Oleic Acid Promotes the Biosynthesis of 10-Hydroxy-2-decenoic Acid via Species-Selective Remodeling of TAGs in Apis mellifera ligustica.

This study aimed to assess the impact of oleic acid (OA) supplementation on the biosynthesis of 10-hydroxy-2-decenoic acid (10-HDA) in Apis mellifera ligustica. In experiment 1, varying concentrations of OA (2%, 4%, 6% and 8%) were added to an artificial diet for newly emerged bees reared in cages. Analysis of 10-HDA content and gene expression in the mandibular gland (MG) revealed that the 8% OA treatment had the greatest impact on promoting the synthesis of 10-HDA. Subsequent investigations utilized RNA-seq and lipidomics to characterize the molecular signature in the MG after feeding the 8% OA diet. Phosphatidylcholine (PC) and triacylglycerol (TAG) were found to be the predominant lipids in the MG of worker bees. A total of 154 TAGs were identified, with TAG (18:1-18:1-18:1) exhibiting the highest abundance, which increased by 1.5 times. The major TAG species contained palmitic acid (16:0) and oleic acid (18:1) in their structure, which was associated with fatty acid composition of diet. The increase in abundance of main TAGs may be attributed to the upregulation of glycerol-3-phosphate acyltransferase (Gpat) and glycerol kinase (GK) gene expression at the transcriptional level. The upregulation of differentially expressed genes (DEGs) related to carbohydrate metabolism may contribute to meeting the heightened metabolic demands of the MGs in worker bees. Royal jelly (RJ) samples from bee colonies fed with the 8% OA diet exhibited higher 10-HDA level than RJ collected from bee colonies fed with the artificial diet. These results indicate that 8% OA addition in the diet enhanced biosynthesis of 10-HDA in the mandibular gland, which was accompanied by significant and highly species-selective remodeling of TAGs.

Selenium-rich royal jelly inhibits hepatocellular carcinoma through PI3K/AKT and VEGF pathways in H22 tumor-bearing mice.

Royal jelly (RJ) and selenium (Se)-rich foods have well-known health benefits that are attributable to a broad range of pharmacological effects including antioxidant, anti-tumor, and immunoregulatory activities. However, the physiological effects of Se-rich RJ, which is produced by feeding Apis mellifera (Hymenoptera: Apidae) sodium selenite sucrose solution, are not well understood. The anti-hepatoma activity and mechanism of Se-rich RJ in H22 tumor-bearing mice were investigated in the current study. The findings showed that the content of organic and inorganic Se in Se-rich RJ was significantly higher than that in RJ. Furthermore, interleukin-2 (IL-2) levels and tumor necrosis factor-α (TNF-α) production in serum were increased and the malondialdehyde (MDA) content in liver was decreased in mice fed RJ and Se-rich RJ. 16SrRNA sequencing and serum untargeted metabolomics showed that RJ and Se-rich RJ could modulate the gut microbiota, and fisetin and L-glutathione oxidized were the main anti-tumor components in RJ and Se-rich RJ. Further analysis showed 11-deoxy prostaglandin F1ß was the specific anti-tumor metabolite in mice treated with Se-rich RJ compared with RJ. The results indicated that RJ and Se-rich RJ could inhibit the expression of PI3K and phosphorylation of AKT, induce cell apoptosis through the activation of caspase-9 and caspase-3, and regulate Bcl-2/Bax expression. RJ and Se-rich RJ also inhibited the expression of COX-2 and VEGF. To summarize, the findings clearly demonstrate that Se-rich RJ could inhibit tumor growth by inducing apoptosis and inhibiting angiogenesis as well as exhibit anti-tumor effects by improving immune function and antioxidant activities. The results indicated that Se-rich RJ could be a potential functional food for the management and prevention of cancer.

The AMPK-mTOR signaling pathway is involved in regulation of food intake in the hypothalamus of stressed chickens.

