Effect of BP on lipid contents in the HFD-induced obese rats.

<p>(A, B, C) Significant decreases in the levels of serum triglyceride and total cholesterol were observed in the BPE-treated groups compared with HFD-induced obese rats. HDL-cholesterol levels in the BP groups were increased compared with the HFD groups. The values are expressed as the means ± SD. Bars with different letters are significantly different (p<0.05) as determined by Duncan's multiple range test.</p

Effect of BP on phosphorylation of Akt and GSK3β in 3T3-L1 adipocytes.

<p>(A) Effect of BP on Akt activation in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with BP extracts at the indicated concentrations and the phosphorylation levels for Akt was determined by Western blotting analysis. The data are presented as the means ± SD values for at least three independent experiments. *P<0.05. (B) Effect of BP on GSK3β activation in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with BP extracts at the indicated concentrations, and the phosphorylation levels for GSK3β were determined by Western blotting analysis. The data are presented as the means ± SD values of at least three independent experiments. *P<0.05. (C) Effects of the PI3K/Akt inhibitor LY294002 (10 µM) on BP-induced inhibition of adipocyte differentiation in 3T3-L1 cells. 3T3-L1 cells were treated with BPE during differentiation in the presence or absence of the LY294002. The intracellular lipid accumulation was measured by triglyceride assay. Data are expressed as mean ± SD of three independent experiments. *P<0.05.</p

BPE inhibits intracellular lipid accumulation in 3T3-L1 cells.

<p>(A) Hormone-induced differentiation of 3T3-L1 adipocytes was repressed by BPE. Confluent 3T3-L1 preadipocytes differentiated into adipocytes in medium containing different concentrations of BPE for 7 days (from day 0 to 7). Oil-red O staining was performed on day 7. DMI: fully differentiated-adipocytes (0.5 mM 3 IBMX, 100 µM indomethacin, 0.25 µM dexamethasone and 167 nM insulin). BPE: blueberry peel extracts. (B) BPE reduced TG accumulation in differentiated 3T3-L1 cells. The data shown are representative of at least three independent experiments. The values are presented as the means ± SD. Bars with different letters are significantly different (p<0.05) as determined by Duncan's multiple range test. (C) The effect of BP on cell viability in preadipocytes. 3T3-L1 preadipocytes were incubated with BP extracts (0–300 µg/mL) for 7 days. Cell viability after treatment with BP was determined by the MTT assay. The values are presented as the means ± S.D. The data shown are representative of at least three independent experiments.</p

Effect of BP on the expression of adipogenic genes in 3T3-L1 adipocytes.

<p>3T3-L1 preadipocytes were differentiated into adipocytes in DMI medium in the absence or presence of 50 µg/mL or 200 µg/mL BPE for 4 or 7 days. (A) BPE inhibited the expression of adipocyte-specific transcription factors during differentiation. The gene expression analysis was performed by RT-PCR, and all of the gene transcripts were normalized using β-actin as a control. All of the experiments were performed in three independent experiments. Bars with different letters are significantly different (p<0.05) as determined by Duncan's multiple range test. (B) BP reduced the expression of adipogenesis-related genes in 3T3-L1 adipocytes. Total cell lysates were isolated from 3T3-L1 adipocytes at day 4 or day 7 after induction of differentiation. Western blotting analysis was performed as described in the Materials and Methods.</p

Effects of BP extracts on body weight in HFD-induced obese rats.

<p>(A) ND groups (▪) were fed normal diet (ND), HFD-SBP groups were fed HFD plus BPE (60 mg/kg BW, ▴), HFD-LBP groups (♦) were fed HFD plus BPE (150 mg/kg BW), and HFD groups (×) were fed high-fat diet. The body weight was measured twice a week. Body weights at the end of the experiments were significantly different between the HFD and ND (P<0.01) and HFD-BP groups (P<0.05). (B, C) BPE treatment decreased perirenal and epididymal fat weights in HFD-induced obese rats. The weights of the perirenal and epididymal fatty tissue were calculated by dividing the fatty tissue weight by body weight (fatty tissue/body weight x 100). The values are expressed as the means ± SD. Bars with different letters are significantly different (p<0.05) as determined by Duncan's multiple range test.</p

