Phosphoenolpyruvate Carboxykinase mRNA Levels Research Articles

Urtica dioica (Gazaneh) distillate restores glucose metabolism in diabetic rats.

Diabetes has become an important health problem in the world. Natural agents, with antidiabetic property, are potential candidates for improving diabetes. Urtica Dioica Distillate (UDD) or Araghe Gazaneh is widely used for the treatment of diabetes as per traditional medicine. Despite the tremendous use of UDD as an antidiabetic compound in folk medicine, the antidiabetic effects of UDD has been neglected by medical scientists. In this study, we aimed to evaluate the effects of UDD on the glucose metabolism in diabetic rats. A total of 24 male rats were divided equally into four groups, two treatment and two control groups, each containing normal or Streptozotocin (STZ)-induced diabetic rats. During 4 weeks, control and treatment rats received water or UDD, respectively. Fasting blood sugar (FBS), HbA1c, serum creatinine, blood urea nitrogen, and specific activities of hepatic enzymes including glucokinase (GK), hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), and muscle glucose transporter type 4 (GLUT4) and liver phosphoenolpyruvate carboxykinase (PEPCK) mRNA levels were measured. FBS and HbA1c increased in diabetic groups. Treatment with UDD significantly lowered FBS and prevented weight loss. Decreased FBS level was associated with higher activity levels of GK and HK in UDD-treated diabetic rats. G6PD-specific activity decreased in diabetic control rats compared to nondiabetic ones, but UDD treatment improved it to the normal levels. A significant decrease in the expression level of GLUT4 was observed in diabetic control rats compared to nondiabetic ones, but UDD increased it to the normal levels. These findings suggest that UDD might exert therapeutic effects against diabetes by improving glucose metabolism and can be used as an alternative or complementary medicine for the treatment of diabetic patients.

DNA methylation of pck1 might contribute to the programming effects of early high-carbohydrate diets feeding to the glucose metabolism across two generations in zebrafish (Danio rerio).

The purpose of this study is to assess the effects of early high-carbohydrate stimulus on glucose metabolism in zebrafish (Danio rerio) over two generations and explore the mechanisms that explain those nutritional programming effects via epigenetic modifications. The larvae were delivered a high-carbohydrate diet (53.66%) that was used as an early nutritional stimulus from the first feeding to the end of the yolk sac (FF) and 5days after yolk-sac exhaustion (YE). The larvae (F0) and their offspring (F1) were then both fed the control diet (22.69%) until adulthood (15weeks), and they were challenged with a high-carbohydrate diet (35.36%) at the 16th week. The results indicated that early stimulus immediately raised the mRNA levels of genes involved in glycolysis and gluconeogenesis. At the end of F0 challenge, both treatment groups decreased the plasma glucose levels, increased the expression levels of glucokinase (gck), and inhibited the mRNA during gluconeogenesis. When challenged in F1, the glucose levels were lower in FF (F1), and the mRNA levels of phosphoenolpyruvate carboxykinase 1 (pck1) were decreased in FF (F1) and YE (F1). Besides, in both experimental groups (F0 and F1), the CpG island of pck1 maintained lower levels of hypermethylated expression from F0 adult, 24h post-fertilization embryo, to F1 adult. In conclusion, these results indicated that an early high-carbohydrate stimulus could significantly reprogram glucose metabolism in adult zebrafish, that those modifications could be partially transmitted to the next generation, and that the DNA methylation of pck1 might work as a stable epigenetic marker to contribute to those processes.

Different responses to glucose overload between two strains of largemouth bass (Micropterus salmoides).

In order to improve the glucose utilization capacity of largemouth bass (Micropterus salmoides), responses to glucose overload between two strains (Y: breeding strain; W: wild strain) were compared at 0, 6, 12, and 24 h after glucose injection (1.67 g/kg). The data revealed that plasma glucose in the Y strain (<12 h) recovered faster than in the W strain (12 h), with the Y strain secreted more insulin within 6 h post-injection. Triglyceride (TG) and low-density lipoprotein-cholesterol (VLDL-CH) content in the Y strain increased, peaking at 12 h, then decreased, whereas the W strain’s TG content was not affected and VLDL-CH content decreased. The hepatic and muscular fatty acid synthetase, liver x receptor-1, and sterol regulatory element-binding protein expressions were consistent with the TG content change. Both strains’ liver and muscle glycogen contents exhibited similar trends to that of the glycogen synthase gene—increasing, then declining, and peaking at 6 and 12 h. The expression levels of hepatic and muscular phosphofructokinase and pyruvate kinase in the Y strain increased, peaking at 12 h. In the W strain, they were suppressed and reached the minimum at 24 h. The mRNA levels of hepatic and muscular phosphoenolpyruvate carboxykinase and glucose-6-phosphatase were enhanced and peaked at 24 h in both strains, hepatic isocitrate dehydrogenase-1, and α-ketoglutarate dehydrogenase complex expression increased after declining, peaking at 12 and 24 h. Two genes in the W strain’s muscles showed a similar trend. Both strains’ transcriptome results identified seven common functional genes for resistance to hyperglycemia that were involved in the circadian rhythm pathway, which is a suggested key pathway for coping with hyperglycemia. Furthermore, 48 differential genes were identified between the two strains, and these genes were enriched in the TGF-beta and cell cycle signaling pathways, indicating that these pathways may be key factors affecting the differential responses to glucose overload. We conducted a comprehensive comparison of glucose overload molecular responses between two strains of M. salmoides, and the results can provide a promising strategy to improve the glucose utilization capacity of M. salmoides based on advantageous pre-existing traits.

