Zhiqing Huang

Biological System Responses to Zearalenone Mycotoxin Exposure by Integrated Metabolomic Studies

This study aims to investigate the effect of zearalenone supplementation on rat metabolism. Rats received biweekly intragastric administration of zearalenone mycotoxin (3 mg/kg body weight) for 2 weeks. Urine and plasma samples after zearalenone administration were analyzed by NMR-based metabolomics. Zearalenone exposure significantly elevated the plasma levels of glucose, lactate, N-acetyl glycoprotein, O-acetyl glycoprotein, and propionate but reduced the plasma levels of tyrosine, branched-chain amino acids, and choline metabolites. Zearalenone supplementation decreased the urine levels of butyrate, lactate, and nicotinate. However, it increased the urine levels of allantoin, choline, and N-methylnicotinamide at 0-8 h after the last zearalenone administration and those of 1-methylhistidine, acetoacetate, acetone, and indoxyl sulfate at 8-24 h after the last zearalenone administration. These results suggest that zearalenone exposure can cause oxidative stress and change common systemic metabolic processes, including cell membrane metabolism, protein biosynthesis, glycolysis, and gut microbiota metabolism.

Metabolomic Strategy for the Detection of Metabolic Effects of Spermine Supplementation in Weaned Rats

The purpose of this study is to examine the effects of spermine supplementation on weaned rat metabolism. A metabolomic strategy employing high-resolution (1)H NMR spectroscopy and multivariate data analysis was used to investigate rat biological responses to spermine ingestion. Rats received intragastric administration of either 0.2 or 0.4 μmol/g body weight of spermine or saline for 3 days. Plasma samples taken 48 h after the last spermine ingestion were analyzed. Spermine supplementation significantly increased the plasma levels of 1-methylhistidine, 3-hydroxybutyrate, alanine, glutamate, glycerolphosphocholine, phosphorylcholine, myo-inositol, phenylalanine, lysine, glutamine, trimethylanine, tyrosine, valine, formate, glucose, and lipids. These results suggest that spermine ingestion can alter common systemic metabolic processes, including cell membrane metabolism, lipid metabolism, glucose-alanine cycle metabolism, amino acid metabolism, and gut microbiota metabolism. This study also shows the important role of spermine administration in modulating the metabolism of weaned rats.

Role of microRNA-27a in myoblast differentiation.

MicroRNAs (miRNAs) are a class of endogenous non‐coding RNAs that are critically involved in roles in various aspects of skeletal myogenesis. microRNA miR‐27a promotes myoblast proliferation by targeting myostatin, a critical inhibitor of skeletal muscle development, but its mode of action in myoblast differentiation remains unclear. We have found that expression of miR‐27a and myostatin were upregulated and downregulated, respectively, during myoblast differentiation. Overexpression of miR‐27a increased the number of myosin heavy chain (MHC)‐positive cells and upregulated mRNA and protein of MyoD and myogenin. These findings indicate that miR‐27a plays a role in enhancing myoblast differentiation.

NMR-based metabolomic studies reveal changes in biochemical profile of urine and plasma from rats fed with sweet potato fiber or sweet potato residue

Dietary fiber has attracted more interest in recent years because many studies have uncovered its disease preventive and health-promoting features: containing blood cholesterol and/or glucose attenuation and reducing obesity risk. However, the health effects of sweet potato fiber (SF) and sweet potato residue (SR) and the knowledge of their exact mechanisms of action are still not fully understood. This study investigates the effect of SF and SR administration on rat metabolism. Rats were randomly assigned to one of three dietary groups of 11 rats each and given a basal diet containing 15% SF, 15% SR, or no supplemental fiber (control). The groups were observed for 30 days, and urine and plasma samples were analyzed by proton nuclear magnetic resonance. Compared with the rats in the control group, those administered the diet containing SF exhibited significantly increased plasma levels of lipid, lactate, and myo-inositol, as well as urine levels of acetate, citrulline, N-acetylglutamate, and p-hydroxyphenylacetate. SF significantly decreased the plasma levels of glutamine, glutamine/glutamate, lysine, phosphorylcholine/glycerolphosphocholine, tyrosine, and glucose, as well as the urine level of allantoin. Moreover, SR significantly increased the plasma levels of acetone, lipid, and lactate, as well as the urine levels of acetamide, alanine, citrulline, ethanol, lactate, methylamine, methylmalonate, N-acetylglutamate, and α-hydroxybutyrate, compared with the control group. SR significantly decreased the plasma levels of citrate, glutamate, glutamine, isoleucine, lysine, methionine, and glucose, as well as the urine level of isobutyrate. These results suggest that SF and SR supplementation have certain systemic metabolic processes in common, including lipid metabolism, glycogenolysis and glycolysis metabolism, energy metabolism, protein biosynthesis, and gut microbiota metabolism. This study demonstrates the potential for the routine use of metabolomics in nutritional studies to characterize metabolic effects and understand the influence of diet on animal metabotypes.

