Dietary carbohydrates influence muscle texture of olive flounder Paralichthys olivaceus through impacting mitochondria function and metabolism of glycogen and protein.

The present study was conducted to estimate the effects of dietary carbohydrates on muscle quality and the underlying mechanisms. Six isonitrogenous and isolipidic diets were formulated to contain graded levels of carbohydrates (0%, 8%, 12%, 16%, 20% and 24%, respectively). These diets were named as C0, C8, C12, C16, C20 and C24, respectively. After a 10-week feeding trial, results showed that the muscle pH, liquid holding capacity (LHC) and hardness were significantly decreased by the increasing dietary carbohydrate levels. Dietary carbohydrates significantly decreased the muscle fibre diameter, and the highest value was found in the C0 group. Accumulated glycogen and degenerated mitochondrial cristae were observed in the C24 group. Significantly higher contents of protein carbonyls were observed in the C20 group and C24 group (P < 0.05). There was a significant decrease of mtDNA copy number in the C24 group compared with that in the C0 and C8 groups. The AMP/ATP ratio in muscle decreased first and then increased with the increasing dietary carbohydrate levels. The dietary incorporation of carbohydrate significantly reduced the expression of opa1, pygm and genes involved in myogenesis (myf5 and myog). Meanwhile, proteolysis-related genes (murf-1, mafbx, capn2 and ctsl), pro-inflammatory cytokines (il-6 and tnf-α) and mstn were significantly up-regulated. In the C24 group, significant increase of phosphorylation of AMPK (Thr172), up-regulation of PGC-1α and GLUT4 were observed, while the phosphorylation level of S6 (Ser235/236) was significantly decreased. It was concluded that excessive dietary carbohydrate level (24%) had negative impacts on mitochondria function and promoted glycogen accumulation, and thereafter influenced the muscle quality of olive flounder. The activation of AMPK as well as the upregulation of PGC-1α and GLUT4 was the key mechanism.

Forkhead box O1 in turbot Scophthalmus maximus: Molecular characterization, gene structure, tissue distribution and the role in glucose metabolism.

Forkhead box O1 (foxo1) is a transcription factor and plays important roles in glucose metabolism. In the present study, foxo1 in turbot Scophthalmus maximus was cloned and characterized. The siRNA of foxo1 was used to investigate the functions of foxo1 in turbot hepatocytes glucose metabolism. After that, a 10-week feeding trial with two different dietary carbohydrate levels (15% and 21%, respectively) was conducted to analyze the function of foxo1 in glucose metabolism in vivo. Results showed that the foxo1 was identified as 2176 bp (base pair) with a 2025 bp open reading frame, which encoded 675 amino acids. Sequence analysis showed that foxo1 of turbot was highly homologous to most of fishes. Tissue distribution analysis revealed that the highest expression of foxo1 was in liver. After in vitro analysis, foxo1-specific small interfering RNA (sifoxo1) treatment significantly decreased the expressions of cytosolic phosphoenolpyruvate carboxykinase (cpepck) and glucose-6-phosphatase1(g6pase1) in primary hepatocytes. Expression of mitochondrial phosphoenolpyruvate carboxykinase (mpepck) was not significantly inhibited. In contrast, the expression of glucose-6-phosphatase2 (g6pase2) increased significantly. After the in vivo study (feeding trial), with the decreased expression of foxo1 in turbot due to high dietary carbohydrate level (21%), the expression of g6pase2 was significantly upregulated. However, the expression of glucokinase (gk) was not changed significantly. These increased the level of blood glucose and hepatic glycogen. In conclusion, data from both in vitro (primary hepatocytes) and in vivo (feeding trial) showed that downregulated foxo1 in turbot could not result in significant depression of gluconeogenesis and activation of glycolysis. This could be one of the reasons why intake of high level of carbohydrate resulted in prolonged hyperglycemia in turbot.

Responses of glucosensing system to glucose in Japanese flounder Paralichthys olivaceus fed diets with different carbohydrate content.

