Isolation and Identification of Anti-Inflammatory Peptide from Goose Blood Hydrolysate to Ameliorate LPS-Mediated Inflammation and Oxidative Stress in RAW264.7 Macrophages.

This study was designed to isolate an anti-inflammatory activity oligopeptide from goose blood (GBP) for ameliorating LPS-mediated inflammation response and oxidative stress in RAW264.7 macrophages. In this study, GBP was isolated by tangential flow ultrafiltration system (TFUS) combined with size exclusion chromatography (SEC), ion exchange chromatography (IEC), and reversed-phase liquid chromatography (RP-LC), and then identified by liquid chromatography mass spectrometry (LC-MS/MS). The experiment results indicated that the amino acid sequence of oligopeptide with the best anti-inflammatory activity was IIe-Val-Tyr-Pro-Trp-Thr-Gln-Arg (IVYPWTQR), which had a molecular weight of 1062.5720 Da, and was derived from haemoglobin subunit beta OS in goose blood. In addition, IVYPWTQR was confirmed to have satisfactory stability and maintained high anti-inflammatory activity in a simulated gastrointestinal digestion. The mechanism by which the IVYPWTQR protected against LPS-mediated inflammation response was attributed to downregulating the TLR4/NF-kB/iNOS pathway. Moreover, IVYPWTQR ameliorated oxidative stress damage in inflammatory state was attributed to activating antioxidant defence system, which was regulated by Keap-1/NRF2/HO-1 signalling pathway for decreasing the accumulation of reactive oxide species (ROS). In summary, these results indicated GBP could serve as a potential functional factor for prevention and improvement of inflammation mediated by LPS and provided an affordable dietary intervention strategy to prevent inflammation.

Isolation and Identification of Antioxidative Peptide from Goose Liver Hydrolysate to Ameliorate Alcohol-Mediated Oxidative Stress Damage in HHL-5 Hepatocytes.

The aim of this study was to isolate and identify antioxidative peptide from goose liver hydrolysate (GLHP) for ameliorating oxidative stress damage by alcohol in HHL-5 hepatocytes. In this research, the target antioxidative peptides in GLHP were separated, purified, and identified via a tangential flow ultrafiltration system combined with size exclusion chromatography (SEC), ion exchange chromatography (IEC), reversed-phase liquid chromatography (RP-LC), and LC-MS/MS. The results suggested that the amino acid sequence of the target antioxidative peptide for ameliorating alcohol-mediated oxidative stress damage in HHL-5 hepatocytes was Leu-Pro-Leu-Pro-Phe-Pro (LPLPFP), which had a molecular weight of 683.41 Da, and was derived from NADH-ubiquinone oxidoreductase chain 1 in goose liver. In addition, LPLPFP was confirmed to have a satisfactory stability and maintained high hepatic protective activity in a simulated gastrointestinal digestion. Moreover, the mechanism of LPLPFP prevented against oxidative stress damage in HHL-5 hepatocytes was attributed to inhibiting the production of reactive oxide species (ROS) by upregulating genes expression in the Ahr-NQO1 signal pathway. In conclusion, these results indicated that dietary GLHP supplementation could ameliorate alcohol-mediated oxidative stress damage and provide an affordable dietary intervention strategy to prevent alcohol-mediated hepatocyte damage.

Consuming egg yolk decreases body weight and increases serum HDL and brain expression of TrkB in male SD rats.

BACKGROUND: Egg yolks contain large amounts of cholesterol and are suspected to be harmful after long-term consumption. In this experiment, 63 rats were used to evaluate the effect of egg white (EW) and egg yolk (EY) supplementation on serum lipids and brain cognition. The feeding time lasted 4 weeks after a 1-week acclimation. RESULTS: Body weight was significantly higher in rats fed 132.0 g kg-1 EW and significantly lower when fed 40 g kg-1 EY (P < 0.05). Total cholesterol and low-density lipoprotein increased in rats fed 72.0 g kg-1 EW compared with rats from NC and EY groups (P < 0.05). High-density lipoprotein (HDL) was higher in rats fed 40 g kg-1 EY and decreased when fed 72.0 g kg-1 EW (P < 0.05). Rats fed a diet with EY exhibited abundant neurons in the CA1 hippocampus and complete subcellular structures. Rats fed 132 g kg-1 EW exhibited shrunken cells and swollen mitochondria. Brain-derived neurotrophic factor had constitutively low expression among groups, while tyrosine kinase B (TrkB) exhibited higher expression levels in rats fed a diet containing EY compared with other groups (P < 0.05). CONCLUSION: EY consumption reduced body weight and increased HDL levels. Diet containing EY could improve cognition through enhanced trkB expression. © 2019 Society of Chemical Industry.