Tissue Distribution of Porcine FTO and Its Effect on Porcine Intramuscular Preadipocytes Proliferation and Differentiation.

The fat mass and obesity associated (FTO) gene plays an important role in adipogenesis. However, its function during porcine intramuscular preadipocyte proliferation and differentiation remains poorly understood. In this study, we prepared the antiserum against porcine FTO (pFTO), which was used to determine its subcellular localization and tissue distribution. Our data indicated that pFTO was localized predominantly in the nucleus. Real-time quantitative PCR and western blot analysis showed that pFTO was highly expressed in the lung and subcutaneous adipose tissue. Overexpression of pFTO in porcine intramuscular preadipocytes significantly promoted cell proliferation and lipid deposition. Furthermore, overexpression of pFTO in differentiating porcine intramuscular preadipocytes also significantly increased the mRNA levels of adipocyte differentiation transcription factors peroxisome proliferators-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), lipoprotein lipase (LPL) and fatty acid synthase (FAS). Our findings provide the first functional evidence to reveal a role of pFTO in porcine intramuscular preadipocyte proliferation and differentiation.

Effect of 25-hydroxyvitamin D3 on rotavirus replication and gene expressions of RIG-I signalling molecule in porcine rotavirus-infected IPEC-J2 cells.

The study evaluated whether a 25-hydroxyvitamin D3 (25D3) supplementation decreases the replication of rotavirus by the retinoic acid-inducible gene I (RIG-I) signalling pathway in a porcine small intestinal epithelial cell line (IPEC-J2). The results show that IPEC-J2 cells express high baseline levels of 1α-hydroxylase (CYP27B1), which converts inactive 25D3 to the active 1,25-dihydroxyvitamin D3 (1,25D3). Porcine rotavirus (PRV) infection alone resulted in a significant increase in CYP27B1 mRNA, which augmented the production of active vitamin D. Physiological concentrations of 25D3 were found to decrease PRV replication in IPEC-J2 cells. RIG-I plays an important role in the recognition of double-stranded RNA virus by host cells. Upon recognition, RIG-I triggers a series of signalling molecules such as interferon-β (IFN-β) promoter stimulator 1 (IPS-1) leading to the expression of type I interferons (IFN-β). Active 25D3 that was generated by PRV-infected IPEC-J2 cells led to an increased expression of toll-like receptors 3 (TLR3), RIG-I, IPS-1, IFN-β and IFN-stimulated genes 15 (ISG15) with important innate immune functions. Inhibiting CYP27B1 also failed to increase RIG-I, IPS-1, IFN-β and ISG15 mRNA expression. These observations suggest that 25D3 can directly inhibit PRV in IPEC-J2 cells, which requires this active form of vitamin D. The anti-rotavirus effect of 25D3 is mediated at least in part by RIG-I signalling pathways in IPEC-J2 cells.

Trace Mineral Overload Induced Hepatic Oxidative Damage and Apoptosis in Pigs with Long-Term High-Level Dietary Mineral Exposure

The present study investigated the effects of dietary trace mineral (Cu, Fe, Mn, and Zn) supplemental strategies on liver oxidative stress, endoplasmic reticulum stress, inflammation, and apoptosis of pigs. A total of 96 Duroc × Landrace × Yorkshire (DLY) piglets were randomly divided into four groups: considered or not considered the trace mineral concentrations in basal diet, and then added to the requirements proposed by NRC (2012) (+B/NR or -B/NR); and considered or not considered the basal diets trace mineral concentrations and then added to the level of commercial trace mineral supplement (+B/PL or -B/PL). Pigs were fed from 6.5 to 115 kg. Compared with +B/NR diets, -B/PL diets increased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations (P < 0.05), resulted in high levels of Fe, Cu, Mn, and Zn accumulation in liver (P < 0.05), as well as led to hepatic oxidative damage with the high concentrations of thiobarbituric acid reactive substance (TBARS), protein carbonylation (PCO), and 8-hydroxyguanine (8-OHG) in liver (P < 0.05). Furthermore, pigs fed -B/PL diets increased CCAAT/enhancer-binding protein homologous protein (CHOP), eukaryotic initiation factor-2α (eIF-2a), interleukin-6(IL-6), B-cell lymphoma leukemia-2-associated X protein (Bax), and caspase-3, caspase-8, and caspase-9 gene expression (P < 0.05) in liver. -B/PL diets also up-regulated hepatic mRNA expression of phosphoenolpyruvate carboxykinase1 (PEPCK1), glucose-6-phosphatase (G6PC), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS) (P < 0.05) and down-regulated hormone-sensitive lipase (HSL) mRNA expression (P < 0.05) when compared with those of the + B/NR diet group. Taken together, the results indicated that long-term dietary mineral exposure with the commercial supplement level could cause harm to the structure and metabolic function of liver in pigs.

