Liver GSH Content Research Articles

Reparative potentials of commensal-derived cellular components in juvenile Lateolabrax maculatus fed high soybean meal diet: Insight into growth performance, immunity, antioxidant capacity and liver health

An eight-week feeding trial was designed to estimate the effects of commensal Bacillus siamensis LF4-derived whole cell wall (CW), cell wall protein (CWP), lipoteichoic acid (LTA) and peptidoglycan (PGN) supplementation in a high soybean meal (SM) diet on the growth, immunity and liver health of spotted seabass (Lateolabrax maculatus). Fish continuously fed low SM (containing 16 % SM) and high SM (containing 40 % SM) diets were served as positive (LSM) and negative (HSM) control, respectively. After feeding high SM diet for 28 days, fish were fortified with B. siamensis LF4-derived CW, CWP, LTA or PGN until 56 days. The results showed that CW, LTA, and PGN application significantly increased FBW, WGR, and SGR (P<0.05), but not affect CF, VSI, HSI, IFR, ILI, and ISI (P＞0.05) in spotted seabass (L. maculatus) fed with high SM diet. Distal intestinal protease activity reduction caused by dietary high SM inclusion was significantly repaired when spotted seabass (L. maculatus) fortified with LTA and PGN (P<0.05). CW, LTA and PGN administration effectively increased serum immunity by enhancing ACP activity and C3, C4, IgM contents and reduced IL-1β content in serum of spotted seabass (L. maculatus) fed high SM diet (P<0.05). Significantly decreased serum MDA content in groups CW, LTA and PGN and significantly increased liver GSH content in group CWP were observed as compared to those in group HSM (P<0.05). Dietary CW, LTA and PGN supplementation, rather than CWP, repaired SM-induced liver injury by improving liver morphology and significantly reducing GOT and GPT activities (P<0.05). In summary, CW, LTA and PGN derived from B. siamensis LF4 effectively repaired SM-induced growth, feed utilization, immunity suppression and liver injury in spotted seabass (L. maculatus).

Pleurotus eryngii root waste and soybean meal co-fermented protein improved the growth, immunity, liver and intestinal health of largemouth bass (Micropterus salmoides)

The present study aimed to evaluate the effect of king oyster mushroom (Pleurotus eryngii) root waste and soybean meal co-fermented protein (CFP) on growth performance, feed utilization, immune status, hepatic and intestinal health of largemouth bass (Micropterus salmoides). Largemouth bass (12.33 ± 0.18 g) were divided into five groups, fed with diets containing 0 %, 5 %, 10 %, 15 % and 20 % CFP respectively for 7 weeks. The growth performance and dietary utilization were slightly improved by the supplementation of CFP. In addition, improved immunoglobulin M (IgM) content and lysozyme activity in treatments confirm the enhancement of immunity in fish by the addition of CFP, especially in fish fed 20 % CFP (P < 0.05). Furthermore, CFP significantly improved liver GSH (glutathione) content in groups D10 and D15 (P < 0.05), and slightly improved total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity while slightly reduced malondialdehyde (MDA) content. Simultaneously, the upregulation of lipolysis-related genes (PPARα, CPT1 and ACO) expression and downregulation of lipid synthesis-related genes (ACC and DGAT1) expression was recorded in the group D20 compared with the control (P < 0.05), which were consistent with the decreased liver lipid contents, suggests that lipid metabolism was improved by CFP. In terms of intestinal structural integrity, ameliorated intestinal morphology in treatments were consistent with the upregulated Occludin, Claudin-1 and ZO-1 genes expression. The intestinal pro-inflammatory cytokines (TNF-α and IL-8) expression were suppressed while the anti-inflammatory cytokines (IL-10 and TGF-β) were activated in treatments. The expression of antimicrobial peptides (Hepcidin-1, Piscidin-2 and Piscidin-3) and intestinal immune effectors (IgM and LYZ) were slightly up-regulated in treatments. Additionally, the relative abundance of intestinal beneficial bacteria (Firmicutes) increased while the relative abundance of potential pathogenic bacteria (Fusobacterium and Proteobacteria) decreased, which indicated that the intestinal microbial community was well-reorganized by CFP. In conclusion, dietary CFP improves growth, immunity, hepatic and intestinal health of largemouth bass, these data provided a theoretical basis for the application of this novel functional protein ingredient in fish.

