Liver mRNA Expression Research Articles

Oral administration of trehangelin-A alleviates metabolic disorders caused by a high-fat diet through improvement of lipid metabolism and restored beneficial microbiota

The present study investigated whether or not the oral administration of trehangelin-A (THG-A) is effective for metabolic disorders caused by a high-fat diet, as we previously showed that the intraperitoneal administration of THG-A improved metabolic disorders caused by a high-fat diet. Mice received a control diet or high-fat diet for eight weeks. Concurrently, mice were orally administered 0.2 ml/mouse phosphate-buffered saline (PBS) or 1 or 10 mg/0.2 ml/mouse of THG-A once daily during the experiment. The weight gain caused by a high-fat diet was significantly suppressed by oral THG-A compared to a high-fat diet without THG-A. In addition, at eight weeks after starting the diet, the increased plasma total-cholesterol (T-CHO) and low-density lipoprotein-cholesterol (LDL-C) levels caused by a high-fat diet were significantly reduced by 10 mg/mouse THG-A and tended to attenuated by 1 mg/mouse THG-A. The LDL receptor and CYP7A1 mRNA expression in liver associated with lipid metabolism for reducing plasma LDL-C levels was significantly enhanced by oral THG-A. In contrast, oral THG-A exerted no marked effects on mice fed the control diet. The dysbiosis of a high-fat diet fed mice, which is in the form of an increased Firmicutes-to-Bacteroidetes ratio, also recovered, and the high-fat diet induced decreased levels of Bacteroides and Akkermansia genera, which are beneficial microbiota against metabolic disorders, were also restored by oral THG-A. These results indicate that oral THG-A administration acts on metabolic disorders by improving the lipid metabolism and restoring beneficial microbiota to resolve high-fat diet induced dysbiosis.

Effects of exogenous lactate administration on fat metabolism and glycogen synthesis factors in rats.

[Purpose]Lactate has several beneficial roles as an energy resource and in metabolism. However, studies on the effects of oral administration of lactate on fat metabolism and glycogen synthesis are limited. Therefore, the purpose of the present study was to investigate how oral administration of lactate affects fat metabolism and glycogen synthesis factors at specific times (0, 30, 60, 120 min) after intake.[Methods]Male Sprague Dawley (SD) rats (n = 24) were divided into four groups as follows: the control group (0 min) was sacrificed immediately after oral lactate administration; the test groups were administered lactate (2 g/kg) and sacrificed after 30, 60, and 120 min. Skeletal muscle and liver mRNA expression of GLUT4, FAT/CD36, PDH, CS, PC and GYS2 was assessed using reverse transcription-polymerase chain reaction.[Results]GLUT4 and FAT/CD36 expression was significantly increased in skeletal muscle 120 min after lactate administration. PDH expression in skeletal muscle was altered at 30 and 120 min after lactate consumption, but was not significantly different compared to the control. CS, PC and GYS2 expression in liver was increased 60 min after lactate administration.[Conclusion]Our results indicate that exogenous lactate administration increases GLUT4 and FAT/CD36 expression in the muscle as well as glycogen synthase factors (PC, GYS2) in the liver after 60 min. Therefore, lactate supplementation may increase fat utilization as well as induce positive effects on glycogen synthesis in athletes.

1720-P: The Modulation in Glucose Metabolism by B1 Kinin Receptor Does Not Depend in Hepatic Insulin Signaling