Role of Phosphotyrosine Interaction Domain Containing 1 in Porcine Intramuscular Preadipocyte Proliferation and Differentiation

ABSTRACT Phosphotyrosine interaction domain containing 1 (PID1), a recently identified gene involved in obesity-associated insulin resistance, plays an important role in fat deposition. However, its effect on porcine intramuscular preadipocyte proliferation and differentiation remains poorly understood. In this study, the plasmid pcDNA3.1(+)-pPID1 was transfected into porcine intramuscular preadipocytes with Lipofectamine 3000 reagent to over-express porcine PID1 (pPID1). Over-expression of pPID1 significantly promoted porcine intramuscular preadipocyte proliferation. Expression of pPID1 mRNA was significantly increased upon porcine intramuscular preadipocyte differentiation. Indirect fluorescent immunocytochemistry demonstrated that pPID1 protein was localized predominantly in the nucleus of porcine intramuscular preadipocyte. The mRNA levels of peroxisome proliferators-activated receptor γ, CCAAT/enhancer binding protein α and lipoprotein lipase were significantly increased by pPID1 over-expression. Over-expression of pPID1 also led to an increase in lipid accumulation which was detected by Oil Red O staining, and significantly increased the intramuscular triacylglycerol content. These results indicate that pPID1 may play a role in enhancing porcine intramuscular preadipocyte proliferation and differentiation.

Potential Risk of Isoflavones: Toxicological Study of Daidzein Supplementation in Piglets

As a main component of soy isoflavones, daidzein is rich in soy-derived products, which are widely used as feed ingredients in farm animals. However, little research has been conducted on the side effects of dietary daidzein, especially in young animals. In this study, the safety of daidzein was evaluated. Results show that ingesting 400 mg/kg of dietary daidzein for 70 days is associated with a lower average daily weight gain (kilogram) (0.47 ± 0.03 vs 0.54 ± 0.04, P < 0.05) and a higher splenic damage index (1.00 ± 1.10 vs 0.00 ± 0.00, P < 0.05) in young pigs compared with control. Female pigs receiving 200 and 400 mg/kg daidzein showed reduced serum testosterone levels (ng/L) on days 35 and 70 compared with the control group (day 35, 246 ± 74 and 224 ± 20 vs 362 ± 48, P < 0.05; day 70, 252 ± 38 and 219 ± 77 vs 374 ± 38, P < 0.05). Daidzein residue (μg/kg) in pig livers increased (243 ± 80 vs 142 ± 47, P < 0.05, day 70). These results suggest that dietary supplements of 400 mg/kg of daidzein negatively affect the weight gain and splenic morphology of pigs.

Fungi in Gastrointestinal Tracts of Human and Mice: from Community to Functions

Fungi are often ignored in studies on gut microbes because of their low level of presence (making up only 0.1% of the total microorganisms) in the gastrointestinal tract (GIT) of monogastric animals. Recent studies using novel technologies such as next generation sequencing have expanded our understanding on the importance of intestinal fungi in humans and animals. Here, we provide a comprehensive review on the fungal community, the so-called mycobiome, and their functions from recent studies in humans and mice. In the GIT of humans, fungi belonging to the phyla Ascomycota, Basidiomycota and Chytridiomycota are predominant. The murine intestines harbor a more diverse assemblage of fungi. Diet is one of the major factors influencing colonization of fungi in the GIT. Presence of the genus Candida is positively associated with dietary carbohydrates, but are negatively correlated with dietary amino acids, proteins, and fatty acids. However, the relationship between diet and the fungal community (and functions), as well as the underlying mechanisms remains unclear. Dysbiosis of intestinal fungi can cause invasive infections and inflammatory bowel diseases (IBD). However, it is not clear whether dysbiosis of the mycobiome is a cause, or a result of IBD. Compared to non-inflamed intestinal mucosa, the abundance and diversity of fungi is significantly increased in the inflamed mucosa. The commonly observed commensal fungal species Candida albicans might contribute to occurrence and development of IBD. Limited studies show that Candida albicans might interact with immune cells of the host intestines through the pathways associated with Dectin-1, Toll-like receptor 2 (TLR2), and TLR4. This review is expected to provide new thoughts for future studies on intestinal fungi and for new therapies to fungal infections in the GIT of human and animals.