Organic chromium derived from the chelation of Ganoderma lucidum polysaccharide and chromium (III) alleviates metabolic syndromes and intestinal microbiota dysbiosis induced by high-fat and high-fructose diet

Organic chromium is of great interest and has become an important chromium supplement resource in recent years because of its low toxicity and easy absorption. In our previous study, we synthesized a novel organic chromium [GLP-Cr] through the chelation of Ganoderma lucidum polysaccharide and chromium (III). The purpose of this study was to investigate the beneficial effects of GLP-Cr on the improvement of metabolic syndromes (MetS) in mice fed with a high-fat and high-fructose diet (HFHFD) and its mechanism of action. The results indicated that oral administration of GLP-Cr inhibited the excessive exaltation of body weight, glucose tolerance, fasting blood glucose and lipid levels, hepatic total cholesterol (TC), triglyceride (TG) levels caused by HFHFD. Besides, 16S rRNA amplicon sequencing showed that GLP-Cr intervention evidently ameliorated intestinal microbiota dysbiosis by changing the proportions of some intestinal microbial phylotypes. In addition, correlation network-based analysis indicated that the key intestinal microbial phylotypes were closely related to biochemical parameters associated with MetS under GLP-Cr intervention. Liver metabolomics analysis suggested that GLP-Cr intervention significantly regulated the levels of some biomarkers involved in alpha-linolenic acid metabolism, fatty acid biosynthesis, steroid hormone biosynthesis, glycerophospholipid metabolism, glycerolipid metabolism, steroid hormone biosynthesis, primary bile acid biosynthesis, and so on. Moreover, GLP-Cr intervention regulated liver mRNA levels of key genes associated with glucose and lipid metabolism. The mRNA level of glucose transporter type 4 (Glut4) was markedly increased by GLP-Cr intervention, and the mRNA levels of phosphoenolpyruvate carboxykinase (Pepck) and glucose-6-phosphatase (G6Pase) in the liver were significantly decreased. Meanwhile, GLP-Cr intervention significantly decreased hepatic mRNA levels of cluster of differentiation 36 (Cd36), acetyl-CoA carboxylase 1 (Acc1) and sterol regulatory element binding protein-1c (Srebp-1c), indicating that GLP-Cr intervention inhibited the excessive accumulation of free fatty acids in the liver. These findings suggest that the prevention of hyperglycemia and dyslipidemia by GLP-Cr may be closely related to the regulation of gut microbial composition and hepatic metabolic pathways, thus GLP-Cr can be serving as a functional component in the prevention of MetS.

Reduced insulin signaling and high glucagon in early insulin resistance impaired fast-fed regulation of renal gluconeogenesis via insulin receptor substrate.

Gluconeogenesis is one of the key processes through which the kidney contributes to glucose homeostasis. Urinary exosomes (uE) have been used to study renal gene regulation noninvasively in humans and rodents. Recently, we demonstrated fast-fed regulation of phosphoenolpyruvate carboxykinase (PEPCK), the rate-limiting enzyme for gluconeogenesis, in human uE. The regulation was impaired in subjects with early insulin resistance. Here, we studied primary human proximal tubule cells (hPT) and human uE to elucidate a potential link between insulin resistance and fast-fed regulation of renal PEPCK. We demonstrate that fasted hPTs had higher PEPCK and insulin receptor substrate-2 (IRS2) mRNA and protein levels, relative to fed cells. The fast-fed regulation was, however, attenuated in insulin receptor knockdown (IRKO) hPTs. The IRKO was confirmed by the blunted insulin-induced response on PEPCK, PGC1α, p-IR, and p-AKT expression in IRKO cells. Exosomes secreted by the wild-type or IRKO hPT showed similar regulation to the respective hPT. Similarly, in human uE, the relative abundance of IRS-2 mRNA (to IRS1) was higher in the fasted state relative to the fed condition. However, the fast-fed difference was absent in subjects with early insulin resistance. These subjects had higher circulating glucagon levels relative to subjects with optimal insulin sensitivity. Furthermore, in hPT cells, glucagon significantly induced PEPCK and IRS2 gene, and gluconeogenesis. IR knockdown in hPT cells further increased the gene expression levels. Together the data suggest that reduced insulin sensitivity and high glucagon in early insulin resistance may impair renal gluconeogenesis via IRS2 regulation.