Background: The kinin B1 receptor (B1R), an inducible effector of Kallikrein-Kinin System during inflammation, has been blamed also to influence glucose metabolism and leptin sensitivity. Aim: To investigate the effects of cafeteria diet (CAF) on hepatic glucose metabolism in B1R knockout mice (B1RKO). Methods: Two months old male C57Bl/6 wild type and B1RKO mice were fed with standard chow (SC) or CAF ad libitumfor 14 weeks (WT-SC n=7; B1RKO-SC n=8; WT-CAF n=7; B1RKO-CAF n =10). We performed glucose tolerance test (GTT) and insulin tolerance test (ITT) at the last week before organs collection. HOMA IR index was calculated. Quantitative PCR was used to determine hepatic expression of glucose-6-phospatase (G6Pase), glucokinase (GK), phosphoenolpyruvate carboxykinase (PEPCK), fructose 1,6-bisphosphatase (FBPase1) and hepatocyte nuclear factor 4 alpha (HNF4A). We also performed western blot to analyze AKT phosphorylation in liver. Two-way ANOVA was used to analyze differences between groups. Values were expressed as mean ± SEM and p ≤0.05 was considered statistically significant. Results: Although the increment of glucose area under the curve during GTT was lower in B1RKO-CAF than WT-CAF (WT-SC 46.246 ± 4.302 vs. B1RKO-SC 35.699 ± 3.383; p=0.03; and B1RKO-CAF 28.521 ± 5.338 vs. WT-CAF 56.564 ± 3.477; AUC p=0.0001), B1RKO mice presented a similar decay in glucose levels during ITT as well as HOMA-IR values . While there was a decrease in liver mRNA expression of G6Pase and PEPCK, GK expression increased in mice fed with CAF diet, without differences between genotypes. FBPase1 and HNF4A mRNA expression were not different between groups. Liver AKT phosphorylation increased with CAF in both C57Bl/6 wild type and B1RKO mice. Conclusion: The increased glucose uptake of B1RKO mice during GTT might not be related to insulin signaling in the liver, since no changes were observed in insulin resistance or in the hepatic expression of gluconeogenesis regulatory enzymes. Disclosure P. Espíndola Correia: None. C.B. Gomes: None. P.N. Merello: None. G.R. Natividade: None. C.C. Barros: None. F. Gerchman: None. Funding Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (16-0397)

Mouse Model of Non-Alcoholic Fatty Liver Disease That Replicates the Human Disease

Abstract Objectives To develop a mouse model of non-alcoholic fatty liver disease (NAFLD) that replicates the characteristic of the disease in humans, including development of obesity, dysbiosis, fibrosis and liver tumors. Methods Agouti yellow (Ay) mice, which have hyperphagia, were fed a Western diet (WD) (42% kcal from fat, 341 g/Kg sucrose, and 0.2% cholesterol) and a drinking solution containing fructose and glucose equivalent to 42 g/L of high fructose corn syrup (HFCS). Wild-type mice fed a low-fat diet (LFD) (10% kcal from fat) served as lean controls. After 16 weeks of diet treatment, tissues were collected and analyzed. Obesity was evaluated via body weight and body composition. NAFLD was evaluated by histology and confirmed by liver lipid quantification, plasma ALT and AST levels, expression of inflammation-related genes, and fibrosis-specific staining. Gene expression profile was evaluated by RNAseq. Gut microbiota dysbiosis was evaluated by 16S rRNA metagenomics. Results Ay mice fed the Western diet and HFCS for 16 weeks developed obesity and NAFLD. Histological evaluation determined that the mice developed non-alcoholic steatohepatitis (NASH) with steatosis, liver injury, inflammation, and fibrosis. This was confirmed by the following analyses: In these mice, steatosis, quantified by liver TAG content, increased 4X when compared to lean controls. Liver injury, assessed by measuring plasma ALT and AST, increased 11X and 4X, respectively. Inflammation, evaluated by liver mRNA expression of Tnf and CCl2, increased 6X and 12X, respectively. Fibrosis, quantified morphometrically, increased 2.5X. Gene expression profiling showed increases in inflammation- and fibrosis-related pathways, similar to NASH in humans. In addition, these mice developed gut microbiota dysbiosis, with increased Bacteroidetes and Proteobacteria and decreased Firmicutes, as reported in NASH in humans. Finally, 60% of the Ay mice developed liver tumors when fed the WD + HFCS diet for one year. Conclusions Ay mice fed a Western diet and HFCS developed obesity, gut microbiota dysbiosis, and NASH, including fibrosis and tumors, replicating the characteristics of NASH in humans. We present this as a new model for studying NASH physiopathology and testing new therapies. Funding Sources NIH CUNY.

Sex-dependent metabolic effects of pregestational exercise on prenatally stressed mice.

Stressful events during the prenatal period have been related to hyperactive hypothalamic-pituitary-adrenal (HPA) axis responses as well as metabolic changes in adult life. Moreover, regular exercise may contribute to the improvement of the symptoms associated with stress and stress-related chronic diseases. Therefore, this study aims to investigate the effects of exercise, before the gestation period, on the metabolic changes induced by prenatal stress in adult mice. Female Balb/c mice were divided into three groups: control (CON), prenatal restraint stress (PNS) and exercise before the gestational period plus PNS (EX+PNS). When adults, the plasmatic biochemical analysis, oxidative stress, gene expression of metabolic-related receptors and sex differences were assessed in the offspring. Prenatal stress decreased neonatal and adult body weight when compared to the pregestational exercise group. Moreover, prenatal stress was associated with reduced body weight in adult males. PNS and EX+PNS females showed decreased hepatic catalase. Pregestational exercise prevented the stress-induced cholesterol increase in females but did not prevent the liver mRNA expression reduction on the peroxisome proliferator-activated receptors (PPARs) α and γ in PNS females. Conversely, PNS and EX+PNS males showed an increased PPARα mRNA expression. In conclusion, pregestational exercise prevented some effects of prenatal stress on metabolic markers in a sex-specific manner.

