of radical scavenging activity as lipid peroxidation (LP) and oxidative modification of proteins (OMP) in vitro in rat liver homogenate was carried out. It was determined that the compounds with a substituent in the first position of anthracedione core showed better antioxidant properties than their isomers with a substituent in the second position. The compounds 6 and 7 with the best indicators of radical-scavenging activity were determined. Antioxidant effect in OMP processes was also determined for compound10. The antiplatelet activity study in vitro revealed compound 10 with the inhibited effect of ADP-induced aggregation.

PECULIARITIES OF PRENATAL INFLUENCE OF A NUTRITIONAL FACTOR ON THE STRUCTURAL AND FUNCTIONAL STATE OF THE LIVER OF NEWBORN RATS

Disease of the digestive system occupies one of the first places in the structure of morbidity and mortality in the population of Ukraine and EU countries. The alimentary factor (diet with an excess or deficiency of nutrients) is of leading importance Among the factors that cause liver damage in the mother-fetus system. The purpose of this study was to establish the effect of excess or insufficient content of nutrients in the mother's diet on the structural and functional state of the liver of newborn rats. Materials and Methods. To achieve this goal, 20 female rats of the WAG population were used; they were divided into 3 groups: group 1 (control) - rats were in standard vivarium conditions and received a basic diet; animals of the 2nd group received a diet with excess nutrients; rats of the 3rd group received a nutrient deficient diet. The offspring of rats were hatched from experiment through decapitation straightaway after birth. Was a complex of morphological and biochemical studies of liver tissue was carried out. Results. When analysing micro preparations of liver tissue, a similarity in the nature of the induced changes in the organ rats of the 2nd and 3rd groups in the form of moderately pronounced discomplexation of beamed-radiary structure and expansion sinusoids were revealed. The difference was that in rats of the 2nd group, mainly around the zone of portal tracts, small extramedullary hematopoiesis foci; and in rats of the 3rd group, multiple small extramedullary hematopoiesis foci that indicated more pronounced hypoxia were determined. Thus, it can be noted that the greatest damage to the fetus liver was caused by nutritional deficiency in the mother's diet. When studying the fractional composition of lipids in liver homogenates of newborn rats, the following dynamics of changes was discovered: in animals of the 2nd group - an increase of cholesterol and triglycerides levels, with a decrease of PL level, and in rats of the 3rd group, a decrease in almost all fractions of lipids cholesterol, triglycerides, and NEF A (nonesterified fatty acid). The obtained data suggested that, most likely, such dynamics of changes in lipid metabolism parameters is associated with the inclusion of epigenetic programming mechanisms in the mother-fetus complex. Conclusions. Therefore, based on conducted research, we can do conclusion about the negative impact of the alimentary factor (nutrient deficiency) on the structural and functional state of the liver of newborn offspring of rats.

Effects of Microencapsulated Ferulic Acid or Its Prodrug Methyl Ferulate on Neuroinflammation Induced by Muramyl Dipeptide.

Ferulic acid (Fer) is known for its antioxidant and anti-inflammatory activities, which are possibly useful against neurodegenerative diseases. Despite the ability of Fer to permeate the brain, its fast elimination from the body does not allow its therapeutic use to be optimized. The present study proposes the preparation and characterization of tristearin- or stearic acid-based solid lipid microparticles (SLMs) as sustained delivery and targeting systems for Fer. The microparticles were produced by conventional hot emulsion techniques. The synthesis of the methyl ester of Fer (Fer-Me) allowed its encapsulation in the SLMs to increase. Fer-Me was hydrolyzed to Fer in rat whole blood and liver homogenate, evidencing its prodrug behavior. Furthermore, Fer-Me displayed antioxidant and anti-inflammatory properties. The amount of encapsulated Fer-Me was 0.719 ± 0.005% or 1.507 ± 0.014% in tristearin or stearic acid SLMs, respectively. The tristearin SLMs were able to control the prodrug release, while the stearic acid SLMs induced a significant increase of its dissolution rate in water. Jointly, the present results suggest that the tristearin SLMs loaded with Fer-Me could be a potential formulation against peripheral neuropathic pain; conversely, the stearic acid SLMs could be useful for Fer-Me uptake in the brain after nasal administration of the formulation.