Glucocorticoids (GCs) can stimulate the appetite and AMPK in broilers. The activation of hypothalamic mTOR has been proposed as an important anorexigenic signal. However, inhibitory effect of AMPK activity on appetite and AMPK downstream signaling pathway under stress has not been reported. In this study, we performed an intracerebroventricular (icv) injection of compound C, an AMPK inhibitor, in GC-treated birds to explore the regulatory mechanism on appetite and AMPK downstream signaling pathway. A total of 48 7-day-old broilers, which had received an icv cannula, were randomly subjected to one of two treatments: subcutaneous injection of dexamethasone (DEX) or saline. After 3 days of continuous DEX injection, chicks of each group received an icv injection with either compound C (6 µg/2 µL) or vehicle (dimethyl sulfoxide, 2 µL). The results showed that body weight gain was reduced by the DEX treatment. Compared with the control, icv injection of compound C reduced feed intake at 0.5-1.5 h. In the DEX-treated group, the inhibitory effect of compound C on appetite remained apparent at 0.5-1 h. The DEX treatment increased the gene expression of liver kinase B1 (LKB1), neuropeptide Y (NPY), and decreased p-mTOR protein level. In stressed broilers, inhibition of AMPK relieved the decreased mTOR activity. A significant interaction was noted in DEX and compound C on protein expression of phospho-AMPK. Taken together, in stressed broilers, the central injection of compound C could inhibit central AMPK activity and reduce appetite, in which the AMPK/mTOR signaling pathway might be involved.

The mixed application of organic and inorganic selenium shows better effects on incubation and progeny parameters.

This experiment aims to study the effects of dietary selenium (Se) sources on the production performance, reproductive performance, and maternal effect of breeder laying hens. A total of 2,112 Hyline brown breeder laying hens of 42 wk of age were selected and randomly divided into 3 groups, with 8 repeats in each group and 88 chickens per repeat. The sources of dietary Se were sodium selenite (SS, added at 0.3 mg/kg), L-selenomethionine (L-SM, added at 0.2 mg/kg), and combination of SS and L-SM (SS 0.15 mg/kg + L-SM 0.15 mg/kg). The pretest period was 7 d, and the breeding period was 49 d. Compared with 0.3 mg/kg SS, the addition of 0.2 mg/kg L-SM in the diet significantly increased the hatchability (P < 0.05) and the Se content (P < 0.05) in egg yolk and chicken embryo tissues and improved the activity of yolk glutathione peroxidase (GSH-px) effectively (P < 0.05). Treatment with 0.2 mg/kg L-SM also reduced the content of yolk malondialdehyde (P < 0.05) and significantly improved the antioxidant performance of 1-day-old chicks, as manifested by increased activity of antioxidant enzymes (GSH-px, total antioxidant capacity and the ability to inhibit hydroxyl radicals) in serum, pectoral, heart, and liver (P < 0.05). This treatment decreased the malondialdehyde content (P < 0.05) and increased the expression of liver glutathione peroxidase 4 and deiodinase 1 mRNA (P < 0.05). Adding L-SM to the diets of chickens increased the hatchability of breeder eggs as well as the amount of Se deposited and antioxidant enzyme activity in breeder eggs and embryos. Compared with SS, L-SM was more effectively transferred from the mother to the embryo and offspring, showing efficient maternal nutrition. For breeder diets, the combination of organic and inorganic Se (0.15 mg/kg SS + 0.15 mg/kg L-SM) is an effective nutrient supplementation technology program for effectively improving the breeding performance of breeders and the antioxidant performance and health level of offspring chicks.

Effects of dietary corticosterone on the central adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in broiler chickens.

Glucocorticoids (GCs) induce the activation of the central adenosine 5'-monophosphate-activated protein kinase (AMPK) signaling pathway in birds. In this study, we aimed to investigate the effects of corticosterone (CORT) supplemented in diet on the central AMPK signaling pathway in broilers. The average daily gain was reduced by CORT treatment, and the average daily feed intake remained unchanged. Plasma glucose, triglyceride, total cholesterol, and CORT contents were increased by CORT administration. In addition, CORT treatment decreased the relative weights of heart, spleen, and bursa and increased the relative weights of liver and abdominal fat. The glycogen contents in the liver and breast muscle were higher in the chicks treated with CORT. CORT treatment upregulated the gene expression of mammalian target of rapamycin, glucocorticoid receptor, AMPKα2, neuropeptide Y(NPY), liver kinase B1 (LKB1), AMPKα1, and fatty acid synthase in the hypothalamus. Moreover, CORT treatment increased the protein levels of acetyl-coenzyme A carboxylase (ACC) phosphorylation and total AMPK and phosphorylated AMPK in the hypothalamus. Hence, CORT administration in the diet activated the LKB1-AMPK-NPY/ACC signaling pathway in the hypothalamus of broiler.