Expression of Oocyte Vitellogenesis Receptor Was Regulated by C/EBPα in Developing Follicle of Wanxi White Goose.

Yolk precursor was synthesized under regulation of hormone secretion, while the mechanism of its incorporation into follicle is still unknown. The reproductive hormones, oocyte vitellogenesis receptor (OVR) expression at pre-, early-, peak- and ceased-laying period, and localization of Wanxi White goose were determined in this study. The results showed that the concentration of LH was lowest in serum at peak laying period compared to the other periods (p < 0.01). Moreover, the concentration of E2 was highest (p < 0.01) in serum at early laying period than that of other periods. Moreover, the gene expression level of OVR was highest at ceased laying period compared to other periods (p = 0.014) and was higher in developing follicles than other follicles (p < 0.01). The OVR was distributed in the granular cell layer and decreased with the maturation of follicles. Five transcription factors were predicted in the promoter of OVR, then were screened and verified by overexpression in granulosa cells. C/EBPα and MF3 significantly stimulated the expression of OVR. The combined overexpression of C/EBPα and OVR significantly stimulated the transportation of lipid from culture medium to cytoplasm. In conclusion, C/EBPα is the key transcription factor promoting OVR expression in goose follicle granulosa cells.

Oligopeptides from Jinhua ham prevent alcohol-induced liver damage by regulating intestinal homeostasis and oxidative stress in mice.

The current study aimed to evaluate the protective activity of peptides isolated from Jinhua ham (JHP) against alcoholic liver disease (ALD) and the mechanisms by which JHP prevents against ALD. The tangential flow filtration (TFF) combined with size exclusion chromatography (SEC) and reversed-phase high performance liquid chromatography (RP-HPLC) were used to isolate the JHP. Then the hepatoprotective activity of peptides was evaluated through experiments in mice. The primary structure of the peptide with the strongest liver protective activity was Lys-Arg-Gln-Lys-Tyr-Asp (KRQKYD) and the peptide was derived from the myosin of Jinhua ham, which were both identified by LC-MS/MS. Furthermore, the mechanism of KRQKYD prevention against ALD was attributed to the fact that KRQKYD increases the abundance of Akkermansia muciniphila in the gut and decreases the abundance of Proteobacteria (especially Escherichia_Shigella). The LPS-mediated liver inflammatory cascade was reduced by protecting the intestinal barrier, increasing the tight connection of intestinal epithelial cells and reducing the level of LPS in the portal venous circulation. KRQKYD could inhibit the production of ROS by upregulating the expression of the NRF2/HO-1 antioxidant defense system and by reducing oxidative stress injury in liver cells. This study can provide a theoretical foundation for the application of JHP in the protection of liver from ALD.

A new method for assessing variability of 24 h blood pressure and its first application in 1526 elderly men.

1. Blood pressure variability (BPV) includes physiological and random variations in blood pressure (BP). Commonly used approaches, such as standard deviation (SD) and weighted standard deviation (wSD) methods, do not efficiently assess random variation in BP. In the present study, we propose a novel method to assess individual BP variations, extracting random variation in BP by eliminating physiological variation mathematically. This novel assessment method furthers our understanding of the relationship between BP variation and lacunar infarction (LACI). 2. In the present study, we analysed ambulatory blood pressure monitoring recordings taken from 1526 men aged 60-98 years of age. Individual curves were created using a mathematical method and the related BP variation calculated, namely the SD for individual BP variations. In addition, correlations between LACI and BP variations as determined by the classical SD method, wSD and our novel assessment method (SD') were evaluated. 3. The results demonstrated that 24 h variations in systolic BP (SBP) were closely associated with LACI when the SD and wSD methods were used (P < 0.05), but the most significant correlations were observed when the SD' method was used (P < 0.01). Furthermore, using SD' yielded the lowest value of the parameter P among the three different methods used to analyse BPV. Using the SD' method, a significant correlation was found between variations in SBP and the incidence of LACI (P < 0.05). It was found that the incidence of LACI increased by 2% with each 1 mmHg increase in SBP variation. 4. In conclusion, our novel assessment method enables mathematical removal of interference from physiological BP variation and the results show a better correlation with LACI. Thus, our novel method may be considered a simple index of 24 h BP variation that is superior to conventional SD and wSD methods.