262 Weaning-Induced Changes in Tissue Cysteine Utilization in Pigs

Abstract Weaning stress in pigs is associated with low and variable feed intake, poor nutrient utilization, and high feed costs. Cysteine is a sulfur amino acid with key roles in pig production, but how cysteine metabolism and requirements are affected by weaning stress should be better defined. The objective of this study was to determine weaning-induced changes in tissue cysteine metabolism. Pigs (initial BW = 7.23 ± 0.68 kg) were either weaned at 21 d age and euthanized at 23 d age (W; n = 9) or remained with the sow and euthanized at 23 d age (NW; n = 9). Weaned pigs were fasted to simulate acute nutritional stress but had free access to water. At euthanasia, blood, liver, jejunum, and ileum tissues were collected. Plasma and tissue amino acids and amino thiols were analyzed by HPLC. The abundance of proteins central to liver cysteine utilization were determined by immunoblot and enzyme activity assays. Glutathione (GSH) concentrations were less in jejunum (1,379 versus 1,720 ± 70 nmol/g; P &amp;lt; 0.01) and ileum (1,497 versus 1,740 ± 74 nmol/g; P &amp;lt; 0.05) in W than NW pigs. However, liver GSH concentration was not different between W and NW pigs (P &amp;gt; 0.10). While cysteine in jejunum and ileum were not affected by weaning (P &amp;gt; 0.10), γ-glutamylcysteine (γ-GC), the immediate precursor of GSH, declined in both jejunum (9.7 versus 14.3 ± 1.4 nmol/g; P &amp;lt; 0.01) and ileum (6.4 versus 11.2 ± 0.8 nmol/g; P &amp;lt; 0.001) in W compared with NW pigs. In liver, cysteine concentration was greater in W compared with NW pigs (417 versus 298 ± 33 nmol/g; P &amp;lt; 0.05). Although liver cysteine was greater in W pigs, liver activity of glutamate cysteine ligase (GCL; 2.49 versus 4.15 ± 0.25 nmol γ-GC/(mg protein•min); P &amp;lt; 0.001) and glutathione synthetase (GSS; 1.54 versus 1.92 nmol GSH/(mg protein•min); P &amp;lt; 0.001), enzymes needed for GSH production, were less in W than NW pigs. Immunoblot analysis of the catalytic subunit of GCL (GCLC; P &amp;lt; 0.05) and modifier subunit of GCL (GCLM; P = 0.057) were also lower in W compared with NW pigs, but GSS was not different between groups (P &amp;gt; 0.10). While plasma taurine was lower in W compared with NW pigs (64 versus 128 ± 7 µmol/L, P &amp;lt; 0.001), liver taurine was greater in W compared with NW pigs (4,726 versus 2,250 ± 471 nmol/g; P &amp;lt; 0.01). Despite an increase systemic and liver availability of cysteine, the rate-limiting amino acid for GSH synthesis, small intestine GSH concentrations decline. This suggests that systemic cysteine is not effectively utilized for gut GSH production in pigs after weaning.

Effects of Bacillus halophilus on growth, intestinal flora and metabolism of Larimichthys crocea

The incorporation of probiotics into the diet of large yellow croaker has been demonstrated by several studies to confer partial disease resistance. Bacillus halophilic isolated from the intestinal flora was used to study its effects on performance growth indicators, intestinal tissue structure, intestinal flora and the metabolism of Larimichthys crocea. A total of 180 fishes with an initial body weight of (164.00 ± 54.00) g were fed diets with three different concentrations of Bacillus halophilic: 0 cfu/mL (FC0, control group), 108 cfu/mL (FC8, treatment group), and 1012 cfu/mL (FC12, treatment group). The results showed that there were no significant differences in specific growth rate among all groups (P > 0.05). Compared to the FC0 group, the final body weight and Weight gain rate were significantly higher in FC8 and FC12 groups (P < 0.05). The Survival of the FC12 group significantly improved (P < 0.05). Compared to the FC0 group, crude protein content in muscle of the FC8 group significantly increased (P < 0.05), crude fat content significantly increased in the FC12 group (P < 0.05), crude protein content in whole fish experimental groups significantly increased (P < 0.05), and ash content significantly increased in the FC8 group (P < 0.05). In terms of antioxidant ability, the content of LZM in blood increased significantly in the FC8 group (P < 0.05), GSH content in liver of the FC12 group increased significantly (P < 0.05), while the content of MDA and AKP in blood and liver had no significant difference (P > 0.05). At the level of intestinal structure, there were no significant differences in villus height, crypt depth and goblet cell number between control group and treatment groups (P > 0.05). At the phylum level, Firmicutes was the dominant phylum, and the genus level, Lactobacillus and Bacteroides were the dominant bacteria in FC8 and FC12. A total of 1070 metabolites were identified, among which lipid metabolites accounted for 46.7%. Metabolites were involved in six main ways, mainly related to the metabolism of amino acids and lipids. The correlation analysis between microbes and metabolites showed that the intestinal flora of Larimichthys crocea could promote the synthesis of metabolites, among which Bacteroides and Megamonas could promote the synthesis of beneficial metabolites such as amino acids and vitamins. Through this study, we found that Bacillus halophilic can significantly improve growth, the antioxidant immunity ability and promote the expression of growth related metabolites, with the FC12 group being the better successful.