<i>Coleus forskohlii</i> Extract Attenuated the Beneficial Effect of Diet-Treatment on NASH in Mouse Model

Obesity is one of the main causes of non-alcoholic steatohepatitis (NASH), which is associated with impaired liver functions including drug metabolism. Coleus forskohlii extract (CFE) is a popular ingredient of weight loss dietary supplements in Japan. In this study, we examined the effect of CFE on the treatment of NASH. C57BL/6 mice (male, 10-wk-old) were fed a NASH diet (high-fat, low-methionine, and choline-deficient diet) for 12 wk to establish NASH. Then, we examined the effect of 0.5% (w/w) CFE in diet during diet-treatment (change to control diet) and/or treadmill-exercise (45 min at 20 m/min, 5 d/wk) to improve NASH for 3 wk. After experimental period, lipids profiles and liver functional markers in the blood, and hepatic lipid content and major CYP subtype mRNA expression and activity in liver were measured. Diet-treatment, but not exercise decreased liver weight and hepatic lipid contents in NASH induced mice. CFE attenuated the effects of diet-treatment which reduced liver weight, even though body weight and adipose tissue weight were reduced. Further, CFE significantly increased liver microsomal CYP1A1, CYP1A2, CYP2C, and CYP3A activities in each condition, and CYP inductions were greater in diet-treatment group compared to those in exercise group. These results suggest that taking CFE should be avoided during diet-treatment of NASH, especially in patients under medication.

Berberine (BBR) Induced Bile Salt Export Pump (Bsep) Expression in Mouse Liver and Cultured Hepatoma Cells

Bile salt export pump (Bsep) is responsible for the biliary excretion of majority of bile salts in the liver. Dysfunction of Bsep often leads to disruption of the enterohepatic circulation of bile salts and thus cholestasis. Berberine (BBR), an active component of certain traditional herbal medicine such as Goldenseal, shows anti‐oxidative and anti‐inflammatory properties. Nuclear factor erythroid 2‐related factor (Nrf) 2 plays important roles in fighting against oxidative stress and inflammation. We recently showed that BBR increased Bsep mRNA expression in mouse liver. However, the underlying mechanism by which BBR induces Bsep expression remains largely unknown. The present study was designed to determine whether BBR induces Bsep expression via activation of Nrf2 signaling. BBR dose‐dependently increased mRNA and protein expression of Bsep in mouse liver. The studies, conducted in cultured mouse and human hepatoma cells, showed that BBR (1) increased Bsep/BSEP mRNA and protein expression, (2) enhanced the cellular efflux of Dichlorofluorescin diacetate, a known Bsep/BSEP substrate, and (3) stimulated the Bsep/BSEP gene promoter‐driven luciferase reporter activity. To determine whether Nrf2 activity is responsible for BBR‐induced Bsep expression, we knocked down Nrf2 expression in cultured hepatoma cells by using silence RNA (siRNA) that specifically targets Nrf2. Our results showed that after Nrf2 knock‐down, BBR no longer induced BSEP mRNA expression in human hepatoma cells. In contrast, in mouse hepatoma cells, Nrf2 silence partially attenuated BBR‐induced Bsep expression. In addition, promoter partial‐deletion followed by promoter‐driven luciferase reporter assay showed that deletion of putative Nrf2 binding site(s) in mouse and human Bsep/BSEP gene promoter abolished BBR‐induced luciferase activity in human but not mouse hepatoma cells. In conclusion, BBR induces human BSEP expression via Nrf2 signaling. In contrast, BBR induced Bsep expression in mouse liver or hepatoma cells independent of Nrf2 signaling.