Renal gluconeogenesis induction by glucagon and insulin receptor deletion in humans

Gluconeogenesis is one of the key processes through which the kidney contributes to glucose homeostasis. Recently, we provided evidence to suggest that early insulin resistance may impair fast‐fed regulation of Phosphoenolpyruvate Carboxykinase (PEPCK), rate‐limiting enzyme for gluconeogenesis in humans. To confirm this and to elucidate a link between insulin resistance and fast‐fed regulation of PEPCK in the kidneys, we studied primary proximal tubule cells (hPT) and urinary extracellular vesicles (uEVs) from humans. We demonstrate that fasted hPTs had higher PEPCK and insulin receptor substrate‐2 (IRS2) mRNA levels than fed cells. The fast‐fed regulation was, however, attenuated in insulin receptor knockdown hPTs (IRKO). The insulin receptor knockdown was confirmed by a blunted insulin‐induced suppression in PEPCK and PGC1α mRNA and p‐IR and p‐AKT protein expression in IRKO cells. Exosomes secreted by the wild‐type or IRKO hPT showed similar regulation as observed in the respective hPT. Similarly, in human uE, the relative abundance of IRS‐2 mRNA (to IRS1) was higher in the fasted state relative to the fed condition. However, the fast‐fed difference was absent in subjects with early insulin resistance. These subjects had higher circulating glucagon levels relative to subjects with optimal insulin sensitivity. Furthermore, in hPT cells, glucagon significantly induced PEPCK mRNA, IRS2 mRNA, and gluconeogenesis. Insulin receptor deletion further increased their levels. Together the data suggest that reduced insulin sensitivity and high glucagon in early insulin resistance may impair renal gluconeogenesis via IRS2 regulation. Funding from ICMR‐CARE and SGPGI intramural to ST and CSIR to RS.

Nutrient metabolism in the liver and muscle of juvenile blunt snout bream (Megalobrama amblycephala) in response to dietary methionine levels

A 75-day rearing trial was designed to study the response of juvenile Megalobrama amblycephala to dietary methionine (Met) levels. Three practical diets with graded Met levels (0.40%, 0.84% and 1.28% dry matter) were prepared to feed the juvenile fish. The results showed that the 0.84% Met diet significantly improved the growth compared with 0.40% diets. Compared with 0.84% and 1.28% Met, 0.40% Met significantly increased the hepatic lipid content, while decreasing the muscular lipid and glycogen contents. 0.40% Met decreased the protein levels of phospho-Eukaryotic initiation factor 4E binding protein-1 (p-4e-bp1), 4e-bp1 and Ribosomal protein S6 kinase 1 in the liver, compared with 0.84% diet, while an increasing trend was observed in the muscle. Met supplementation tended to decrease and increase lipid synthesis in the liver and muscle, respectively, via changing mRNA levels of sterol regulatory element-binding protein 1, fatty acid synthetase and acetyl-CoA carboxylase. 1.28% dietary Met promoted fatty acid β-oxidation and lipolysis in both the liver and muscle by increasing carnitine palmitoyl transferase 1, peroxisome proliferator activated receptor alpha, lipoprotein lipase and lipase mRNA levels. Compared with 0.40% and 0.84% dietary Met, 1.28% Met enhanced the mRNA levels of hepatic gluconeogenesis related genes phosphoenolpyruvate carboxykinase (pepck), and glucose-6-phosphatase, and muscular glycolysis related genes phosphofructokinase (pfk), and pyruvate kinase (pk). The mRNA levels of hepatic pfk, pk and glucokinase were markedly downregulated by 1.28% Met compared with 0.84% level. Muscular pepck, glycogen synthase, and hepatic glucose transporters 2 mRNA levels were induced by 1.28% Met. Generally, deficient Met level decreased the growth of juvenile Megalobrama amblycephala, and the different nutrient metabolism responses to dietary Met were revealed in the liver and muscle.

Interactions of dietary carbohydrate and vitamin D3 on growth performance, insulin signaling pathway and glucose metabolism in juvenile abalone Haliotis discus hannai

A 115-day feeding trial was conducted to investigate the effect of dietary carbohydrate and vitamin D3 levels, and their interactions on growth performance, body composition, insulin signaling pathway and glucose metabolism in juvenile abalone Haliotis discus hannai (initial body weight: 2.06 ± 0.01 g). Nine experimental diets in a 3 × 3 two-factor design were formulated with different carbohydrate levels (27.5%, 32% and 36.5%) and vitamin D3 levels (0, 1000 IU/kg and 5000 IU/kg). Results showed that the highest specific growth rate (SGR) of abalone was observed in the treatment with 36.5% of dietary carbohydrate and 1000 IU/kg of vitamin D3. The serum glucose, body crude lipid and muscle glycogen contents of abalone significantly increased with increasing dietary carbohydrate levels (P < 0.05). Diet with 1000 IU/kg of vitamin D3 significantly reduced the serum glucose and body crude lipid content (P < 0.05). Significant interactions between dietary carbohydrate and vitamin D3 were observed in SGR and crude lipid content (P < 0.05). Diet with 36.5% carbohydrate up-regulated the expression of pyruvate kinase, glucose-6-phosphate dehydrogenase (G6PDH), insulin receptor (IR), insulin receptor substrate (IRS), protein kinase B (Akt), glycogen synthase and glycogen phosphorylase (PYG), whereas decreased the phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6P) and glycogen synthesis kinase-3β. Diet with 1000 IU/kg of vitamin D3 significantly up-regulated the hexokinase, IR, IRS, Akt and glucose transporter 1 mRNA levels, whereas decreased the G6PDH (P < 0.05). The interaction between dietary carbohydrate and vitamin D3 were significant on PEPCK, G6P, G6PDH and PYG mRNA levels (P < 0.05). In conclusion, dietary carbohydrates and vitamin D3 could affect glucose metabolism by regulating glycolysis, gluconeogenesis and pentose phosphate pathway in abalone. Besides, they triggered the insulin signaling pathway, and further influence glucose transport and glycogen synthesis, which could promote the utilization of carbohydrate in abalone.