Effects of dietary energy level on appetite and central adenosine monophosphate-activated protein kinase (AMPK) in broilers.

Adenosine monophosphate-activated protein kinase (AMPK) acts as a sensor of cellular energy changes and is involved in the control of food intake. A total of 216 1-d-old broilers were randomly allotted into 3 treatments with 6 replicates per treatment and 12 broilers in each cage. The dietary treatments included 1) high-energy (HE) diet (3,500 kcal/kg), 2) normal-energy (NE) diet (3,200 kcal/kg), and 3) low-energy (LE) diet (2,900 kcal/kg). The present study was conducted to investigate the effects of dietary energy level on appetite and the central AMPK signal pathway. The results showed that a HE diet increased average daily gain (ADG), whereas a LE diet had the opposite effect (P < 0.05, N = 6). The average daily feed intake (ADFI) of the chickens fed the LE diet was significantly higher than that of the control (P < 0.05, N = 6). Overall, the feed conversion rate gradually decreased with increasing dietary energy level (P < 0.05, N = 6). Moreover, the chickens fed the LE and HE diets demonstrated markedly improved urea content compared with the control group (P < 0.0001, N = 8). The triglyceride (TG) content in the LE group was obviously higher than that in the HE group but showed no change compared with the control (P = 0.0678, N = 8). The abdominal fat rate gradually increased with increased dietary energy level (P = 0.0927, N = 8). The HE group showed downregulated gene expression levels of liver kinase B1 (LKB1), neuropeptide Y (NPY), cholecystokinin (CCK), and glucocorticoid receptor (GR) in the hypothalamus compared with the control group (P < 0.05, N = 8). However, LE treatment significantly increased the mRNA level of AMP-activated protein kinase α2 (AMPKα2) compared with other groups (P = 0.0110, N = 8). In conclusion, a HE diet inhibited appetite and central AMPK signaling. In contrast, a LE diet activated central AMPK and appetite. Overall, the central AMPK signal pathway and appetite were modulated in accordance with the energy level in the diet to regulate nutritional status and maintain energy homeostasis in birds.

Effect of Zinc on Appetite Regulatory Peptides in the Hypothalamus of Salmonella-Challenged Broiler Chickens.

The effects of dietary Zinc (Zn) supplementation on the gene expression of appetite regulatory peptides were investigated in Salmonella-infected broiler chickens. Broiler chickens (Arbor Acres, 1 day old) were allocated randomly into 24 pens of 10 birds. The chickens from 12 pens were fed with basal diet and the other with basal diet supplemented with Zn (ZnSO4·H2O, 120 mg/kg). At 5 days of age, the chickens were divided into 4 treatments with 6 pens: basal diet; basal diet and Salmonella challenge; Zn-supplemented diet; Zn-supplemented diet and Salmonella challenge. At 42 days of age, the hypothalamus from 6 chickens per treatment (1 chicken per pen) was individually collected for gene expression determination. Results showed that dietary supplementation of Zn reduced the gene expression of hypothalamic ghrelin and tumor necrosis factor alpha (TNF-α) (P < 0.05). Salmonella infection upregulated the messenger RNA (mRNA) levels of hypothalamic neuropeptide Y (NPY) and TNF-α. Zn supplementation and Salmonella inoculation were significantly correlated with the mRNA levels of toll-like receptor 2-1 (P < 0.05). However, neither dietary Zn supplementation nor Salmonella inoculation had significant effect on hypothalamic agouti-related protein, cocaine- and amphetamine-regulated transcript, and pro-opiomelanocortin. This study shows that dietary Zn supplementation promoted orexigenic appetite regulatory peptides and reduced the expression of the inflammatory cytokine TNF-α in the hypothalamus of Salmonella-challenged broilers.