Excessive folic acid supplementation in pregnant mice impairs insulin secretion and induces the expression of genes associated with fatty liver in their offspring

ObjectivePrevious human and animal studies have shown that excessive maternal intake of folic acid (FA) predisposes to impaired glucose tolerance in the offspring. However, the underlying mechanism is unknown. Therefore, we aimed to determine whether excessive supplementation with FA during pregnancy affects the glucose tolerance of mouse offspring. Research methods & proceduresPregnant C57BL/6J mice were fed AIN93G diet containing either 2 mg [control group (CN)] or 40 mg [high FA group (HFA)] FA/kg diet throughout their pregnancies. On postnatal days (PD)22 and 50, fasting blood glucose was measured in the offspring of both groups, and an oral glucose tolerance test (OGTT) was performed on PD50. On PD53, tissues were collected, and the tissue masses, area of insulin expression in the pancreas, liver triglyceride content, and gene expression were determined. ResultsThe blood glucose concentrations at 60 and 120 min of the OGTT were higher in female HFA than CN offspring. The serum fasting and non-fasting insulin concentrations and the area of insulin expression in the pancreas were lower in HFA than CN offspring. The liver triglyceride content was higher in female, and tended to be higher in male (P < 0.05), HFA offspring than CN offspring (P < 0.05). The liver mRNA expression of fat synthesis genes, such as Pparγ2 (male and female) and Cidec (male), was higher in HFA than CN offspring (P < 0.05). ConclusionExcessive maternal supplementation of FA in mice leads to lower insulin synthesis and an impairment in hepatic fat metabolism in the offspring.

Berbamine ameliorates ethanol-induced liver injury by inhibition of hepatic inflammation in mice.

Alcoholic liver disease (ALD) has become one of the leading causes of death in the world. Berbamine (BM), a natural product mainly derived from Berberis vulgaris L, possesses multiple bioactivities as a traditional medicine. However, the protective effect of BM on ALD remains unknown. In this study, we investigated the effect of BM on ethanol-induced hepatic injury in mice and its underlying mechanism. It was shown that BM at 0.3125-40 μmol·L-1had no effect on macrophages and hepatocytes proliferation. BM at 5-20 μmol·L-1 significantly inhibited lipopolysaccharide (LPS) or acetate-induced IL-1β and IL-6 mRNA expression in RAW264.7 cells. Moreover, BM treatment significantly inhibited LPS-induced p65 and STAT3 phosphorylation in RAW264.7 cells. Hepatic histopathology analysis showed that inflammatory cells infiltration and lipid accumulation were suppressed by 25 and 50 mg·kg-1 BM administration in ethanol-induced hepatic injury mouse model. Meanwhile, BM treatment significantly inhibited serum ALT and AST levels in ethanol-fed mice. Oil red O staining results showed that BM administration ameliorated hepatic lipid accumulation in ethanol-fed mice. Preventions of ethanol-induced hepatic injury by BM were reflected by markedly decreased serum and hepatic triglyceride (TG) and total cholesterol (TC) contents. Real-time PCR results showed that BM treatment significantly inhibited pro-inflammatory cytokines mRNA expression in ethanol-fed mouse liver. Remarkably, the mechanism of action of BM was related to the reduction of ethanol-induced NF-κB and STAT3 phosphorylation levels in liver. In addition, BM treatment significantly inhibited ERK phosphorylation but not JNK and p38 of MAPK pathway. Taken together, our results demonstrate a beneficial effect of BM on ethanol-induced liver injury via a mechanism associated with inactivation of NF-κB, STAT3 and ERK pathway, which gives insight into the further evaluation of the therapeutic potential of BM for ALD.

Antibacterial and immunoregulatory activity of a novel hepcidin homologue in diploid hybrid fish (Carassius auratus cuvieri ♀ × Carassius auratus red var ♂)

Hepcidin, a multifunctional hormone oligopeptide, not only exhibits a regulatory role in iron metabolism, but also participates in the regulation of teleostean immunity. In this study, ORF sequence of WR-hepcidin was 258 bp and encoded 85 amino acid residues. Tissue-specific analysis revealed that the highest expression of WR-hepcidin was observed in liver. Aeromonas hydrophila challenge can sharply increased WR-hepcidin mRNA expression in liver, trunk kidney and spleen. The purified WR-hepcidin fusion peptide can directly bind to A. hydrophila and Streptococcus agalactiae, reduce the relative bacterial activity, limit bacterial growth and attenuate their dissemination to tissues in vivo. In addition, the treatment of WR-hepcidin fusion protein can diminish the production of pro-inflammatory cytokines. These results indicated that WR-hepcidin can play a negative regulatory role in bacteria-stimulated pro-inflammatory cytokines production and MyD88-IRAK4 activation.

