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

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

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

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

Comparison of Soluble Aβ42/40 by the CG extracts on Tg mice brain.

<p>The cerebral Aβ42(A) and Aβ40 (B) load was quantitated by capturing antigen such as Amyloid beta 42 and/or Amyloid beta 40 mediated Enzyme-Linked Immunosorbent Assay (ELISA) kit (Invitrogen, CA) using brain lysates (i.e., total brain lysate of hippocampus and neocortex area) from the EA-CG-treated or vehicle (PBS)-treated Tg (Mo/Hu APPswe PS1dE9) mice. The cerebral Aβ42 concentrations in the EA-CG-treated mice were significantly reduced compared to the vehicle-treated controls (**P < 0.01). The values shown were the means ± S.E.M. (n = 10 per group).</p

Comparison of the amyloid burden in the Tg brains by the CG extracts.

<p>(A) The amyloid plaques were identified by immunohistochemistry (IH) and H&E counterstaining. The CG extract-treated (EA-CG fraction) and control Tg brain sections were formalin fixed and subjected to immunohistochemistry with the 6E10 antibody that interacts with the Aβ1–16 epitope (monoclonal antibody) in the hippocampus (a, control; b, EA-CG treated) and cortex (e, control; f, EA-CG-treated). Concomitant with the IH staining, other hippocampal (n = 6) (c, vehicle; d, EA-CG-treated) and cortical (n = 6) (g, vehicle; h, EA-CG-treated) sections from the Mo/Hu APPswe PS1dE9 mice were subjected to H&E counterstaining. The scale bar indicates 200 μm (a through h). The amyloid load (% of stained area) as a quantitation result was decreased by the CG extracts (B. hippocampus area; C. cortex area). The area in the hippocampus (D, vehicle alone vs. EA-CG) and cortex (E, vehicle alone vs. EA-CG) was quantified by comparing the morphometric analysis of the Thioflavin-S-stained area between the vehicle-treated vs. EA-CG-treated Mo/Hu APPswe PS1dE9 brain sections. The values (%) shown were the means ± S.E.M. *p < 0.05, Tg vs. Tg+EA-CG.</p

Effect of CG extracts on liver toxicity using aspartate/alanine aminotransferase activity in Tg mice.

<p>The Mo/Hu APPswe PS1dE9 mice were fed with or without EA-CG (50 mg/kg BW) for 6 months. The brain were isolated from the EA-CG treated and control (PBS) groups (n = 3 mice per each group). After the brain was divided in half, we tested the gene expression profile of the inflammatory cytokines IL-1α (A) and IFN-γ (B) as markers of the cellular response by semi-quantitative RT-PCR. The fold change was presented after normalization to the housekeeping gene β-actin). *p < 0.05; n.s. mean not significant. The levels of Aspartate aminotransferase activity (AST, Units/L) and Alanine aminotransferase activity (ALT, Units/L)) were measured in the sera by centrifugation after obtain mice blood from each group, and then the relative value (U/L) of ALT (C) or AST (D) as hepatotoxicity markers were measured and presented after comparison to the controls (PBS groups), according to the manufacturer’s instructions. The values were presented as the means ± S.E.M (n = 3 mice per each group). n.s. mean not significant.</p