Melatonin stimulates transcription of the rat phosphoenolpyruvate carboxykinase gene in hepatic cells.

Melatonin plays physiological roles in various critical processes, including circadian rhythms, oxidative stress defenses, anti-inflammation responses, and immunity; however, the current understanding of the role of melatonin in hepatic glucose metabolism is limited. In this study, we examined whether melatonin affects gene expression of the key gluconeogenic enzyme, phosphoenolpyruvate carboxykinase (PEPCK). We found that melatonin treatment increased PEPCK mRNA levels in rat highly differentiated hepatoma (H4IIE) cells and primary cultured hepatocytes. In addition, we found that melatonin induction was synergistically enhanced by dexamethasone, whereas it was dominantly inhibited by insulin. We also report that the effect of melatonin was blocked by inhibitors of mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK), RNA polymerase II, and protein synthesis. Furthermore, the phosphorylated (active) forms of ERK1 and ERK2 (ERK1/2) increased 15min after melatonin treatment. We performed luciferase reporter assays to show that melatonin specifically stimulated promoter activity of the PEPCK gene. Additional reporter analysis using 5'-deleted constructs revealed that the regulatory regions responsive to melatonin mapped to two nucleotide regions, one between -467 and -398 nucleotides and the other between -128 and +69 nucleotides, of the rat PEPCK gene. Thus, we conclude that melatonin induces PEPCK gene expression via the ERK1/2 pathway at the transcriptional level, and that induction requires denovo protein synthesis.

Culture salinity alters dietary protein requirement, whole body composition and nutrients metabolism related genes expression in juvenile Genetically Improved Farmed Tilapia (GIFT) (Oreochromis niloticus)

An 8-week experimental trial was designed to explore the effects of dietary protein level (DPL) and salinity on the growth performance and physiological parameters of genetically improved farmed tilapia (GIFT) juveniles (Oreochromis niloticus). Six diets were formulated with different dietary protein levels (18.20%, 24.32%, 30.60%, 36.42%, 43.27% and 49.49%) and two salinity levels (0and 8‰) in a 6 × 2 factorial design. 720GIFT juveniles (initial weight 3.25 g) were distributed into 36 plastic tanks (180 L) and fed each of the diets four times daily to satiation. The results suggested that the growth performance was increased significantly with increasing DPLs up to a certain level (36.42%) in freshwater and brackish water. The whole-body lipid content of the fish decreased significantly with increasing salinity, while the protein content increased (P < 0.05). However, the protein content increased significantly and the lipid, moisture and ash contents were not significantly affected by the increase in DPL. With the increasing DPL at two salinities, the expression levels of hepatic insulin-like growth factor-1 increased significantly, reaching a maximum at 36.42% DPL. With the increasing DPL at the two salinity levels, the expression level of oligopeptide transporter 2 mRNA significantly decreased (P < 0.05). The target of rapamycin, 4E binding protein 1, ribosomal protein S6 and oligopeptide transporter 1β mRNA levels were not significantly affected by the DPL, but they increased significantly with an increase in salinity (P < 0.05). Regarding glucolipid metabolism, glycogen synthase, glucokinase and glucose transporter 2 mRNA levels decreased with an increase in DPL (P < 0.05). The fatty acid synthase and glucokinase mRNA levels decreased with an increase in salinity, while glycogen synthase and glucose transporter 2 mRNA levels increased (P < 0.05). The phosphoenolpyruvate carboxykinase mRNA level was not significantly affected by the DPL, but it increased with increasing salinity (P < 0.05). For the feed conversion ratio and weight gain rate, the broken-line model analysis showed that the optimal protein requirements at 8‰ salinity were higher than those at 0‰ salinity and the protein requirement were 37.70%, 37.86% (8‰) and 33.70%, 38.21% (0‰), respectively. GIFT juveniles exhibited improved glycogen synthesis at 8‰ salinity and fat deposition at 0‰ salinity.

Berberine reduces gut-vascular barrier permeability via modulation of ApoM/S1P pathway in a model of polymicrobial sepsis