A Comparative Study of Quercetin-Loaded Nanocochleates and Liposomes: Formulation, Characterization, Assessment of Degradation and In Vitro Anticancer Potential.

Quercetin, a flavonoid, has antioxidant and anti-inflammatory properties and the potential to inhibit the proliferation of cancer, but its therapeutic efficacy is lowered due to poor solubility and bioavailability. Quercetin-loaded nanocochleates (QN) were developed using a trapping method by the addition of calcium ions into preformed negatively charged liposomes (QL) prepared by a thin-film hydration method. Liposomes were optimized by varying the concentration of Dimyristoyl phosphatidyl glycerol and quercetin by applying D-optimal factorial design using Design-Expert® software. Stable rods were observed using TEM with an average particle size, zeta potential and encapsulation efficiency of 502 nm, −18.52 mV and 88.62%, respectively, for QN which were developed from spherical QL showing 111.06 nm, −40.33 mV and 74.2%, respectively. In vitro release of quercetin from QN and QL was extended to 24 h. Poor bioavailability of quercetin is due to its degradation in the liver, so to mimic in vivo conditions, the degradation of quercetin released from QL and QN was studied in the presence of rat liver homogenate (S9G) and results revealed that QN, due to its unique structure, i.e., series of rolled up solid layers, shielded quercetin from the external environment and protected it. The safety and biocompatibility of QL and QN were provenby performing cytotoxicity studies on fibroblast L929 cell lines. QN showed superior anticancer activity compared to QL, as seen for human mouth cancerKB cell lines. Stability studies proved that nanocochleates were more stable than liposomal formulations. Thus, nanocochleates might serve as pharmaceutical nanocarriers for the improved efficacy of drugs with low aqueous solubility, poor bioavailability, poor targeting ability and stability.

Anticancer Cytotoxic Activity of Bispidine Derivatives Associated with the Increasing Catabolism of Polyamines.

Polyamine (PA) catabolism is often reduced in cancer cells. The activation of this metabolic pathway produces cytotoxic substances that might cause apoptosis in cancer cells. Chemical compounds able to restore the level of PA catabolism in tumors could become potential antineoplastic agents. The search for activators of PA catabolism among bicyclononan-9-ones is a promising strategy for drug development. The aim of the study was to evaluate the biological activity of new 3,7-diazabicyclo[3.3.1]nonan-9-one derivatives that have antiproliferative properties by accelerating PA catabolism. Eight bispidine derivatives were synthetized and demonstrated the ability to activate PA catabolism in regenerating rat liver homogenates. However, only three of them demonstrated a potent ability to decrease the viability of cancer cells in the MTT assay. Compounds 4c and 4e could induce apoptosis more effectively in cancer HepG2 cells rather than in normal WI-38 fibroblasts. The lead compound 4e could significantly enhance cancer cell death, but not the death of normal cells if PAs were added to the cell culture media. Thus, the bispidine derivative 4e 3-(3-methoxypropyl)-7-[3-(1H-piperazin-1-yl)ethyl]-3,7-diazabicyclo[3.3.1]nonane could become a potential anticancer drug substance whose mechanism relies on the induction of PA catabolism in cancer cells.

Global analysis of qualitative and quantitative metabolism of Notoginsenoside R1 in rat liver-brain-gut axis based on LC-IT-TOF/MS combing mMDF strategy