Antioxidant capacities, total phenolic and flavonoids content of BP extracts.

a<p>DPPH, DPPH radical scavenging activity;</p>b<p>HRSA, hydroxyl radial scavenging activity;</p>c<p>SRSA, superoxide anion radical scavenging activity;</p>d<p>TPC, total phenolic acid. Total phenolic acid and total flavonoid content expressed as milligrams of quercetin equivalent (QE)/g of extract.</p>1)<p>The positive controls of DPPH, HRSR and SRSA were ascorbic acid, ascorbic acid and quercetin, respectively.</p>x–z<p>The values are presented as the means ± SD. P<0.01 represents a significant difference between the samples (n = 4).</p

Additional file 1: of Rubus crataegifolius Bunge regulates adipogenesis through Akt and inhibits high-fat diet-induced obesity in rats

Effects of supplementing RCB on body weight gain, and serum profiles in rats fed a high fat diet for two weeks. (A) Weight gain, (B) serum triglyceride level, and (C) total cholesterol level. The values are expressed as the mean ± SD. The bars showing different letters indicate significant differences among each group of bars, according to Duncan’s test; *p < 0.05. (ZIP 90 kb

Determination of gut lesion scores, serum carotenoid levels, and IFN-γ transcript levels.

<p>Ten-day-old chickens were orally infected with 1×10⁴ sporulated <i>E. tenella</i> oocysts. Nine chickens were randomly chosen for serum samples and gut lesion scoring 7 days after <i>Eimeria</i> infection. (A) Lesion scores (0–4) were based on scoring techniques previously described (Johnson and Reid, 1970). (B) Serum samples were extracted with ten volumes of acetone to precipitate proteins. Absorbencies of the supernatants were determined spectrophotometrically at 456 nm using a β-carotene standard. Bars represent the means ± standard error from nine chickens. (C) Expression of IFN-γ mRNA in cecal-tonsils of chickens infected with <i>E. tenella</i>. Tissue samples were pooled from five chickens and subjected to quantitative real-time PCR. Expression levels were normalized to those of β-actin from the same samples. The <i>y</i> axis represents the fold change in expression of IFN-γ gene from <i>E. tenella</i>-infected chickens as compared to uninfected chickens. Data represent means ± standard error of triplicate samples. * <i>P</i><0.05, ** <i>P</i><0.01 or *** <i>P</i><0.001 was considered significant compared to uninfected chickens. Data are representative of two independent experiments with similar pattern results.</p

List of primers used in quantitative real-time PCR.

<p>List of primers used in quantitative real-time PCR.</p

Distribution and molecular weight of chicken IL2/15Rβ.

<p>(A) Expression of chIL2/15Rβ transcripts in various chicken tissues and cell lines. Total RNA was isolated from various tissues of 10-day-old chickens and analyzed with quantitative real-time PCR. Tissue samples were pooled from five chickens. Expression levels were normalized to those of β-actin from the same samples. Data represent means of triplicate samples. Data are representative of two independent experiments with similar pattern results. CU205, REV-transformed lymphoblast cell line; HD11, macrophage cell line; ND, not detected. (B) Detection of chicken IL2/15Rβ protein with Western blot analysis. Whole-cell lysates of COS-7 cells were collected 48 h (lanes 1 and 2) after transient transfection with a chIL2/15Rβ-HA construct (lane 2) or empty pcDNA 3.1 (lane 1). To determine the size of the chIL2/15Rβ backbone, transfected cells were incubated for 24 h and then treated with 5 µg/ml tunicamycin as an inhibitor of N-linked glycosylation followed by incubation for an additional 6 h (lane 3) and 24 h (lane 4). Cell lysates from COS-7 cells were separated by SDS-PAGE under reducing conditions. Arrows indicate specific bands. Data are representative of three independent experiments with similar pattern results.</p