Effect of TAK-937 on rectal and brain temperature in rats with transient MCAO.

<p>Effect of TAK-937 on rectal and brain temperature in rats with transient MCAO.</p

Contribution of hypothermia to cerebroprotective effects of TAK-937 after transient MCAO in rats.

<p>Brain temperature (<b>A</b>) and infarct volume (<b>B</b>). Data are indicated as the means ± SEM. Significant differences from the corresponding vehicle-treated group are indicated by ^***<i>P</i><0.001 and ^*<i>P</i><0.05 (Dunnett’s test), and from the corresponding TAK-937-treated group is indicated by ^##<i>P</i><0.01 (Student’s <i>t</i> test). Numbers of rats used are shown in parentheses.</p

Reversal of cerebroprotection of TAK-937 by AM251, a CB₁ antagonist, after transient MCAO in rats.

<p>Rectal temperature (<b>A</b>) and infarct volume (<b>B</b>). Data are indicated as the means ± SEM. Significant differences from the corresponding vehicle-treated group are indicated by^***<i>P</i><0.001 (Dunnett’s test), and from the corresponding TAK-937-treated group is indicated by ^###<i>P</i><0.001 (Student’s <i>t</i> test). Numbers of rats used are shown in parentheses.</p

Canolol Inhibits Gastric Tumors Initiation and Progression through COX-2/PGE2 Pathway in K19-C2mE Transgenic Mice

<div><p>4-vinyl-2, 6-dimethoxyphenol (canolol) is an antioxidant phenolic compound extracted from crude canola oil. In current research, K19-C2mE transgenic mice, developing hyperplastic tumors spontaneously in the glandular stomach, were used to study the mechanisms involved in the anti-inflammation and anti-tumor effects of canolol. Tg mice receiving canolol diet had a reduced tumor incidence, to 41.2%, compared with Non-treatment Tg mice, 77.8% of which had gastric tumor (P=0.002). Besides that, the mean tumor diameter was decreased from 6.5mm to 4.5mm (P<0.001) after canolol administration. COX-2/PGE2 pathway is known to play pivotal role in inflammation-induced gastric tumorigenesis. The neutrophils and lymphocytes infiltration was suppressed significantly, and the mRNA levels of the proinflammatory cytokines COX-2, IL-1β and IL-12b were also downregulated in gastric mucosa. Additionally, immunohistochemical analysis showed that COX-2, EP2, Gαs and β-catenin, key factors involving in PGE2 signal transduction, were positive staining with higher H scores in Non-treatment Tg mice, while the expressions were suppressed significantly by 0.1% canolol (P<0.001). In addition, tumor-suppressor miR-7 was reactivated after canolol administration, and COX-2 was showed to be a functional target of miR-7 to suppress the tumor progression. In conclusion, canolol could inhibit the gastritis-related tumor initiation and progression, and the suppression effect was correlated with the blocking up of canonical COX-2/PGE2 signaling pathway and might be regulated by miR-7.</p></div

The effects of canolol on the COX-2/PGE2 pathway genes (A) and miR-7 expression (B).

<p>The studied genes involved proinflammatory cytokines (IL-12b, IL-1β, IL-6), oxidative responding gene HO-1, PGE2 synthases genes (COX-2, mPGES-1) and Gαs that relays PGE2 signal into cytoplasm. The horizontal bar in B indicates the 1.0 *, P<0.05; * *, P<0.01.</p

Experiment design in this study.

<p>4 weeks old mice were subjected to genotyping PCR, after identified, 6 weeks old Tg mice were randomly divided into Non-treatment group and Canolol-treated group. All the mice were fed with modified AIN93G diet with(out) 0.1% canolol till 52 weeks.</p

Histological examination showed the grades of gastritis.

<p>The normal gastric epithelial in wild type mice (grade 0, A), moderate (grade 2, C) and marked (grade 3, D) gastritis in Non-treatment Tg mice, and the alleviation of inflammatory responses in Canolol-treated Tg mice (grade 1, B). The arrow indicates mononuclear cell infiltration into the gastric glands or lymphoid follicle formed in submucosa. HE×20</p

Immunohistochemical staining of COX-2 (A, B), EP2 (C, D), Gαs (E, F) and β-catenin (G, H) in Non-treatment mice (left panel) and the Canolol-treated mice (right panel).

<p>Note that the intensity of COX-2, EP2, Gαs and β-catenin immunoreactivity in the Canolol-treated Tg gastric mucosa is weaker than that in Non-treatment Tg group. IHC×40.</p

COX-2 is the direct target of miR-7.

<p>The relative luciferase activity was reduced significantly in 3’-UTR-1 but not in 3’-UTR-2 (A). miR-7 and its putative two binding sequence in the 3’-UTR of COX-2, 86–92 sites (B) and 1111–1118 sites (C). The mutants of 3’-UTR were indicated by the arrows. *, P<0.05</p

The β-catenin expression in cytoplasm and nuclear in Non-treatment (1&3) and Canolol-treated (2&4) gastric tumor tissue.

<p>Note the β-catenin nuclear accumulation was decreased in Canolol-treated groups (Lane 4) compare to Non-treatment groups (Lane 3).</p