Study on the protection of water extracts of Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata against NAFLD and its mechanism

Ethnopharmacological relevancePolygoni Multiflori Radix (PMR) and Polygoni Multiflori Radix Praeparata (PMRP) that is used after processing are two well-known traditional Chinese medicines. PMRP is traditionally reported to have lipid-reducing activity as recorded in Chinese Pharmacopoeia.Aim of the study: This study aims to observe the alleviation of Polygoni Multiflori Radix Praeparata water extract (PMRPWE) and Polygoni Multiflori Radix water extract (PMRWE) against nonalcoholic fatty liver disease (NAFLD), and its potential engaged mechanism and the main active ingredients. Materials and methodsThe contents of 2,3,5,4′-tetrahydroxy-stilbene-2-O-β- D-glucoside (TSG), emodin and physcion in PMRWE and PMRPWE were measured by using high-performance liquid chromatography (HPLC). NAFLD was induced in rats by high-fat diet (HFD) feeding for 8 weeks. At the same time, rats were orally given with PMRWE (70, 140, 280 mg/kg) or PMRPWE (70, 140, 280 mg/kg) every day. Serum and liver biochemical parameters, hepatic gene expression and enzymatic activity were detected. Cellular lipids accumulation in human normal liver L-02 cells was induced by 0.5 mM non-esterified fatty acid (NEFA). ResultsThe results of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), liver reactive oxygen species (ROS) and hematoxylin-eosin (H&E) observation showed that PMRWE and PMRPWE both alleviated liver injury in HFD-fed rats. The results of liver triglyceride (TG), total cholesterol (TC) and NEFA amounts, and liver Oil Red O staining evaluation showed that PMRWE and PMRPWE both reduced hepatic lipids accumulation in HFD-fed rats. The results of 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene (BODIPY) fluorescence staining and cellular TG content showed that both PMRWE and PMRPWE reduced NEFA-induced cellular lipids accumulation in L-02 cells. PMRWE and PMRPWE increased liver mRNA expression of some signals involved in mitochondrial β oxidation, including the key enzyme carnitine palmitoyltransferase 1A (CPT1A). Moreover, PMRWE and PMRPWE increased the decreased liver CPT1A enzymatic activity in HFD-fed rats. Etomoxir (ETO), a CPT1A inhibitor, weakened the lipid-lowering activity of PMRWE and PMRPWE in vitro. Additionally, the main compounds in PMRWE and PMRPWE including TSG, emodin, physcion and resveratrol all reduced cellular lipids accumulation induced by NEFA in L-02 cells. ConclusionsPMRWE and PMRPWE alleviated NAFLD through promoting mitochondrial β oxidation by enhancing liver CPT1A activity. Stilbenes (including TSG, polydatin and resveratrol) and anthraquinones (including physcion, emodin and rhein) may be the main active compounds contributing to the lipid-lowering activity provided by PMRWE and PMRPWE.

Influence of bamboo vinegar powder supplementation on growth performance, apparent digestibility and expression of growth-related genes in finishing pigs

The study was conducted with the objective to evaluate the effect of bamboo vinegar powder (BV) on growth performance and expression of growth-related genes in finishing pigs. Ninety pigs (140-d-old, 80.88±0.82 kg) were randomly allocated to five groups and were fed on a basal diet alone (CON; served as a control), or supplemented with either antibiotics (AB) or BV at 0.5 (BV-0.5), 1.0 (BV-1.0) and 1.5 (BV-1.5) per cent levels. Relative growth parameters and nutrient apparent digestibility were measured. Expression of IGF-1, GHR, and LEP in muscle, liver, and adipose tissues were quantified by RT-q PCR. Addition of 1.0% BV significantly (P<0.05) increased ADG and reduced FCR of finishing pigs than CON. Additionally, the CP digestibility in BV-1.0 and BV-0.5 notably improved (P<0.05) than those in other groups. The pigs showed significantly (P<0.05) elevated levels of IGF-1 and GHR mRNA expression in liver in BV-1.0 and BV-1.5 than that in other groups. The IGF-1 also showed significant higher (P<0.05) expression in adipose tissues in BV-1.0 and BV-1.5 when compared to other groups. The expression of LEP increased significantly (P<0.05) in muscle and liver tissues in BV-1.0 than that in other groups. Overall, it is concluded that BV can be supplemented at 1.0% level to promote growth and development in finishing pigs.