Protective Effect of Chrysin Against Chlorpyrifos-induced Hepatotoxicity in Rats: Biochemical and Histopathological Approaches in a Sub-chronic Study.

Chrysin (CH) is one of the important natural flavonoids with antioxidant and anti-inflammatory activity. The aim of this study was to assess the protective effects of CH on biochemical indexes and histopathological changes in the liver of male Wistar rats exposed to chlorpyrifos (CPF). We induced sub-chronic toxicity in rats using CPF (10 mg/kg/day, orally) and administrated CH at 12.5, 25, and 50 mg/kg/day for 45 days. In this study, CPF increased liver enzyme activities compared with the control group (p < 0.05), and co-treated CH with CPF reduced them compared with the non-treated CPF group (p < 0.05). A significant reduction in the liver GSH concentration along with a significant elevation in the concentrations of MDA and NO in the CPF group was observed compared with the control group (p < 0.001). However, CH at a dose of 50 mg could reverse them nearly to the control group (p < 0.001). In the CPF, CPF + CH1, and CPF + CH2 groups, a marked (p < 0.05) increase was found in the serum concentration of IL-6 compared with the control animals. No significant changes were found in the IL-6 concentration of the CPF + CH3 compared with the controls. Moreover, the coadministration of CH plus CPF induced histopathological alterations in liver. These results suggest that CH attenuates hepatic enzymes and histopathological alterations induced by CPF via modulating oxidative stress and inflammatory indices in rats.

γ‐glutamylcysteine alleviates insulin resistance and hepatic steatosis by regulating adenylate cyclase and IGF‐1R/IRS1/PI3K/Akt signaling pathways

Type 2 diabetes mellitus (T2DM), a complex metabolism disease, which was characterized by metabolic disorders including hyperglycemia, has become a major health problem due to the increasing prevalence worldwide. γ-glutamylcysteine (γ-GC) as an immediate precursor of glutathione (GSH) was originally used for the treatment of sepsis, inflammation bowel disease, and senescence. Here, we evaluated the capacity of γ-GC on diabetes-related metabolic parameters in db/db mice and insulin resistance (IR) amelioration in cells induced by palmitic acid (PA). Our data suggested that γ-GC treatment decreased body weight, reduced adipose tissue size, ameliorated ectopic fat deposition in liver, increased the GSH content in liver, improved glucose control and other diabetes-related metabolic parameters in vivo. Moreover, in vitro experiments showed that γ-GC could maintain the balance of free fatty acids (FFAs) and glucose uptake through regulating the translocation of CD36 and GLUT4 from cytoplasm to plasma membrane. Furthermore, our finding also provided evidence that γ-GC could activate Akt not only via adenylate cyclase (AC)/cAMP/PI3K signaling pathway, but also via IGF-1R/IRS1/PI3K signaling pathway to improve IR and hepatic steatosis. Blocking either of two signaling pathways could not activate Akt activation induced by γ-GC. This unique characteristic ensures the important role of γ-GC in glucose metabolism. Collectively, these results suggested that γ-GC could serve as a candidate dipeptide for the treatment of T2DM and related chronic diabetic complications via activating AC and IGF-1R/IRS1/PI3K/Akt signaling pathways to regulate CD36 and GLUT4 trafficking.

Electroacupuncture promotes expression of glutathione related regulatory enzymes in ovary tissue of rats with diminished ovarian reserve

