Effects of Chinese Herbal Medicine Feed Additives on Growth Performance and Dietary Nutrient Metabolism of Black-bone Chickens

[Objectives] To study the effects of Chinese herbal medicine feed additives on the growth performance and dietary nutrient metabolic rate of black chickens. [Methods] 480 pieces of 1-day-old Donglan Black-bone Chickens were selected and divided into 4 groups, with 6 replicates in each group, with 20 pieces in each replicate. Among them, group A was the control group and was fed the basal diet; the groups B, C, and D were the experimental groups, which were separately supplemented with 0.25%, 0.50% and 1.00% of Chinese herbal medicine additives in the basal diet, and the experimental period was 70 d. At the end of the feeding experiment, 24 experimental chickens with similar body weight were selected from each group, and the same design of the feeding experiment was used to continue to feed the original diet for the metabolic experiment to determine the nutrient metabolic rate. [Results] Except the 50-70 d stage, the average daily gain (ADG) of group B in other stages was increased by 5.00-9.15% (P0.05); at 36-49 d, the average daily feed intake (ADFI) of group A was higher than that of group B by 6.04% (P0.05); at 22-35 and 1-70 d, the feed to gain ratio (F/G) of group B was 6.32% and 3.41% lower than that of group A (P<0.05), and at 36-49 and 1-49 d, the F/G of group B was even lower; at 36-49 d, the F/G of group C was 10.15% lower than that of group A (P<0.05), and at 1-49 d, it was also lower than that of group A by 5.29% (P<0.01); there was no significant difference between the two groups at other stages. In terms of nutrient metabolism: the three different amounts of this additive have increased the metabolic rate of DM, CP, CF, NFE, CA, Ca, P, TE and other nutrients in the diet to varying degrees, especially the addition of 0.25% and 0.50% had the significant increase. [Conclusions] The addition of 0.25% Chinese herbal medicine additives in the diet can significantly improve the growth performance and main nutrient metabolic rate of Donglan Black-bone Chickens

Insulin Stimulates Goose Liver Cell Growth by Activating PI3K-AKT-mTOR Signal Pathway

Background/Aims: Recent studies have suggested a crucial role for PI3K-Akt-mTOR pathway in regulating cell proliferation, so we hypothesize that insulin acts goose hepatocellular growth by PI3K-Akt-mTOR signal pathway. Because the physiological status of liver cells in vitro is different from that in vivo, a simplified cell model in vitro was established. Methods: Goose primary hepatocytes were isolated and incubated in either no addition as a control or insulin or PI3K-Akt-mTOR pathway inhibitors or co-treatment with glucose and PI3K-Akt-mTOR pathway inhibitors; Then, cell DNA synthesis and cell cycle analysis were detected by BrdU-incorporation Assay and Flow cytometric analysis; the mRNA expression and protein expression of factors involved in the cell cycle were determined by Real-Time RT-PCR, ELISA, and western blot. Results: Here we first showed that insulin evidently increased the cell DNA synthesis, the mRNA level and protein content of factors involved in the cell proliferation of goose primary hepatocytes. Meanwhile, insulin evidently increased the mRNA level and protein content of factors involved in PI3K-Akt-mTOR pathway. However, the up-regulation of insulin on cell proliferation was decreased significantly by the inhibitors of PBK-Akt-mTOR pathway, LY294002, rapamycin or NVP-BEZ235. Conclusion: These findings suggest that PI3K-Akt-mTOR pathway plays an essential role in insulin-regulated cell proliferation of goose hepatocyte

Treatment withLY294002 blocked the effect of insulin on lipid accumulation.

<p><b>A</b>, intracellular TG concentrations. <b>B</b>, extracellular TG concentrations. <b>C</b>, extracellular VLDL concentrations. <b>D</b>, Lipid contents were measured by Oil Red O extraction and are shown in optical density value units. E, Protein levels of genes involved in lipid metabolism; FAS levels are shown in nmol/ml, whereas ACCα and CPT1 levels are shown in ng/ml. * indicate significant differences among treatments (P <0.05).</p

Effect of insulin on protein levels and mRNA levels of genes involved in lipid metabolism.

<p><b>A</b>, Protein levels of genes involved in lipid metabolism; FAS levels are shown in nmol/ml, whereas ACCα and CPT1 levels are shown in ng/ml. <b>B</b>, Relative mRNA levels of genes related to lipogenesis. <b>C</b>, Relative mRNA levels of genes involved in fatty acid oxidation. <b>D</b>, Relative mRNA levels of genes that participate in VLDL-TG assembly and secretion. * indicate significant differences among treatments (P<0.05).</p

Treatment with NVP-BEZ235 affected the regulation of insulin on mRNA levels of genes involved in lipid metabolism.

<p><b>A</b>, Relative mRNA levels of genes related to lipogenesis. <b>B</b>, Relative mRNA levels of genes involved in fatty acid oxidation. <b>C</b>, Relative mRNA levels of genes that participate in VLDL-TG assembly and secretion. * indicate significant differences among treatments (P<0.05).</p

Treatment with LY294002, rapamycin, or NVP-BEZ235 decreased the insulin-stimulated up-regulation of ACCα protein expression.

<p>The concentrations of insulin, LY294002, rapamycin, and NVP-BEZ235 were 150 nmol/L insulin, 20 μmol/L, 30 nmol/L, and 1 μmol/L NVP-BEZ235 respectively. The blots are representative of three independent experiments.</p

Treatment with LY294002 affected the regulation of insulin on mRNA levels of genes involved in lipid metabolism.

<p><b>A</b>, Relative mRNA levels of genes related to lipogenesis. <b>B</b>, Relative mRNA levels of genes involved in fatty acid oxidation. <b>C</b>, Relative mRNA levels of genes that participate in VLDL-TG assembly and secretion.* indicate significant differences among treatments (P<0.05).</p

Primer sequences for real-time PCR.

<p>Primer sequences for real-time PCR.</p

Treatment with rapamycin blocked the effect of insulin on lipid accumulation.

<p><b>A</b>, intracellular TG concentrations. <b>B</b>, extracellular TG concentrations. <b>C</b>, extracellular VLDL concentrations. <b>D</b>, Lipid contents were measured by Oil Red O extraction and are shown in optical density value units. <b>E</b>, Protein levels of genes involved in lipid metabolism; FAS levels are shown in nmol/ml, whereas ACCα and CPT1 levels are shown in ng/ml. * indicate significant differences among treatments (P <0.05).</p