The role of cellular prion protein in lipid metabolism in the liver

ABSTRACT Cellular prion protein (PrPC) is a plasma membrane glycophosphatidylinositol-anchored protein and it is involved in multiple functions, including neuroprotection and oxidative stress. So far, most of the PrPC functional research is done in neuronal tissue or cell lines; the role of PrPC in non-neuronal tissues such as liver is only poorly understood. To characterize the role of PrPC in the liver, a proteomics approach was applied in the liver tissue of PrPC knockout mice. The proteome analysis and biochemical validations showed an excessive fat accumulation in the liver of PrPC knockout mice with a change in mRNA expression of genes linked to lipid metabolism. In addition, the higher Bax to Bcl2 ratio, up-regulation of tgfb1 mRNA expression in PrPC knockout mice liver, further showed the evidences of metabolic disease. Over-expression of PrPC in fatty acid-treated AML12 hepatic cell line caused a reduction in excessive intracellular fat accumulation; shows association of PrPC levels and lipid metabolism. Therefore, based on observation of excessive fat globules in the liver of ageing PrPC knockout mice and the reduction of fat accumulation in AML12 cell line with PrPC over-expression, the role of PrPC in lipid metabolism is described.

Soybean oil attenuates the onset of non-alcoholic steatohepatitis and insulin resistance

AbstractBackground and Aims:General overnutrition, and a diet rich in fat and sugar are among the key risk factors for the development of metabolic diseases including diabetes type 2 and non-alcoholic fatty liver diseases (NAFLD). While being among the most commonly consumed oils world-wide, the effects of soybean oil on the development of metabolic diseases are yet not understood. Indeed, existing data is in part contradictory. Based on this background, the aim of the present study was to investigate the effects of soybean oil consumption on the development of non-alcoholic steatohepatitis (NASH), the more progressed stage of NAFLD, and insulin resistance.Methods:Female C57BL/6J mice were fed either a liquid standard control diet (C) or a liquid fat-, fructose- and cholesterol-rich diet (FFC, 25E% butterfat, 50% (wt/wt) fructose, 0.155% (wt/wt) cholesterol) or a FFC supplemented with soybean oil (FFC + S, 21E% butterfat + 4E% soy oil), with FFC and FFC + S-fed groups being pair fed for 13 weeks to induce steatohepatitis. After 7 and 12 weeks of feeding, a glucose tolerance test was performed. Indicators of liver damage, inflammation, intestinal barrier function and markers of insulin signaling cascade were measured in intestinal, liver and muscle tissue.Results:As expected FFC-fed mice developed early signs of NASH and insulin resistance. Despite similar caloric intake and body weight gain, development of NASH and insulin resistance were significantly attenuated in FFC + S-fed mice when compared to FFC-fed animals. Indeed, signs of hepatic inflammation e.g. number of inflammatory foci and neutrophils as well as activity of transaminase in plasma were almost at the level of control in FFC + S-fed mice. Prevalence of macrovesicular steatosis being the predominant type of fat accumulation found in FFC-fed animals was also markedly lower in FFC + S-fed mice. Furthermore, while in muscle and liver tissue of FFC-fed mice insulin receptors mRNA expression was significantly different from C-fed animals, insulin receptors expression in FFC + S-fed mice was almost at the level of controls. The protective effects of soybean oil supplementation on the development of metabolic alterations were associated with a protection against the induction of TLR4 mRNA expression and dependent signaling molecules in liver tissue.Conclusion:Taking together, our results indicate that supplementation with soybean oil may attenuate the development of diet-induced NASH and insulin resistance in mice and that this may be related to alteration of TLR4-dependent signaling cascade in liver tissue. (Funded in parts by UFOP e.V.)

Maternal Dietary Fiber Composition during Gestation Induces Changes in Offspring Antioxidative Capacity, Inflammatory Response, and Gut Microbiota in a Sow Model.