Genetic Parameters for Yolk Cholesterol and Transcriptional Evidence Indicate a Role of Lipoprotein Lipase in the Cholesterol Metabolism of the Chinese Wenchang Chicken.

The yolk cholesterol has been reported to affect egg quality and breeding performance in chickens. However, the genetic parameters and molecular mechanisms regulating yolk cholesterol remain largely unknown. Here, we used the Wenchang chicken, a Chinese indigenous breed with a complete pedigree, as an experimental model, and we examined 24 sire families (24 males and 240 females) and their 362 daughters. First, egg quality and yolk cholesterol content were determined in 40-week-old chickens of two consecutive generations, and the heritability of these parameters was analyzed using the half-sib correlation method. Among first-generation individuals, the egg weight, egg shape index, shell strength, shell thickness, yolk weight, egg white height, Haugh unit, and cholesterol content were 45.36 ± 4.44 g, 0.81 ± 0.12, 3.07 ± 0.92 kg/cm2, 0.340 ± 0.032 mm, 15.57 ± 1.64 g, 3.36 ± 1.15 mm, 58.70 ± 12.33, and 274.3 ± 36.73 mg/egg, respectively. When these indexes were compared to those of the following generation, no statistically significant difference was detected. Although yolk cholesterol content was not associated with egg quality in females, an increase in yolk cholesterol content was correlated with increased yolk weight and albumin height in sire families (p < 0.05). Moreover, the heritability estimates for the yolk cholesterol content were 0.328 and 0.530 in female and sire families, respectively. Therefore, the yolk cholesterol content was more strongly associated with the sire family. Next, chickens with low and high yolk cholesterol contents were selected for follicular membrane collection. Total RNA was extracted from these samples and used as a template for transcriptional sequencing. In total, 375 down- and 578 upregulated genes were identified by comparing the RNA sequencing data of chickens with high and low yolk cholesterol contents. Furthermore, Gene Ontology term and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses indicated the involvement of energy metabolism and immune-related pathways in yolk cholesterol deposition. Several genes participating in the regulation of the yolk cholesterol content were located on the sex chromosome Z, among which lipoprotein lipase (LPL) was associated with the peroxisome proliferator-activated receptor signaling pathway and the Gene Ontology term cellular component. Collectively, our data suggested that the ovarian steroidogenesis pathway and the downregulation of LPL played critical roles in the regulation of yolk cholesterol content.

Green forage and fattening duration differentially modulate cecal microbiome of Wanxi white geese.

Gut microbial ecology is responsible for fatty acid metabolism in ruminants. The cecal microbiota composition of geese and their adaptation to fiber inclusion and feeding timeswere investigated in this study. A total of 116 Wanxi white geese were randomly selected at 70 days old. Eight geese were subjected to cecal sampling at 70 d of age, and the remaining 108 geese were divided into four groups with three replicates each (9 geese in each replicate). The geese in the four groups were fed 0, 15, 30, and 45% green forage (relative to dry matter), respectively. Three birds from each replicate were selected for cecal sampling at 80, 90, and 100 days old. All samples were subjected to 16S rRNA gene sequencing using the Illumina Ion Personal Genome Machine platform. Bacterial abundance was analyzed using two-way ANOVA analysis, and the relationship between the relative abundance of bacteria (phylum level) and fatty acids was analyzed using acanonical correspondence analysis. Cecal microbiota in geese were mainly composed of Bacteroidetes (68.46%), Firmicutes (20.04%), and Proteobacteria (7.89%). Dietary treatments had no significant effect on the α-diversity indices of the cecal bacterial community (P > 0.05), but a numerical increase occurred with increased fattening duration and green forage inclusion. The Selenomonadales order (P = 0.024), Negativicutes class (P = 0.026), and Megamonas (P = 0.012) and Oscillospira (P = 0.042) genera were affected by green forageinclusion level, and microflora abundance was mainly influenced by the fattening duration. Bacteria phyla were mostly set along the line of linolenic acid and oleic acid. Finally, Bacteroidales might be an intestinal promoter that improves unsaturated fatty acid synthesis in geese.