AimsThe hyperpermeability of gut-vascular barrier (GVB) plays a role in gut-derived sepsis. The goal of this study was to evaluate if berberine might improve hepatic apolipoprotein M (ApoM) generation and raise plasma ApoM level to protect the compromised GVB. Materials and methodsThe compromised GVB was induced by sepsis. Hepatic ApoM mRNA and phosphoenolpyruvate carboxykinase (PEPCK) mRNA and plasma ApoM level were assayed by qRT-PCR and ELISA, respectively. The permeability of intestinal capillary in vivo and of rat intestinal microvascular endothelial cells (RIMECs) in vitro was assayed by FITC-dextran. The blood glucose was detected by a glucometer. Plasma insulin, TNF-α and IL-1β were assayed by ELISA. The plasmalemma vesicle-associated protein-1 (PV1), β-catenin and occludin in RIMECs were assayed by Western blot. Key findingsSepsis decreased hepatic ApoM mRNA and plasma ApoM level, but raised hepatic PEPCK mRNA and plasma glucose, insulin, TNF-α, and IL-1β levels. The increased vascular endothelial permeability was abrogated by recombinant rat ApoM in vivo or ApoM-bound S1P in vitro. ApoM-bound S1P decreased PV1 but increased occludin and β-catenin expression in LPS-treated RIMECs. Berberine in a dose-dependent manner raised hepatic ApoM mRNA and plasma ApoM level, but decreased septic hyperglycemia, insulin resistance and plasma TNF-α and IL-1β levels. Berberine reduced sepsis-induced PEPCK and TLR4 mRNA overexpression in the liver. SignificanceThis study demonstrated berberine inhibited TLR4-mediated hyperglycemia, insulin resistance and proinflammatory molecule production, thereby increasing ApoM gene expression and plasma ApoM. Berberine protected the damaged GVB via modulation of ApoM/S1P pathway.

Circular RNA HIPK3 contributes to hyperglycemia and insulin homeostasis by sponging miR-192-5p and upregulating transcription factor forkhead box O1.

It has been shown that circular RNAs, a class of non-coding RNA molecules, play an important role in the regulation of glucose and lipid homeostasis. In the present study, we sought to investigate the function of circular RNA HIPK3 (circHIPK3) in diabetes-associated metabolic disorders, including hyperglycemia and insulin resistance. Results show that oleate stimulated circHIPK3 increase, and that circHIPK3 enhanced the stimulatory effect of oleate on adipose deposition, triglyceride (TG) content, and cellular glucose content in HepG2 cells. MiR-192-5p was the potential target of circHIPK3, since circHIPK3 significantly decreased miR-192-5p mRNA level, whereas anti-circHIPK3 significantly increased miR-192-5p mRNA level. Further study shows that transcription factor forkhead box O1 (FOXO1) was a downstream regulator of miR-192-5p, since miR-192-5p significantly decreased FOXO1 expression, whereas circHIPK3 significantly increased FOXO1 expression. Notably, the inhibitory effect of miR-192-5p was significantly reversed by circHIPK3. In vivo study shows that anti-miR-192-5p significantly increased blood glucose content, which was significantly inhibited by FOXO1 shRNA. MiR-192-5p significantly decreased adipose deposition and TG content in HepG2 cells, which was significantly reversed by the co-treatment with circHIPK3. Forskolin/dexamethasone (FSK/DEX) significantly increased cellular glucose, mRNA level of phosphoenolpyruvate carboxykinase (PEPCK) and glucose 6-phosphatase (G6Pase), and this stimulatory effect of FSK/DEX was significantly inhibited by miR-192-5p. In the presence of circHIPK3, however, the inhibitory effect of miR-192-5p was totally lost. In summary, the present study demonstrated that circHIPK3 contributes to hyperglycemia and insulin resistance by sponging miR-192-5p and up-regulating FOXO1.

FXR activation alleviates tacrolimus-induced post-transplant diabetes mellitus by regulating renal gluconeogenesis and glucose uptake

BackgroundTacrolimus (FK506)-induced diabetes mellitus is one of the most important factors of post-transplant diabetes mellitus (PTDM). However, the detailed mechanisms underlying PTDM are still unclear. Farnesoid X receptor (FXR) regulates glycolipid metabolism. The objective of this study was to explore whether FXR is involved in the development of tacrolimus-induced diabetes mellitus.MethodsAfter C57BL/6J mice were treated with tacrolimus (FK506) for 3 months, the fasting blood glucose levels, body weights, renal morphological alterations, and mRNA expression levels of phosphoenolpyruvate carboxykinase (PEPCK) and glucose transporter 2 (GLUT2) among the control group, the FK506 group and the FK506 + GW4064 (a FXR agonist) group (n = 7) were measured. The intracellular location of peroxisome proliferator activated receptor γ coactivator-1α (PGC1α) and forkhead box O1 (FOXO1) was detected by immunofluorescence. Human renal cortex proximal tubule epithelial cells (HK-2) were treated with 15 μM FK506 or 4 μM FXR agonist (GW4064) for 24, 48 and 72 h, and the expression levels of FXR, gluconeogenesis and glucose uptake, representing the enzymes PEPCK and GLUT2, were detected with real-time PCR and western blot analyses. Finally, the mRNA levels of PEPCK and GLUT2 in HK-2 cells were measured after FXR was upregulated.ResultsFK506 significantly inhibited the mRNA and protein levels of FXR at 48 h and 72 h in HK-2 cells (P < 0.05). Meanwhile, FK506 promoted gluconeogenesis and inhibited glucose uptake in HK-2 cells (P < 0.05). However, overexpression of FXR in transfected HK-2 cell lines significantly inhibited gluconeogenesis and promoted glucose uptake (P < 0.05). The FXR agonist GW4064 significantly decreased the fasting blood glucose in mice challenged with FK506 for 3 months (P < 0.05), inhibited gluconeogenesis (P < 0.05) and significantly promoted glucose uptake (P < 0.05). Immunofluorescence staining and western blot analyses further revealed that FXR activation may affect the translocation of PGC1α and FOXO1 from the nucleus to the cytoplasm.ConclusionsFXR activation may mitigate tacrolimus-induced diabetes mellitus by regulating gluconeogenesis as well as glucose uptake of renal cortex proximal tubule epithelial cells in a PGC1α/FOXO1-dependent manner, which may be a potential therapeutic strategy for the prevention and treatment of PTDM.