To observe the effect of electroacupuncture (EA) on ovarian function and expression of glutathione (GSH) related regulatory enzymes γ-glutamylcysteine synthetase (γ-GCS), glutathione reductase (GR) protein and gene in rats with diminished ovarian reserve (DOR), so as to explore its mechanisms underlying up-regulation of antioxidant stress ability. A total of 30 female SD rats with normal estrous cycle were randomly divided into blank control, model and EA groups, with 10 rats in each group. The DOR model was established by gavage of tripterygium wilfordii polyglycoside suspension (50 mg·kg-1·d-1) for 14 consecutive days, while the rats in the blank group were given equal volume of 0.9% sodium chloride solution. One hour after daily gavage, EA (1.0 mA, 100 Hz) was applied alternately to bilateral "Shenshu"(BL23), and "Zhongwan"(CV12)+"Guanyuan"(CV4) for 10 min, for 14 consecutive days. Estrous cycles of rats in each group were observed and recorded daily during intervention.After the intervention, H.E.staining was used to observe histopathological changes of the ovarian tissue. The contents of serum sex hormones ［follicle stimulating hormone (FSH), anti-mullerian hormone (AMH), estradiol (E2)］ and oxidative damage markers ［8-hydroxydeoxyguanosine (8-OHDG) and nitrotyrosine (NTY)］ were determined by ELISA. The contents of GSH and oxidized glutathione (GSSG) in the liver tissue were determined by colorimetry, and their ratios were calculated. Immunohistochemistry and real-time fluorescence quantitative PCR were used to detect the immunoactivity and gene expression levels of γ-GCS and GR in the ovarian tissues, respectively. Compared with the blank group, the model group had a marked increase in the disorder rate of estrous cycle, serum FSH, 8-OHDG and NTY contents (P<0.01) and a considerable decrease in the levels of serum AMH and E2, liver GSH and GSSG contents and GSH/GSSG ratio, ovarian optical density and cell number as well as the expression of γ-GCS and GR mRNAs (P<0.05, P<0.01). After EA intervention, the increase of the disorder rate of estrous cycle, serum FSH, 8-OHDG and NTY contents and the decrease of serum AMH and E2, liver GSH and GSSG contents and GSH/GSSG ratio, ovarian optical density and cell number of γ-GCS and GR as well as the expression of γ-GCS genes were all reversed (P<0.01, P<0.05). H.E. staining showed degenerative changes of the ovarian tissue, fewer follicles at every level and increase of atretic follicles, disarrangement and layer number decrease of granulosa cells, and atrophy of corpus luteum in the model group, which were relatively milder in the EA group. EA can improve ovarian function, and reduce oxidative stress damage in DOR rats, which may be associated with its functions in up-regulating the expression of γ-GCS and GR protein and gene in the ovarian tissue.

The effects of simvastatin and fenofibrate on malondialdehyde and reduced glutathione concentrations in the plasma, liver, and brain of normolipidaemic and hyperlipidaemic rats.

The objective of study was to investigate the effects of different doses of simvastatin and fenofibrate on malondialdehyde (MDA) and reduced glutathione (GSH) in the plasma, liver, and brain tissue of male normolipidaemic and hyperlipidaemic rats. Normolipidaemic (Wistar) rats were receiving 10 or 50 mg/kg a day of simvastatin or 30 or 50 mg/kg a day of fenofibrate. Hyperlipidaemic (Zucker) rats were receiving 50 mg/kg/day of simvastatin or 30 mg/kg/day of fenofibrate. Control normolipidaemic and hyperlipidaemic rats were receiving saline. Simvastatin, fenofibrate, and saline were administered by gavage for three weeks. In normolipidaemic rats simvastatin and fenofibrate showed similar and dose-independent effects on plasma and brain MDA and GSH concentrations. Generally, plasma and brain MDA decreased, while brain GSH concentration increased. In hyperlipidaemic rats simvastatin did not affect plasma and brain MDA and GSH concentrations but significantly decreased liver GSH. Fenofibrate decreased plasma and liver MDA but increased brain MDA. In both rat strains fenofibrate significantly decreased liver GSH concentrations, most likely because fenofibrate metabolites bind to GSH. Our findings suggest that simvastatin acts as an antioxidant only in normolipidaemic rats, whereas fenofibrate acts as an antioxidant in both rat strains.

Astaxanthin mitigates oxidative stress caused by microplastics at the expense of reduced skin pigmentation in discus fish

Microplastics (MPs) exposure generally triggers oxidative stress in fish species and vertebrate pigmentation is commonly influenced by oxidative stress, but MPs-induced oxidative stress on fish pigmentation and body color phenotype has not been reported. The aim of this study is to determine whether astaxanthin could mitigate the oxidative stress caused by MPs but at the expense of reduced skin pigmentation in fish. Here, we induced oxidative stress in discus fish (red skin color) by 40 or 400 items/L MPs under both astaxanthin (ASX) deprivation and supplementation. We found that lightness (L*) and redness (a*) values of fish skin were significantly inhibited by MPs under ASX deprivation. Moreover, MPs exposure significantly reduced ASX deposition in fish skin. The total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity in fish liver and skin were both significantly increased with the increase of MPs concentration, but content of glutathione (GSH) in fish skin showed a significant decrease. For ASX supplementation, the L*, a* values and ASX deposition were significantly improved by ASX, including the skin of MPs-exposed fish. The T-AOC and SOD levels changed non-significantly in fish liver and skin under the interaction of MPs and ASX, but ASX significantly reduced GSH content in fish liver. Biomarker response index indicated that ASX could improve the moderately altered antioxidant defense status of MPs-exposed fish. This study suggests that the oxidative stress caused by MPs was mitigated by ASX but at expense of reduced fish skin pigmentation.