A 10-week feeding trial was conducted to investigate the response of glucosensing system to glucose in Japanese flounder Paralichthys olivaceus (initial body weight: 7.14 ±â€¯0.10 g) fed diets with different carbohydrate content. Two experimental diets were designed as carbohydrate free (CF) and suitable carbohydrate (SC) supplementation, respectively. The dietary carbohydrate contents were 0.93% and 15.6%, respectively. After a 10-week feeding trial, a glucose tolerance test (GTT) was performed. Results showed that after the last meal in the feeding trial, the blood glucose of fish fed with diet CF peaked at 3 h (4.64 ±â€¯0.29 mM), the duration of hyperglycemia was about 5 h (1-6 h). The blood glucose in SC group peaked at 9 h (3.28 ±â€¯0.66 mM), and the duration of hyperglycemia was approximately 6 h (6-12 h). After GTT, blood glucose reached the first peak at 6 h both in the two groups, and the duration of hyperglycemia was obvious 24 h. During the 3-12 h after injection, blood glucose level in SC group was significantly higher than that in CF group. However, blood glucose level in group SC was significantly lower than that in group CF at 24 h. The blood glucose level decreased to half of the peak at 10.97 h after injection of glucose in SC group and at 27.26 h in CF group. The 6-24 h clearance ability in SC group (6.57 ±â€¯1.68%/h) was significantly higher than that in CF group (2.81 ±â€¯1.11%/h). Compared with CF diet, SC diet significantly increase the expression of glucosensing-related genes including glucose facilitative transporter type 2, glucokinase, inward rectifier K+ channel pore type 6.2, sulfonylurea receptor, carnitine palmitoyltransferase 1b, hydroxyacyl-CoA dehydrogenase, cytochrome c oxidase subunit 4, mitochondrial uncoupling protein 2a, liver X receptor, sodium/glucose co-transporter 1, a heterodimer of type 1 receptor subunits depending on T1R2 + T1R3 in liver and intestine. Meanwhile, activities of glucokinase, pyruvate kinase and glycogen synthase in liver, and hepatic glycogen content were also increased. In conclusion, glucosensing systems in Japanese flounder are responsive to dietary carbohydrate levels, especially the suitable dietary carbohydrate level, at which the glucose tolerance capacity of Japanese flounder was improved.

High level of dietary soybean oil affects the glucose and lipid metabolism in large yellow croaker Larimichthys crocea through the insulin-mediated PI3K/AKT signaling pathway.

The present study was conducted to investigate the metabolic responses of glucose and lipid in large yellow croaker Larimichthys crocea (initial weight, 36.80 ±â€¯0.39 g) to high level of dietary soybean oil. Three isonitrogenous (46% crude protein) and isolipidic (13% crude lipid) experimental diets were designed, with 100% fish oil (FO), 50% fish oil and 50% soybean oil (FS) and 100% soybean oil (SO), respectively. After a 12-week growth trial, the results showed that compared with FO group, contents n-6 PUFAs increased while the n-3 PUFAs decreased significantly both in liver and muscle in FS and SO groups. Concentrations of blood glucose, leptin, free fatty acid and total triglyceride reached the highest values in SO group, while blood insulin showed no significant difference among all groups. The gene expressions of insulin receptor substrate-2, glucose-6-phosphatase, phosphoenolpyruvate carboxykinase, fatty acid synthetase, and lipoprotein lipase increased, and the insulin receptor substrate-1, phosphotidylinsositol-3-kinase (PI3K), hexokinase, glycogen synthetase and glucose transporter 2 in liver decreased significantly in SO group. Meanwhile, the phosphorylation of protein kinase B (AKT) also decreased significantly in this group. These results suggested that high level of dietary soybean oil depressed PI3K/AKT signaling pathway, and then affected glucose and lipid metabolism by glycolysis, gluconeogenesis, glucose transportation, glycogenesis and lipogenesis.

Chronic stress of high dietary carbohydrate level causes inflammation and influences glucose transport through SOCS3 in Japanese flounder Paralichthys olivaceus.