Microarray analysis reveals the inhibition of intestinal expression of nutrient transporters in piglets infected with porcine epidemic diarrhea virus

Porcine epidemic diarrhea virus (PEDV) infection can induce intestinal dysfunction, resulting in severe diarrhea and even death, but the mode of action underlying these viral effects remains unclear. This study determined the effects of PEDV infection on intestinal absorption and the expression of genes for nutrient transporters via biochemical tests and microarray analysis. Sixteen 7-day-old healthy piglets fed a milk replacer were randomly allocated to one of two groups. After 5-day adaption, piglets (n = 8/group) were orally administrated with either sterile saline or PEDV (the strain from Yunnan province) at 104.5 TCID50 (50% tissue culture infectious dose) per pig. All pigs were orally infused D-xylose (0.1 g/kg BW) on day 5 post PEDV or saline administration. One hour later, jugular vein blood samples as well as intestinal samples were collected for further analysis. In comparison with the control group, PEDV infection increased diarrhea incidence, blood diamine oxidase activity, and iFABP level, while reducing growth and plasma D-xylose concentration in piglets. Moreover, PEDV infection altered plasma and jejunal amino acid profiles, and decreased the expression of aquaporins and amino acid transporters (L-type amino acid transporter 1, sodium-independent amino acid transporter, B(°,+)-type amino acid transport protein, sodium-dependent neutral amino acid transporter 1, sodium-dependent glutamate/aspartate transporter 3, and peptide transporter (1), lipid transport and metabolism-related genes (lipoprotein lipase, apolipoprotein A1, apolipoprotein A4, apolipoprotein C2, solute carrier family 27 member 2, solute carrier family 27 member 4, fatty acid synthase, and long-chain acyl-CoA synthetase (3), and glucose transport genes (glucose transporter-2 and insulin receptor) in the jejunum. However, PEDV administration increased mRNA levels for phosphoenolpyruvate carboxykinase 1, argininosuccinate synthase 1, sodium/glucose co-transporter-1, and cystic fibrosis transmembrane conductance regulator in the jejunum. Collectively, these comprehensive results indicate that PEDV infection induces intestinal injury and inhibits the expression of genes encoding for nutrient transporters.

Dietary methionine affects growth and the expression of key genes involved in hepatic lipogenesis and glucose metabolism in cobia (Rachycentron canadum)

Juvenile cobia (Rachycentron canadum), a carnivorous fish, was chosen to examine the effects of methionine (Met) supplementation of low fishmeal diets on growth and the expression of key genes related to hepatic lipogenesis and glucose metabolism. Seven isonitrogenous and isolipidic diets were formulated to contain 7.2, 9.0, 10.0, 12.4, 14.1, 16.3 and 18.6 g/kg dietary Met levels. Healthy cobia (initial average weight ± SE = 9.79 ± 0.04 g) were randomly assigned into seven groups in triplicate with 40 individuals per sea cage for 8 weeks. The results showed that the weight gain rate (WGR) of fish increased as the dietary Met was increased up to 12.4 g/kg and then decreased. The expression of insulin‐like growth factor‐I (IGF‐I) and target of rapamycin (TOR) genes was inhibited by the Met‐deficient diets. The mRNA levels of fatty acid synthase (FAS) and sterol regulatory element‐binding protein‐1 (SREBP1) genes increased with increasing dietary Met until 12.4 g/kg after which the former remained constant, whereas the latter decreased from 12.4 to 18.6 g/kg Met. The mRNA levels of pyruvate kinase (PK) increased as the dietary Met levels increased, reaching a peak at 12.4 g/kg, and then decreased with further increases in Met. However, the mRNA levels of phosphoenolpyruvate carboxykinase (PEPCK) decreased as the dietary Met was increased up to 12.4 g/kg and then remained constant from 12.4 to 18.6 g/kg Met. In conclusion, based on the straight broken‐line analysis of WGR against dietary Met levels, the optimal dietary Met requirement for juvenile cobia was estimated to be 12.4 g/kg (26.9 g/kg dietary protein). A diet with the optimal level of Met induces fish growth and is associated with increased IGF‐I and TOR.

Starch to protein ratios in practical diets for genetically improved farmed Nile tilapia Oreochromis niloticus: Effects on growth, body composition, peripheral glucose metabolism and glucose tolerance