Alteration of Glutathione and Antioxidant Status with Exercise in Unfed and Refed Rats

The influences of food deprivation and refeeding on glutathione (GSH) status, antioxidant enzyme activity and lipid peroxidation in response to an acute bout of exercise were investigated in the liver and skeletal muscles of male Sprague-Dawley rats randomly divided into three groups: starved for 48 h without refeeding; starved for 48 h and refed for 24 or 48 h. Half of each group of rats was exercised on a treadmill until exhaustion and killed immediately, whereas the other half group was killed at rest. Food-deprived rats had significantly lower liver GSH concentration and GSH:glutathione disulfide (GSSG) ratio. Malondialdehyde concentrations in the liver and skeletal muscle were both higher in the starved than in the refed rats (P < 0.05). Refed rats had significantly greater liver GSH level, γ-glutamylcysteine synthetase and glucose 6-phosphate dehydrogenase activities and plasma insulin concentration than unfed rats. Exercised 24- and 48-h refed rats had 27% and 31% lower liver GSH (P < 0.05), respectively, and a 21% lower GSH:GSSG ratio (P < 0.05) than their rested counterparts. Plasma insulin concentrations were significantly lower, whereas glucagon concentrations were greater in the exercised than in the rested rats. Muscle GSH concentration was significantly lower in the food-deprived than in the refed rats (P < 0.05) but was unaffected by exercise. Exercised 24-h refed rats had significantly elevated muscle GSSG concentration compared with rested rats, along with a higher GSH peroxidase and a lower γ-glutamyltranspeptidase activity (P < 0.05). These data indicate that both food deprivation-refeeding and exhaustive exercise influence liver and skeletal muscle glutathione status and that these changes may be controlled by hepatic glutathione synthesis and release due to hormonal stimulation.

Technical note: simultaneous determination of amino thiols in pig tissue by ultra-high performance liquid chromatography with fluorescence detection.

Sulfur amino acid nutrition and metabolism are linked to animal disease. While validated methods for the determination of amino thiol levels in plasma or serum are available, there is a dearth of validated methods for their measurement in tissue. A robust and reproducible ultra-high performance liquid chromatography method has been validated for the simultaneous determination of concentrations of cysteine (Cys), cysteinylglycine (CysGly), homocysteine (Hcys), γ-glutamylcysteine (γ-GluCys), and glutathione (GSH) in pig tissue. Tissue was homogenized and deproteinized with trichloroacetic acid. Amino thiols in the acid-soluble fraction of the tissue homogenate were reduced with tris-(2-carboxyethyl)-phosphine hydrochloride and derivatized with 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F). Amino thiols were resolved under reversed-phase gradient conditions on a Waters Acquity BEH C18 column (1.7 µm, 2.1 mm × 100 mm) within 4.5 min and detected with fluorescence. The peak area ratio of analyte to 2-mercaptopropionylglycine internal standard, added to external calibration standards and samples, was used to develop linear calibration curves. Linear calibrations were performed over the range of 15-1,500 nmol/g for Cys, CysGly, Hcys, and γ-GluCys and 150-15,000 nmol/g for GSH. Linearity, lower limit of detection, lower limit of quantitation, accuracy, precision, sample stability, and carryover were evaluated. We demonstrate excellent linearity for all analytes within their respective concentration range (r2 > 0.99) and excellent recovery of amino thiols from spiked samples (mean ± SD across tissues; Cys, 100.0 ± 2.2%; CysGly, 95.4 ± 5.1%; Hcys, 96.6 ± 2.0%; γ-GluCys, 102.2 ± 2.7%; and GSH, 100.6 ± 3.3%). The intra-day and inter-day precisions did not exceed 5% and 10%, respectively. Repeated freezing and thawing of tissue homogenate did not affect measured amino thiol concentrations, ABD-labeled amino thiols were stable for 1 wk after derivatization, and there was no sample carryover across consecutive injections. We confirm the identity of each ABD-labeled amino thiol with Orbitrap mass spectrometry. Finally, we apply the method to the determination of amino thiol concentrations in liver and jejunum tissues in newly weaned pigs and show that despite elevated Cys and maintained GSH concentrations in liver, both γ-GluCys and GSH decline in jejunum of weaned pigs.

Investigation of the effect of N-acetylcysteine on aluminum phosphide toxicity in rats

Aluminum Phosphide (ALP) is one of the most dangerous pesticides. When it comes into contact with water, it emits Phosphine (PH3) gas, which causes poisoning and death in many people. The purpose of this study is to look into the role of N-acetylcysteine in the treatment of aluminum phosphide toxicity in rats. In this study, 30 male Wistar rats were fed with aluminum phosphide orally. After 15 minutes, N-acetylcysteine was administered intraperitoneally. The antioxidant enzymes glutathione S-Transferase (GST), Superoxide Dismutase (SOD), Catalase (CAT), glutathione (GSH), Aspartate Aminotransferase (AST), Alanine Transaminase (ALT), and Alkaline Phosphatase (ALK) were studied in blood plasma. CAT, GST, and GSH concentrations in plasma, liver, and kidneys of rats infected with aluminum phosphide decreased, while AST, ALT, and ALK concentrations increased. The levels of all enzymes studied approached normal after N-acetylcysteine administration, and the rats survived for up to 12-15 hours. According to the findings of this study, N-acetylcysteine (NAC) at a dose of 10 mg/kg improves hepatic manifestations and prevents liver necrosis, so it can be considered a potential therapeutic agent in the treatment of this poisoning.