To study the effects of maternal dietary fiber composition during gestation on offspring antioxidant capacity, inflammation, and gut microbiota composition, we randomly assigned 64 gilts to four treatments and administered diets with an insoluble/soluble fiber ratio of 3.89 (R1), 5.59 (R2), 9.12 (R3), and 12.81 (R4). Sow samples (blood and feces at gestation 110) and neonatal samples (blood, liver, and colonic contents) were collected. The results showed that sows and piglets in R1 and R2 had higher antioxidant enzyme activity and lower pro-inflammatory factor levels than those in R3 and R4. Moreover, piglets in R1 and R2 had higher liver mRNA expression of Nrf2 and HO-1 and lower NF-κB than piglets in R4. Interestingly, maternal fiber composition not only affected the production of short-chain fatty acids (SCFAs) in sow feces but also influenced the concentrations of SCFAs in the neonatal colon. Results of high-throughput sequencing showed that piglets as well as sows in R1 and R2 had microbial community structures distinct from those in R3 and R4. Therefore, the composition of dietary fiber in pregnancy diet had an important role in improving antioxidant capacity and decreasing inflammatory response of mothers and their offspring through modulating the composition of gut microbiota.

Lactobacillus plantarum KSFY06 on d-galactose-induced oxidation and aging in Kunming mice.

Yogurt from Xinjiang, China, is a traditional Chinese fermented food rich in beneficial microorganisms, such as Lactobacillus plantarum KSFY06. In this study, the effect of KSFY06 on oxidative aging was investigated using live animal experiments. Molecular biological methods were used to analyze the serum and tissues of mice with oxidative aging induced by d‐galactose, which showed that KSFY06 can inhibit the decline of heart, liver, spleen, and kidney caused by aging. The KSFY06 strain increased the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px), catalase (CAT), and glutathione (GSH) in serum and liver of aging mice, while the content of malondialdehyde (MDA) is reduced. Pathological observation showed that KSFY06 alleviated damage to the liver, spleen, and skin of oxidative aging mice. qPCR showed that, at high dose (2 × 109 cfu/kg per day), KSFY06 upregulates copper/zinc superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), catalase (CAT) mRNA expression, and its downstream inducible nitric oxide synthase (iNOS) mRNA expression in liver and spleen tissues induced by d‐gal. To a certain extent, these findings indicate that L. plantarum KSFY06 is able to protect against oxidative stress in the d‐gal‐induced aging model. In conclusion, L. plantarum KSFY06 may provide a potential research value in the prevention or alleviation of related diseases caused by oxidative stress.

Changes in meta-transcriptome of rumen epimural microbial community and liver transcriptome in young calves with feed induced acidosis

The common management practices of dairy calves leads to increased starch concentration in feed, which subsequently may cause rumen acidosis while on milk and during weaning. Until recently, few attempts were undertaken to understand the health risks of prolonged ruminal acidosis in post weaning calves. Resultantly, the molecular changes in the digestive tracts in post-weaning calves with ruminal acidosis remain largely unexplored. In this study, we investigated the liver transcriptome changes along with its correlation with the rumen microbial rRNA expression changes in young calves using our model of feed induced ruminal acidosis. In this model, new born calves were fed a highly processed, starch-rich diet starting from one week of age through 16 weeks. A total of eight calves were involved in this study. Four of them were fed the acidosis-inducing diet (Treated) and the rest of the four were fed a standard starter diet (Control). Liver and rumen epithelial tissues were collected at necropsy at 17 weeks of age. Transcriptome analyses were carried out in the liver tissues and rRNA meta-transcriptome analysis were done using the rumen epithelial tissues. The correlation analysis was performed by comparing the liver mRNA expression with the rumen epithelial rRNA abundance at genus level. Calves with induced ruminal acidosis had significantly lower ruminal pH in comparison to the control group, in addition to significantly less weight-gain over the course of the experiment. In liver tissues, a total of 428 differentially expressed genes (DEGs) (fold-change, FC ≥ 1.5; adjusted P ≤ 0.1) were identified in treated group in comparison to control. Biological pathways enriched by these DEGs included cellular component organization, indicating the impact of ruminal acidosis on liver development in young calves. Specifically, the up-regulated genes were enriched in acute phase response (P < 0.01), pyruvate metabolic process (P < 0.01) and proton-acceptors (P ≪ 0.001), indicating the liver’s response to feed induced acidosis at the transcriptome level. Twelve transferase activity related genes had significant correlation with rumen microbial rRNA expression changes. Among these genes, two up-regulated genes were reported with involvement in lipid metabolism in the liver, implying the direct effect of feed-induced acidosis on both the rumen microbial community and liver metabolism. Our study provides insight into the physiological remodeling in the liver resultant from the prolonged acidosis in post weaning calves, which may facilitate future RNA-seq based diagnosis and precision management of rumen acidosis in dairy calves.