The present study was performed to evaluate the effects of dietary starch to protein ratios on growth, feed utilization, body composition, peripheral glucose metabolism, and glucose tolerance of genetically improved farmed Nile tilapia (GIFT) Oreochromis niloticus. Three iso-lipidic (5.54%) practical diets were formulated with increased starch levels at the cost of protein provision, being designated as diets LSP (13.1% starch and 38.2% protein), MSP (22.5% starch and 34.5% protein), and HSP (32.3% starch and 30.5% protein). Two hundred and forty tilapia juveniles (initial mean body weight: 23 g/fish) were maintained in 250 L rectangular tanks at 20 fish per tank and were fed experimental diets to apparent satiation for 8 weeks. Approximately 24 h after the last feeding, total fish weight of each tank was measured, and the fish number was counted. Thirty-six fish per treatment (9 per tank) were sampled to analyze the body composition, blood biochemistry, biometric indices, glucose metabolic enzymatic activities, and gene expression of the liver and white muscle. Remaining fish (36 per treatment) were subjected to a glucose tolerance test. The results showed that the mean body weight of the LSP fish (145 g/fish) was higher than that of the other treatments (130–133 g/fish). Protein efficiency ratio and protein production value were improved in tilapia that were fed diets with increased starch level from 13.1% to 32.3%. The lipid percentages and glycogen concentrations in the liver and white muscle were elevated when dietary starch level was increased from 13.1% to 32.3%. Compared with the LSP fish, the expression of representative genes involved in glucose transport (glucose transporter 2, glut2), glycolysis (muscle type a of phosphofructokinase, pfkma), glycogenesis (glycogen synthase 2, gys2), and lipogenesis (acetyl-coenzyme A carboxylase α, accα; fatty acyl synthase, fas) were upregulated in the liver of the MSP fish. The mRNA levels of hepatic gluconeogenic phosphoenolpyruvate carboxykinase 2 (pck2) and glucose-6-phosphatase a2 were 0.68- and 0.37-fold lower in the HSP fish than in the LSP fish. In the white muscle, the mRNA levels of glut4, pfkma, pfkmb, gys1 and accβ were 1.59-, 1.66-, 1.55-, 1.38- and 2.01-fold higher in the HSP fish than in the LSP fish. Activity of PFK in the liver and the muscle along with hepatic PCK activity followed similar trends to the expression of their corresponding encoding genes. Though the MSP and HSP fish obtained higher basal plasma glucose levels than the LSP fish, they took less time to restore plasma glucose after a glucose load. Thus, it was concluded that tilapia metabolically adapted well to an increase in the dietary starch level from 13.1% to 32.3% by promoting peripheral glucose storage and utilization, subsequently enhancing glucose tolerance and improving protein retention, although elevated levels of starch reduced their growth.

Oral Phenelzine Treatment Mitigates Metabolic Disturbances in Mice Fed a High-Fat Diet.

Novel mechanisms and health benefits have been recently suggested for the antidepressant drug phenelzine (PHE), known as a nonselective monoamine oxidase inhibitor. They include an antilipogenic action that could have an impact on excessive fat accumulation and obesity-related metabolic alterations. We evaluated the metabolic effects of an oral PHE treatment on mice fed a high-fat diet (HFD). Eleven-week-old male C57BL/6 mice were fed a HFD and either a 0.028% PHE solution (HFD + PHE) or water to drink for 11 weeks. PHE attenuated the increase in body weight and adiposity without affecting food consumption. Energy efficiency was lower in HFD + PHE mice. Lipid content was reduced in subcutaneous fat pads, liver, and skeletal muscle. In white adipose tissue (WAT), PHE reduced sterol regulatory element-binding protein-1c and phosphoenolpyruvate carboxykinase mRNA levels, inhibited amine-induced lipogenesis, and did not increase lipolysis. Moreover, HFD + PHE mice presented diminished levels of hydrogen peroxide release in subcutaneous WAT and reduced expression of leukocyte transmigration markers and proinflammatory cytokines in visceral WAT and liver. PHE reduced the circulating levels of glycerol, triacylglycerols, high-density lipoprotein cholesterol, and insulin. Insulin resistance was reduced, without affecting glucose levels and glucose tolerance. In contrast, PHE increased rectal temperature and slightly increased energy expenditure. The mitigation of HFD-induced metabolic disturbances points toward a promising role for PHE in obesity treatment and encourages further research on its mechanisms of action. SIGNIFICANCE STATEMENT: Phenelzine reduces body fat, markers of oxidative stress, inflammation, and insulin resistance in high-fat diet mice. Semicarbazide-sensitive amine oxidase, monoamine oxidase, phosphoenolpyruvate carboxykinase, and sterol regulatory element-binding protein-1c are involved in the metabolic effects of phenelzine. Phenelzine could be potentially used for the treatment of obesity-related complications.

Roux-en-Y Gastric Bypass Improves Metabolic Conditions in Association with Increased Serum Bile Acids Level and Hepatic Farnesoid X Receptor Expression in a T2DM Rat Model