Dietary bile acids supplementation modulates immune response, antioxidant capacity, glucose, and lipid metabolism in normal and intrauterine growth retardation piglets.

Intrauterine growth retardation (IUGR) results in intestinal dysfunction contributing to metabolic syndrome and growth lag of piglets. Bile acid (BA) presents various bioactivities, including regulation roles in antioxidant, anti-inflammation, and glucose and lipid metabolism. Forty-eight weaned piglets were allocated to four groups in a 2 × 2 factorial arrangement with the effects of BA supplementation and IUGR challenge. Twenty-four IUGR piglets and 24 normal birth weight (NBW) piglets were allocated into two groups, respectively, including the control group fed with a basal diet, and the treatment group fed a basal diet supplemented with 400 mg/kg BA. The experiment lasted 28 days. The results indicated that BA improved liver and spleen indexes in IUGR piglets, whereas decreased blood RDW-CV and RDW-SD regardless of IUGR (P < 0.05). Dietary BA supplementation decreased plasma CAT activity and liver GSH concentration regardless of IUGR, whereas increased plasma GSH and liver H2O2 and decreased liver T-AOC in weaned piglets (P < 0.05). In addition, IUGR downregulated liver Nrf1 and Nrf2 expression levels, while BA supplementation upregulated the Nrf2 expression of liver in weaned piglets (P < 0.05). Dietary BA decreased (P < 0.05) jejunal GSH concentration and ileal CAT activity regardless of IUGR. Furthermore, IUGR upregulated (P < 0.05) jejunal SOD and CAT expression levels; however, dietary BA upregulated ileal Nrf1 (P < 0.05) and Keap1 (P = 0.07) expression levels in piglets regardless of IUGR. Moreover, IUGR upregulated the liver lipid synthesis (FAS) and downregulated HSL and SCD1 expression levels, while dietary BA downregulated liver FAS and SCD1 expression levels (P < 0.05). However, BA supplementation could enhance liver gluconeogenesis by upregulating (P < 0.05) the liver G6PC and PCK1 expression levels in the NBW piglets but not in the IUGR piglets. Collectively, these findings suggest that BA could regulate the redox status of weaned piglets by regulating the Nrf2/Keap1 pathway and improving liver glucose and lipid metabolism of IUGR piglets. These findings will provide a reference for the application of BA in swine production; moreover, considering the physiological similarity between pigs and humans, these findings will provide a reference for IUGR research in humans.

Biochemical determination of reduced glutathione concentration in pig liver and kidney in the presence of chlorinated aquatic humic materials

Disinfection of surface waters for human consumption and use often results in reaction between the disinfectant (generally chlorine) and organic substrates present at the source. These substrates are majorly in the form of aquatic humic materials. This work was designed to assess reduced glutathione (GSH) concentration in pig liver and kidney in the presence of chlorinated aquatic humic materials (CAHMs) using biochemical parameters. GSH concentration in pig liver and kidney was investigated following standard procedures using Ellman’s reagent. Effect of chlorinated local and foreign aquatic humic products on the GSH concentration of the two tissues was studied in line with standard methodologies. GSH concentration was found to be higher in pig liver (0.874 ± 0.003µmol/g) than the kidney (0.545 ± 0.002µmol/g). All the chlorinated aquatic humic samples studied were found to deplete the GSH concentration of the two tissues, with the more toxic and concentrated purified chlorinated foreign aquatic humics having higher depletion. Certain amount of GSH was also found to be left in both tissues after conjugation with the chlorinated aquatic humic samples which perhaps could be used for subsequent conjugation and detoxification of water pollutants by the tissues. In conclusion, the liver contains more GSH than the kidney. The levels of this antioxidant in pig liver and kidney were both reduced by the aquatic humic materials used. This study may therefore reemphasize earlier suggestions that the amino acids needed for cellular synthesis of GSH could be supplemented regularly through dietary sources.