Tibial growth plate vascularization is inhibited by the dithiocarbamate pesticide thiram in chickens: potential relationship to peripheral platelet counts alteration.

The widespread use of thiram has raised concerns for health and its toxic effects, but the underlying toxicity mechanism on platelets and bones is poorly defined. Here, we found a significant increase in the number of platelets in chickens with the thiram intake, due to the increased expression of thrombopoietin mRNA in the dysfunction liver. Furthermore, the decreased vascular distribution and cell death of chondrocytes in the tibial growth plates (TGPs) were observed, resulting in bone growth inhibition, which is associated with the abnormal activation of platelets leading to the extraordinary decrease of vascular endothelial growth factor A (VEGFA) and angiopoietin-1 protein were released and their corresponding receptors VEGFR2 and Tie-2 expressions were also reduced in the TGPs. Taken together, these findings revealed that thiram has an adverse effect on bones and platelets, which may have a high risk of thrombosis and osteoarthritis.

A novel LEAP-2 in diploid hybrid fish (Carassius auratus cuvieri ♀ × Carassius auratus red var. ♂) confers protection against bacteria-stimulated inflammatory response

LEAP-2, a multifunctional peptide, not only exhibits a regulatory role in pathogenic infection, but also participates in the regulation of teleostean immunity. In this study, ORF sequence of WR-LEAP-2 was 240 bp and encoded 79 amino acid residues. Tissue-specific analysis revealed that the highest expression of WR-LEAP-2 was observed in liver. Aeromonas hydrophila challenge can sharply increase WR-LEAP-2 mRNA expression in liver, kidney and spleen. The purified WR-LEAP-2 peptide can directly bind to A. hydrophila and S. agalactiae, reduce the relative bacterial activity and limit bacterial growth in vitro. In addition, the treatment of WR-LEAP-2 can restrict bacterial dissemination in vivo and reduce production of pro-inflammatory cytokines. These results indicated that WR-LEAP-2 can confer protection against A. hydrophila- or S. agalactiae-stimulated MyD88-dependent pro-inflammatory cytokines activation.

The potential of antioxidant-rich Maoberry (Antidesma bunius) extract on fat metabolism in liver tissues of rats fed a high-fat diet

BackgoundObesity and dyslipidemia are major risk factors associated with non-alcoholic fatty liver disease (NAFLD). NAFLD refers to the accumulation of fat in more than 5% of the liver without alcohol consumption. NAFLD is the most common liver disease and is rapidly becoming a global public health problem. Maoberry (Antidesma bunius) is a fruit rich in antioxidants, especially phenolic compounds, which are reported to have benefits for patients with NAFLD.MethodsWe evaluated the effect of Maoberry extract on fat metabolism in liver tissues of high fat diet–induced rats. Five (5) groups (n = 12) of male Sprague-Dawley (SD) rats were divided into those given a high fat diet with no treatment (HF), different dosages of Maoberry extracts (0.38 [ML], 0.76 [MM) and 1.52 [MH] g/kg body weight) and 10 mg/kg statin (STAT). The rats were fed a high fat diet for 4 weeks to induce obesity and subsequently continued more for 12 weeks with treatments of Maoberry extracts or STAT. The levels of triglyceride, liver enzymes, oxidative stress and inflammation markers, triglyceride synthesis regulators, and pathology of the liver in high fat diet-induced rats were investigated.ResultsFeeding Maoberry extract in MH groups resulted in decreasing levels of serum alanine aminotransferase (ALT), liver triglyceride, liver thiobarbituric acid reactive substances (TBARS) and mRNA expression of tumour necrosis factor (TNF)-α, interleukin (IL)-6, glycerol-3-phosphate acyltransferase (GPAT)-1 and acetyl-coenzyme A carboxylase (ACC) compared with the HF group (P < 0.05). Moreover, histopathological study of the liver showed reduced fat droplets in the Maoberry extract treatment groups, especially in MH groups and STAT treatment groups.ConclusionsThe improvements of fat metabolism in liver tissues of rats fed a high-fat diet were observed in Maoberry extracts treatment groups. The underline mechanism that link to fat metabolism might be through the process accompanied with down-regulated the gene expression of key enzymes of lipid production, antioxidant activity, and anti-inflammation properties of Maoberry extracts which contains high levels of phenolic and flavonoid compounds.