The metabolism study of active components for traditional Chinese medicine (TCM) in target organs is conducive to clarify the authentic active ingredients. Notoginsenoside R1 (NG-R1), one of the bioactive components of Panax notoginsenoside (PNS), is commonly acknowledged as the characteristic marker of PNS. However, the metabolism of NG-R1 in target organs has not been clarified yet due to the lack of robust technique and approach. The present study aimed to develop a reliable and efficient strategy and technology for revealing the qualitative and quantitative metabolism of active components of TCMs in target organs, and to clarify the biotransformation of NG-R1 in liver-brain-intestinal axis. The metabolic transformation of NG-R1 in the brain gut axis was investigated in the in vitro incubation system of fresh rat brain, liver homogenate, and intestinal flora. To quickly lock the target metabolites, we set the mass defect filter (MDF) in different ranges to screen metabolites with different molecular weight (MW). This strategy was defined as multi-stage MDF (mMDF). In addition, we performed relative quantitative analysis on all metabolites according to the peak area acquired by LC-IT-TOF/MS to overcome the challenge that metabolites are difficult to be quantified due to the lack of standards. When MDF was set at 0.50 to 0.65 to screen metabolites with MW of 900 to 1200 Da, 6 novel metabolites were quickly found, and then identified as glucuronic acid binding, oxidation, dehydrogenation, methylation and hydrogenation products according to their LC and MS characteristics. When setting MDF at 0.42 - 0.52, 6 metabolites with MW of 600 to 900 Da were effectively screened and identified as Rg1, NG-R2, Rh1, Rg1+CH2+2H and Rg1+CH2. To screen the metabolites with MW of 300 to 600 Da, MDF was set at 0.25 - 0.42, and 4 novel metabolites were screened rapidly. The results of quantitative metabolism suggested that intestinal flora was the main metabolic site of NG-R1 in rat, and more than 60% of NG-R1 was converted to Rg1 by deglycosylation in the intestinal flora. The mMDF strategy can significantly improve the research efficiency of qualitative metabolism of saponins. Although NG-R1 could be transformed into a variety of metabolites in rat liver and brain homogenate, it still existed mainly in prototype form. In the rat flora, NG-R1 mainly existed in the form of deglycosylated metabolite Rg1.

E161111 is an ultra-short-acting etomidate analogue with stable haemodynamics that elicits only slight adrenocortical suppression in rats.

PurposeWe report on a novel ultra-short-acting etomidate analogue, E161111, which has the same primary metabolite as etomidate.MethodsThe metabolic rate of E161111 was determined in rat plasma and liver homogenate. Rats were infused for 30 or 60 min to maintain light sedation at Richmond Agitation-Sedation Scale (RASS) for −2 to 0 score. Mean arterial pressure (MAP) was monitored during 30 min infusion. The serum corticosterone was determined during and 3 h after infusion as a measure of adrenocortical function.ResultsE161111 was not detected in rat plasma at 1 min (t1/2 = 6.69 ± 0.07 s) and in rat liver homogenates at 5 min (t1/2 = 10.20 ± 3.76 s); its main metabolic product was etomidate acid. The recovery time from loss of righting reflex (LORR) was 4.3 ± 1.5 min after 1-h infusion of E161111. During 30 min infusion, E161111 did not cause MAP changes. The stimulated serum corticosterone levels after 1-h infusion of E161111 were significantly higher than that after 1-h infusion of etomidate at all time points tested for the 3 h study.ConclusionsE161111 was metabolised rapidly, the metabolites were same as etomidate, and the recovery time after 1-h infusion was short. It elicited haemodynamic stability and milder suppression of corticosterone than that elicited by etomidate.

Synthesis and Antioxidant Activity of New Catechol Thioethers with the Methylene Linker.

Novel catechol thio-ethers with different heterocyclic substituents at sulfur atom were prepared by reacting 3,5-di-tert-butyl-6-methoxymethylcatechol with functionalized thiols under acidic conditions. A common feature of compounds is a methylene bridge between the catechol ring and thioether group. Two catechols with the thio-ether group, bound directly to the catechol ring, were also considered to assess the effect of the methylene linker on the antioxidant properties. The crystal structures of thio-ethers with benzo-thiazole moieties were established by single-crystal X-ray analysis. The radical scavenging and antioxidant activities were determined using 2,2′-diphenyl-1-picrylhydrazyl radical test, ABTS∙+, CUPRAC (TEAC) assays, the reaction with superoxide radical anion generated by xanthine oxidase (NBT assay), the oxidative damage of the DNA, and the process of lipid peroxidation of rat liver (Wistar) homogenates in vitro. Most catechol-thioethers exhibit the antioxidant effect, which varies from mild to moderate depending on the model system. The dual anti/prooxidant activity characterizes compounds with adamantyl or thio-phenol substituent at the sulfur atom. Catechol thio-ethers containing heterocyclic groups (thiazole, thiazoline, benzo-thiazole, benzo-xazole) can be considered effective antioxidants with cytoprotective properties. These compounds can protect molecules of DNA and lipids from the different radical species.