Holly polyphenols attenuate liver injury, suppression inflammation and oxidative stress in lipopolysaccharide-challenged weaned pigs

ABSTRACT It has been proved that polyphenols have positive effects against inflammatory response and oxidative stress. We investigated whether dietary supplementation of the holly polyphenols (HP) might relieve a liver injury induced by lipopolysaccharide (LPS) in weaned pigs. Twenty-four weaned pigs were fed diets containing 0 or HP and saline or LPS challenge in 2×2 factorial arrangement of treatments. On d 17, pigs were intraperitoneally injected with an LPS or saline. Dietary HP supplementation reduced plasma AST and GGT activities and liver MDA levels, increased plasma ALB and TP and liver GSH concentrations, and ameliorated LPS-induced liver histopathological damage. HP supplementation reduced plasma and /or liver TNF-α, IL-6 and IL-1β levels, and liver mRNA expression of IL-6, TNF-α and COX2. Moreover, HP decreased the mRNA abundance of liver MyD88, TRAF6 and NOD1. These results suggest that HP supplementation attenuates liver injury induced by LPS in weaned pigs, and inhibits inflammatory response and oxidative stress.

The Influence of Dietary Zinc Oxide Nanoparticles on Zootechnical Performance and Biometric Indices of Growing New Zealand White Rabbits

This study was designed to compare the effects of dietary conventional inorganic Zinc oxide (ZnO) and nano-zinc oxide (ZnO-NPs) on the zootechnical performance, selected blood serum indices, antioxidant status, general health and production of rabbit meat enriched with zinc enriched with nano-zinc in growing New Zealand White rabbits (NZW). A total of 481 newly-born rabbits produced from two natural successive mating between 36 primiparous (NZW) Rabbit does and 12 bucks by the intensive breeding system were utilized throughout this study. Rabbits does and bucks were randomly distributed into four equal groups, the control positive group G1 was fed the basal diet supplemented with premix contain 50 ppm inorganic zinc in the form of ZnO; while the control negative G2 was fed on a basal diet supplemented with Zinc Free premix, meanwhile, G3 was fed on the basal diet supplemented with premix contain 25 ppm nano zinc as zinc oxide nanoparticles, however, G4 was fed on the basal diet supplemented with 50 ppm nano zinc as zinc oxide nanoparticles in the premix, Litters after weaning continued on the same diet like their dams till the age of selling at 62 days old. The results showed that the growing rabbits in (G4) supplemented with 50 mg ZnO-NPs/kg diet surpassing significance (P≤ 0.05) in all groups and achieved the best result regarding (birth weight(g), weaning weight(g), sealing weight (g) moreover lower Pre-weaning mortality %, and Post-weaning mortality%). Also, G4 showed significant improvement in FCR in comparison with other groups. Furthermore, showed highest significant increase in total protein and globulin, HDL, SOD, GSH, Catalase activity, and zinc concentration in serum, liver, and thigh tissue and also showed the highest significant decrease in AG ratio, creatinine, urea, uric acid, cholesterol, Triglyceride, LDL, VLDL, risk factor, MDA and NO level in G4 at 62 day age old, compared to other groups, while there was no significant difference in albumin level between all the experimental groups. In conclusion, supplementation of 50 mg ZnO-Npskg diet in the premix to the diet of growing NZW rabbit maximize the productive performance and improve FCR without any adverse effect.

Concentration of current-use pesticides in frogs from the Pampa region and correlation of a mixture toxicity index with biological effects

Contamination with current-use pesticides is frequently mentioned as a key factor in global amphibian declines although a limited number of studies have examined the mixture of pesticides accumulated by free-living frogs. This study examined the presence of 46 different pesticide residues in the muscle and kidney tissues of two frog species living in close association with row crops in the Pampa region of Argentina: The terrestrial Leptodactylus latinasus and the semi-aquatic Leptodactylus latrans. A total of 20 different pesticides were identified in frog tissues; chlorpyrifos-methyl, pirimiphos-methyl and acetochlor being the most frequently detected molecules. Overall, one or more pesticide residues (up to 12 in a single frog) were detected in 40–57 % of L. latrans. L. latinasus was found to present more pesticide detections than L. latrans. Interestingly, frog sampled in a natural reserve where no pesticides are applied presented an equivalent frequency of detections as frogs living near a crop. In L. latrans, the calculation of a pesticide toxicity index (PTI) permitted to highlight the existence of a strong positive correlation between PTI and liver GSH contents of females whereas, in males, PTI was negatively correlated with the perimeter of testicular seminiferous tubules. Males also presented near significant negative correlations between PTI and both body condition and the scaled fat index. These results indicate that frogs inhabiting agricultural regions are exposed to a complex and diffuse contamination by pesticide mixtures which is likely responsible for a number of biological effects that may be relevant at the population level.

β-Sitosterol Alters the Inflammatory Response in CLP Rat Model of Sepsis by Modulation of NFκB Signaling.

