Carob pod polyphenols suppress the differentiation of adipocytes through posttranscriptional regulation of C/EBPβ.

Obesity is a major risk factor for various chronic diseases such as diabetes, cardiovascular disease, and cancer; hence, there is an urgent need for an effective strategy to prevent this disorder. Currently, the anti-obesity effects of food ingredients are drawing attention. Therefore, we focused on carob, which has high antioxidant capacity and various physiological effects, and examined its anti-obesity effect. Carob is cultivated in the Mediterranean region, and its roasted powder is used as a substitute for cocoa powder. We investigated the effect of carob pod polyphenols (CPPs) on suppressing increases in adipose tissue weight and adipocyte hypertrophy in high fat diet-induced obesity model mice, and the mechanism by which CPPs inhibit the differentiation of 3T3-L1 preadipocytes into adipocytes in vitro. In an in vivo experimental system, we revealed that CPPs significantly suppressed the increase in adipose tissue weight and adipocyte hypertrophy. Moreover, in an in vitro experimental system, CPPs acted at the early stage of differentiation of 3T3-L1 preadipocytes and suppressed cell proliferation because of differentiation induction. They also suppressed the expression of transcription factors involved in adipocyte differentiation, thereby reducing triacylglycerol synthesis ability and triglycerol (TG) accumulation. Notably, CPPs regulated CCAAT/enhancer binding protein (C/EBP)β, which is expressed at the early stage of differentiation, at the posttranscriptional level. These results demonstrate that CPPs suppress the differentiation of adipocytes through the posttranscriptional regulation of C/EBPβ and may serve as an effective anti-obesity compound

Ultrasound microbubble-mediated transfection of NF-κB decoy oligodeoxynucleotide into gingival tissues inhibits periodontitis in rats in vivo

<div><p>Periodontitis is a chronic infectious disease for which the fundamental treatment is to reduce the load of subgingival pathogenic bacteria by debridement. However, previous investigators attempted to implement a nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotide (ODN) as a suppressor of periodontitis progression. Although we recently reported the effectiveness of the ultrasound-microbubble method as a tool for transfecting the NF-κB decoy ODN into healthy rodent gingival tissue, this technique has not yet been applied to the pathological gingiva of periodontitis animal models. Therefore, the aim of this study was to investigate the effectiveness of the technique in transfecting the NF-κB decoy ODN into rats with ligature-induced periodontitis. Micro computed tomography (micro-CT) analysis demonstrated a significant reduction in alveolar bone loss following treatment with the NF-κB decoy ODN in the experimental group. RT-PCR showed that NF-κB decoy ODN treatment resulted in significantly reduced expression of inflammatory cytokine transcripts within rat gingival tissues. Thus, we established a transcutaneous transfection model of NF-κB decoy ODN treatment of periodontal tissues using the ultrasound-microbubble technique. Our findings suggest that the NF-κB decoy ODN could be used as a significant suppressor of gingival inflammation and periodontal disease progression.</p></div

Alveolar bone loss in control, periodontitis (P), and periodontitis with ultrasound-microbubble-mediated application of NF-κB decoy oligodeoxynucleotide (ODN) (PUM) in rats.

<p>Graphic depictions of the mean alveolar bone loss (mm) between <b>(A)</b> the first (M1) and second (M2) maxillary molars and <b>(B)</b> M2 and the third maxillary molar (M3) in the rats in each group. Results are expressed as the mean ± standard error (n = 6) of the distance from the cement enamel junction (CEJ) to the alveolar bone crest (ABC. *P < 0.05; N.S., not significant).</p

Histological images of the P and PUM groups.

<p>Comparative HE staining images taken at the M2 of representative rats in the P and PUM groups. The images on the right are magnified views of the inset in the corresponding left images. <b>(A)</b> Schematic illustration of these coronal sections in P and PUM groups. The inset in the schematic illustration indicates the area of the B images. (<b>B- a, b</b>) The periodontium of the P group rats showed increases in the capillaries (black arrows) and thickening of the junctional epithelium (yellow arrows). (<b>B- c, d</b>) The periodontium of the PUM rats showed decreased inflammatory response compared to the P group. Lig: silk ligature, E: junctional epithelium, C: gingival connective tissue. Bar = 100 μm.</p

Three-dimensional analysis.

<p>(<b>A, B and C</b>) Three-dimensional images of rats with periodontitis (P); A: buccal side view, B: occlusal surface view, C: palatal side view. (<b>D, E and F</b>) Three-dimensional images of periodontitis with ultrasound-microbubble-mediated application of NF-κB decoy oligodeoxynucleotide (ODN) (PUM) in rats; D: buccal side view, E: occlusal surface view, F: palatal side view. <b>(G)</b> Alveolar bone volume in control, P, and PUM rats. Graphic depiction of alveolar bone volume (mm³) in the rats in each group. Results are expressed as the mean ± standard error (n = 6) of the volume of alveolar bone. *P < 0.05.</p

Depiction of the method used for micro-computed tomography (micro-CT) analysis of rat molars.

<p><b>(A)</b> Linear measurements were taken of alveolar bone loss in the interdental space from the cement enamel junction (CEJ) to the alveolar bone crest (ABC). <b>(B)</b> Representative depiction of the region of interest (ROI), which comprised a rectangular area drawn 0.33 mm axially from the tooth crown in each direction. <b>(C)</b> Representative ROI including, vertically, the coronal halves of the slices through the mesial root apex to the CEJ. <b>(D and E)</b> Representative three-dimensional (3D) images of the volume of interest (VOI): <b>(D)</b> horizontal view <b>(E)</b> sagittal view. <b>(F)</b> Representative volumetric measurement obtained using the 3D-generated ROI. Colored regions (blue: mesial palatal root of M2, orange: distal palatal root of M2, red: mesial buccal root of M2, yellow-green: distal buccal root of M2, light blue and yellow: distal root of M1, green: mesial root of M3) indicate tooth parts that were removed from the VOI for measurement of the alveolar bone volume.</p

Evaluation of the expression of periodontitis markers of palatal gingiva in control, periodontitis (P), and periodontitis with ultrasound-microbubble-mediated application of NF-κB decoy oligodeoxynucleotide (ODN) (PUM) in rats.

<p>Relative expression levels of <i>IL1B</i>, <i>TNFα</i>, <i>ICAM1</i>, and <i>RANKL</i> within the palatal gingival tissues of rats in each group (n = 6 per group) were determined by RT-PCR analysis at <b>(A)</b> 7 days and <b>(B)</b> 14 days post-treatment. The mRNA expression levels in the control group were set to a value of 1. Data are presented as the mean ± standard deviation for each group. *P < 0.05.</p