Carnivorous fish is thought to be high-glucose intolerance. But the reasons were still unclear. The aim of the present study is to investigate the effects of high level of dietary carbohydrate on the survival, growth and immune responses of Paralichthys olivaceus, and the underlying molecular mechanism related to the immune and glucose metabolism. P. olivaceus were fed with 8%, 16% and 24% of dietary carbohydrate for 10 weeks, respectively. After that, a glucose tolerance test (GTT) was conducted. Results showed that excessive (24%) dietary carbohydrate significantly decreased the growth and glucose tolerance ability according to the GTT. It significantly increased hepatic NADPH oxidase activity and malondialdehyde content and serum contents of IL-6 and advanced glycation end products. The expressions of glucose transport-relevant genes in liver and the content of related hormones in serum were analyzed. In conclusion, it was confirmed that IL-6 increased the expression of suppressor of cytokine signaling 3 (SOCS3) and regulated the downstream targets of PI3K-AKT mediated signal transduction, and then downregulated the glucose transporter 2 activity in liver of P. olivaceus fed diet with excessive carbohydrate level. It was suggested that SOCS3 served as a bridge between immune response and glucose metabolism in P. olivaceus.

Comparatively study on the insulin-regulated glucose homeostasis through brain-gut peptides in Japanese flounder Paralichthys olivaceus after intraperitoneal and oral administration of glucose.

The present study comparatively analyzed the blood glucose and insulin concentration, the temporal and spatial expression of brain-gut peptides and the key enzymes of glycolysis and gluconeogenesis in Japanese flounder by intraperitoneal injection (IP) and oral administration (OR) of glucose. Samples were collected at 0, 1, 3, 5, 7, 9, 12, 24 and 48 h after IP and OR glucose, respectively. Results showed that the hyperglycemia lasted for about 10 h and 21 h in OR and IP group, respectively. The serum insulin concentration significantly decreased at 3 h (1.58 ±â€¯0.21 mIU/L) after IP glucose. However, it significantly increased at 3 h (3.37 ±â€¯0.341 mIU/L) after OR glucose. The gene expressions of prosomatostatin, neuropeptide Y, cholecystokinin precursor and orexin precursor in the brain showed different profiles between the OR and IP group. The OR not IP administration of glucose had significant effects on the gene expressions of preprovasoactive intestinal peptide, pituitary adenylate cyclase activating polypeptide and gastrin in intestine. In conclusion, brain-gut peptides were confirmed in the present study. And the serum insulin and the brain-gut peptides have different responses between the IP and OR administration of glucose. The OR could stimulate the brain-gut peptide expressions, which have effects on the insulin secretion and then regulate the blood glucose levels. However, in IP group, there is little chance to stimulate brain-gut peptide expression to influence the insulin secretion, which leads to a longer hyperglycemia.

Comparative microarray profile of the hepatopancreas in the response of "Huanghai No. 2" Fenneropenaeus chinensis to white spot syndrome virus.

White spot syndrome virus (WSSV) infects all shrimp species and is the greatest detriment to shrimp culture. To better understand the mechanism of molecular responses to WSSV infection in "Huanghai No. 2" Fenneropenaeus chinensis, a microarray technique was used. Microarray gene expression profiling of 59,137 unigenes identified Differentially Expressed Genes (DEGs) both in live and moribund shrimp at early, peak and late phases. In live shrimp, 1307, 1479 and 1539 DEGs were obtained in the early, peak and late phase, respectively. Meanwhile, 1536, 2181 and 1591 DEGs were obtained in moribund shrimp. Twenty known annotation genes are uniquely expressed in the late phase of live shrimp, including adhesion regulating molecule 1, arginine kinase, BUD31 homolog, and QM. Compared to WSSV-susceptible shrimp, 75 known annotation genes are uniquely expressed in WSSV-resistant shrimp, including arginine kinase, BUD31 homolog, clottable protein 2, caspase 2, cathepsin C, calnexin, HMGBb, Histone 3, and selenoprotein M. The gene expression patterns of the infected shrimp were altered by WSSV infection. To further confirm the expression of differentially expressed genes, real-time RT-PCR was performed to test six randomly selected genes. The data will provide valuable information to understand the immune mechanism of shrimp's response to WSSV.