Riboflavin is an essential micronutrient and a precursor of flavin mononucleotide and flavin adenine dinucleotide for maintaining cell homeostasis. Riboflavin deficiency (RD) induces cell apoptosis. Endoplasmic reticulum (ER) stress is considered to induce apoptosis, and C/EBP homologous protein (CHOP) is a key pathway involved in this process. However, whether RD-induced apoptosis is mediated by ER stress and the CHOP pathway remains unclear and needs further investigation. Therefore, the current study presents the effect of RD on ER stress and apoptosis in the human hepatoma cell line (HepG2). Firstly, cells were cultured in a RD medium (4.55 nM riboflavin) and a control (CON) medium (1005 nM riboflavin). We conducted an observation of cell microstructure characterization and determining apoptosis. Subsequently, 4-phenyl butyric acid (4-PBA), an ER stress inhibitor, was used in HepG2 cells to investigate the role of ER stress in RD-induced apoptosis. Finally, CHOP siRNA was transfected into HepG2 cells to validate whether RD triggered ER stress-mediated apoptosis by the CHOP pathway. The results show that RD inhibited cell proliferation and caused ER stress, as well as increased the expression of ER stress markers (CHOP, 78 kDa glucose-regulated protein, activating transcription factor 6) (p < 0.05). Furthermore, RD increased the cell apoptosis rate, enhanced the expression of proapoptotic markers (B-cell lymphoma 2-associated X, Caspase 3), and decreased the expression of the antiapoptotic marker (B-cell lymphoma 2) (p < 0.05). The 4-PBA treatment and CHOP knockdown markedly alleviated RD-induced cell apoptosis. These results demonstrate that RD induces cell apoptosis by triggering ER stress and the CHOP pathway.

Background: Although methionine (Met), the first-limiting dietary amino acid, has crucial roles in growth and regulation of lipid metabolism in ducks, mechanisms underlying are not well understood. Therefore, the objective was to use dietary Met deficiency to investigate the involvement of Met in lipid metabolism and fat accumulation of Pekin ducks. Methods: A total of 150 male Pekin ducks (15-d-old, 558.5 ± 4.4 g) were allocated into 5 groups (6 replicates with 5 birds each) and fed corn and soybean meal-based diets containing 0.28%, 0.35%, 0.43%, 0.50%, and 0.58% Met, respectively, for 4 weeks. Met-deficient (Met-D, 0.28% Met) and Met-adequate (Met-A, 0.43% Met) groups were selected for subsequent molecular studies. Serum, liver, and abdominal fat samples were collected to assess the genes and proteins involved in lipid metabolism of Pekin ducks and hepatocytes were cultured in vivo for verification. Results: Dietary Met deficiency caused growth depression and excess fat deposition that were ameliorated by feeding diets with adequate Met. Serum triglyceride and non-esterified fatty acid concentrations increased (P < 0.05), whereas serum concentrations of total cholesterol, low density lipoprotein cholesterol, total protein, and albumin decreased (P < 0.05) in Met-D ducks compared to those in Met-A ducks. Based on hepatic proteomics analyses, dietary Met deficiency suppressed expression of key proteins related to fatty acid transport, fatty acid oxidation, tricarboxylic acid cycle, glycolysis/gluconeogenesis, ketogenesis, and electron transport chain; selected key proteins had similar expression patterns verified by qRT-PCR and Western blotting, which indicated these processes were likely impaired. In vitro verification with hepatocyte models confirmed albumin expression was diminished by Met deficiency. Additionally, in abdominal fat, dietary Met deficiency increased adipocyte diameter and area (P < 0.05), and down-regulated (P < 0.05) of lipolytic genes and proteins, suggesting Met deficiency may suppress lipolysis in adipocyte. Conclusion: Taken together, these data demonstrated that dietary Met deficiency in Pekin ducks resulted in stunted growth and excess fat deposition, which may be related to suppression of fatty acids transportation and hepatic catabolism.

A 2 × 6 factorial experiment was conducted to determine the influences of dietary metabolizable energy (ME) and methionine (Met) levels on growth performance, carcass traits, and plasma biochemical parameters of starter Pekin ducks from 1 to 21 days of age. A total of 600 one-day-old male Pekin ducklings were randomly assigned to 12 groups (six replicates each group and eight ducks per replicate) in a 2 × 6 two-factor arrangement. The basal Met levels of two basal diets (11.54 and 12.52 MJ/kg ME) were 0.31 and 0.29%, respectively. The crystalline L-Met was supplemented to yield six diets according to different supplemental levels (0, 0.05, 0.10, 0.15, 0.20, and 0.25%). The results showed that the body weight (BW) and average daily weight gain (ADG) were increased (p < 0.05) with increasing dietary Met levels. Dietary ME levels changed from 11.54 to 12.52 MJ/kg increased the BW and ADG (p < 0.05) as well as decreased the average daily feed intake and feed to gain ratio (p < 0.05). As the dietary Met level increased, leg muscle yield increased (p < 0.05). Conversely, increasing the dietary ME level decreased the leg muscle yield (p = 0.0024) and increased abdominal fat (p < 0.001). Meanwhile, the concentrations of total cholesterol (TCHO), high-density lipoprotein cholesterol (HDLC), and low-density lipoprotein cholesterol (LDLC) in plasma were decreased (p < 0.05) when the ME levels of diets changed from 11.54 to 12.52 MJ/kg. Meanwhile, the plasma TCHO and HDLC concentrations decreased (p < 0.05) as dietary Met levels increased. Based on the linear-broken line model, the dietary Met requirement of starter Pekin ducks from 1 to 21 days of age for optimal ADG were 0.362% (0.052% supplemental L-Met) at 11.54 MJ ME/kg and 0.468% (0.178% supplemental L-Met) at 12.52 MJ ME/kg, respectively, when crystal L-Met was supplemented to formulate the diets. This suggested that the Met requirement of starter Pekin ducks was affected by dietary ME levels. The data potentially provide theoretical support for the utilization of crystalline L-Met in duck production.

The objective of this study is to determine the effects of the dietary replacement of soybean oil (SO) with rubber seed oil (RSO) on the growth performance, carcass trait, and lipid metabolism in Pekin ducks. A total of 160 1-day-old Pekin ducks were randomly allocated to four experimental treatments and fed diets with different ratios of SO to RSO as follows: 3:0 (control), 2:1, 1:2, and 0:3. Dietary RSO supplementation had no effect on their growth performance; however, it significantly decreased the yield of abdominal fat (p < 0.05). As the dietary RSO increased, the plasma TG, CHO, LDL-C, and HDL-C contents of ducks decreased (p < 0.05). Additionally, the contents of total fat, triglycerides, and cholesterol in the liver and breast reduced in the ducks fed RSO diets (p < 0.05). Liver n-3 PUFA levels linearly increased (p < 0.05), while the n-6/n-3 PUFA ratios reduced with increasing RSO levels (p < 0.05). Moreover, dietary RSO supplementation resulted in decreased gene expressions of FABP1, ME1, SREBP1c, FASN, DGAT2, and HMGCR (p < 0.05), while there was an increased expression of the ABCA1 gene (p < 0.05) in the liver of the ducks. In conclusion, dietary RSO supplementation reduced fat deposition and enhanced n-3 PUFA levels without affecting the growth performance of Pekin ducks.