Roux-en-Y gastric bypass (RYGB) is an effective surgical treatment for type 2 diabetes mellitus (T2DM). The present study aimed to investigate the effects of RYGB on glucose homeostasis, lipid metabolism, and liver morphological adaption, as well as the changes in bile acids signaling and expression of its target regulatory factors involved in gluconeogenesis, lipogenesis, and fatty acid β oxidation. Twenty adult male T2DM rats induced by high-fat diet and a low dose of streptozotocin were randomly divided into sham and RYGB groups. The parameters of body weight, food intake, glucose tolerance, insulin sensitivity, serum lipid profiles, and bile acids level were assessed to evaluate metabolic changes. Liver sections were stained with hematoxylin-eosin (H&E) and oil red O to assess lipid accumulation. The mRNA and protein expression levels of farnesoid X receptor (FXR), small heterodimer partner (SHP), key regulatory factors of gluconeogenesis, lipogenesis, and fatty acid β oxidation (phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-α (PPAR-α)) were determined through RT-PCR and Western blotting, respectively. RYGB induced significant improvements in glucose tolerance and insulin sensitivity, along with weight loss and decreased food intake. RYGB also decreased serum TG, FFAs, and increased bile acids levels. The lipid droplets in the liver were significantly decreased after RYGB. The RYGB group exhibited downregulated mRNA and protein expression levels of PEPCK, G6Pase, and SREBP-1c and upregulated expression of FXR, SHP, and PPAR-α in the liver. RYGB ameliorates glucose and lipid metabolism accompanied by weight loss and calorie restriction. The liver exhibited a marked improvement in lipid accumulation after RYGB. The bile acids level, FXR, and its target transcriptional factor SHP expression were elevated. Meanwhile, our study demonstrated that the increased bile acids-FXR signaling, followed by the reduced hepatic gluconeogenesis, lipogenesis, and increased fatty acid β oxidation may contribute to improved metabolic conditions after RYGB.

Maternal di-(2-ethylhexyl) phthalate exposure alters hepatic insulin signal transduction and glucoregulatory events in rat F1 male offspring.

Di-(2-ethylhexyl) phthalate (DEHP) is a commonly used plasticizer with endocrine disrupting properties. Its widespread use resulted in constant human exposure including fetal development and postnatal life. Epidemiological and experimental data have shown that DEHP has a negative influence on glucose homeostasis. However, the evidence regarding the effect of maternal DEHP exposure on hepatic glucose homeostasis is scarce. Hence, we investigated whether DEHP exposure during gestation and lactation disrupts glucose homeostasis in the rat F1 male offspring at adulthood. Pregnant rats were divided into three groups and administered with DEHP (10 and 100mg/kg/day) or olive oil from gestational day 9 to postnatal day 21 (lactation period) through oral gavage. DEHP-exposed offspring exhibited hyperglycemia, impaired glucose and insulin tolerances along with hyperinsulinemia at postnatal day 80. DEHP exposure significantly reduced the levels of insulin signaling molecules such as insulin receptors, IRS1, Akt and its phosphorylated forms. GSK3β and FoxO1 proteins increased in DEHP-exposed groups whereas its phosphorylated forms decreased. Treated groups showed decreased glycogen synthase activity and glycogen concentration. Glucose-6-phosphatase and phosphoenolpyruvate carboxykinase mRNA level and enzyme activity increased in DEHP-treated groups. The interaction between FoxO1-glucose-6-phosphatase and FoxO1-phosphoenolpyruvate carboxykinase was also increased. This study suggests that DEHP exposure impairs insulin signal transduction and alters glucoregulatory events leading to the development of type 2 diabetes in F1 male offspring.

Synergistic effects of dietary carbohydrate and taurine on growth performance, digestive enzyme activities and glucose metabolism in juvenile turbot Scophthalmus maximus L.

A 10-week feeding trial was conducted to evaluate the effects of taurine on growth performance, digestive enzyme activities and glucose metabolism of juvenile turbot Scophthalmus maximus (initial body weight, 3.66 ± 0.02 g) fed diets with different carbohydrate level. With a 2 × 3 two factoral experimental design, two levels of dietary carbohydrate (15% and 21%, respectively) with three levels of taurine (0.4%, 1.2%, and 2.0%, respectively) were used. In addition, the diet with 15% carbohydrate and without taurine supplementation was used as the control. The results showed that, compared with the control group, 1.2% of dietary taurine supplementation significantly increased the specific growth rate, the activities of amylase and lipase in intestine, glycogen content and the mRNA levels of glucokinase, phosphofructokinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (G6PD), glycogen synthase (GS) and glucose transporter 2 in liver at 15% of dietary carbohydrate level. Meanwhile, it significantly decreased the blood glucose concentration and the mRNA levels of cytosolic phosphoenolpyruvate carboxykinase (cPEPCK) in liver. In addition, significant interaction between dietary carbohydrate and taurine were observed in blood glucose level. Compared to 15% dietary carbohydrate, 21% dietary carbohydrate significantly increased the blood glucose level at 1.2% and 2.0% dietary taurine level, whereas it had no significant effect at 0.4% dietary taurine supplementation level. At 21% of dietary carbohydrate level, although 1.2% and 2.0% of dietary taurine supplementation had no significant effects on blood glucose level, it significantly decreased the advanced glycationend products level in plasma and the fructose-1, 6-bisphosphatase mRNA level in liver. Moreover, compared to that fed diet with 15% carbohydrate, fish fed diet with 21% carbohydrate had lower activities of amylase and lipase in intestine and lower mRNA levels of G6PD, mitochondrial phosphoenolpyruvate carboxykinase and cPEPCK in liver. Furthermore, significant interactions between dietary carbohydrate and taurine were also observed in the expression of the genes involved in liver glucose metabolism above in addition to GS and cPEPCK. Overall, this study indicated that taurine had a beneficial effect on glucose homeostasis through enhancement of glycolysis, glucose transport, glycogen synthesis and the inhibition of gluconeogenesis in liver of turbot, thus decreased blood glucose level. However, the beneficial effect of taurine on glucose metabolism was affected by dietary carbohydrate level.