Purpose Sepsis originates from the host inflammatory response, especially to bacterial infections, and is considered one of the main causes of death in intensive care units. Various agents have been developed to inhibit mediators of the inflammatory response; one prospective agent is β-sitosterol (βS), a phytosterol with a structure similar to cholesterol. This study is aimed at evaluating the effects of βS on the biomarkers of inflammation and liver function in cecal ligation and puncture- (CLP-) induced septic rats. Methods Thirty male Wistar rats were divided equally into six groups as follows: sham, CLP, CLP+dexamethasone (DX, 0.2 mg/kg), CLP+βS (1 mg/kg), CLP+imipenem (IMI, 20 mg/kg), and CLP+IMI (20 mg/kg)+βS (1 mg/kg). Serum levels of IL-1β, IL-6, IL-10, AST, ALT, and liver glutathione (GSH) were assessed by ELISA. Liver expression levels of TNF-α and NF-κBi mRNAs were evaluated by RT-qPCR. Results Serum concentrations of IL-1β, IL-6, IL-10, ALT, and AST and mRNA levels of TNF-α and NF-κBi were all significantly higher in septic rats than in normal rats (p < 0.05). Liver GSH content was markedly lower in the CLP group than that in the sham group. βS-treated rats had remarkably lower levels of IL-1β, IL-6, IL-10, TNF-α, NF-κBi, AST, and ALT (51.79%, 62.63%, 41.46%, 54.35%, 94.37%, 95.30%, 34.87%, and 46.53% lower, respectively) and greater liver GSH content (35.71% greater) compared to the CLP group (p < 0.05). Conclusion βS may play a protective role in the septic process by mitigating inflammation. This effect is at least partly mediated by inhibition of the NF-κB signaling pathway. Thus, βS can be considered as a supplementary treatment in septic patients.

Pregabalin, a GABA Analogue Protects against Carbon Tetrachloride-Induced Oxidative Stress and Liver Injury in Rats

Pregabalin is a synthetic analogue of the neurotransmitter g-aminobutyric acid (GABA) and is used in the treatment of epilepsy and neuropathic pain. The aim of this study was to investigate the effect of pregabalin on toxic liver injury caused by the acute administration of carbon tetrachloride (CCl4). Rats were orally treated with CCl4 for two successive days either alone or along with intraperitoneal pregabalin at doses of 7 and 14 mg/kg. The control group received the vehicle (olive oil). Liver oxidative stress and damage were assessed by determining serum and/or liver tissue levels of malondialdehyde (MDA), nitric oxide (NO), reduced glutathione (GSH), DNA fragmentation, serum aminotransferases, paraoxonase-1 (PON-1), and hepatic histopathology. Results showed that CCl4 significantly (i) increased MDA and NO and decreased PON-1 in both serum and liver tissue, and (ii) decreased liver GSH content and induced marked hepatic DNA fragmentation. CCl4 treatment caused liver tissue injury as evidenced by significantly increased serum aminotransferases. In line with the above biochemical changes, the liver of CCl4-treated rats exhibited massive steatosis, vacuolar degeneration, cloudy swelling, and necrosis. In CCl4-treated rats, pregabalin given at the dose of 14 mg/kg significantly reduced the increments in MDA, NO, serum aminotransferases, and hepatic DNA fragmentation. Liver GSH was unaltered but hepatic and serum PON-1 activity increased after administering pregabalin which also improved, though not normalized, liver tissue histopathology. Collectively, these results suggest that the administration of pregabalin is associated with a reduction in experimental liver injury caused by CCl4. (First online: Apr 12, 2021)

Naringin and Hesperidin Counteract Diclofenac-Induced Hepatotoxicity in Male Wistar Rats via Their Antioxidant, Anti-Inflammatory, and Antiapoptotic Activities.

This study is aimed at evaluating the preventive effect and at suggesting the mode of actions of naringin and hesperidin and their combination in diclofenac-induced hepatotoxicity. Male Wistar rats, intraperitoneally injected with diclofenac sodium (3 mg/kg b.wt/day), were orally treated with naringin (20 mg/kg b.wt/day) and hesperidin (20 mg/kg b.wt/day) and their combination for 4 weeks. The administrations of naringin and hesperidin to diclofenac-injected rats led to a significant decrease in the elevated serum ALT, AST, LDH, ALP, GGT, total bilirubin, TNF-α, and IL-17 levels as well as liver lipid peroxidation and liver p53 and caspase-3 mRNA expressions. In contrast, serum IL-4 level, liver GSH content, and liver GPx and SOD activities increased. In association, diclofenac-induced deleterious histological alterations including hydropic degeneration, cytoplasmic vacuolization, apoptosis, and focal hepatic necrosis of hepatocytes associated with inflammatory cells' infiltration were remarkably improved by treatments with naringin and hesperidin. In conclusion, naringin, hesperidin, and their combination, which was the most potent, counteract diclofenac-induced liver injury via antioxidant, anti-inflammatory, and antiapoptotic actions. Thus, this study recommends the use of naringin and hesperidin or their combination to resolve the side effects of drugs like diclofenac on the liver.