Influência da administração precoce de INFLIXIMAB na cicatrização de anastomose no cólon esquerdo de ratos com ou sem colite induzida

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Medicina, Programa de Pós-Graduação em Ciências Médicas, 2017.Objetivo: Avaliar a influência da utilização precoce do INFLIXIMAB sobre a cicatrização de anastomose no cólon esquerdo de ratos em um modelo experimental de colite, comparando a força de ruptura da anastomose e analisar a relação com a cicatrização da parede abdominal. Método: Trinta e dois ratos foram distribuídos em quatro grupos contendo 08 animais cada. Dois grupos (16 animais) foram submeditos a indução de colite por enema de ácido acético com dose de 3 ml por via retal, e dois grupos (16 animais) não sofreram indução de colite. Todos os ratos foram submetidos à laparotomia para exposição do cólon distal com secção do mesmo a cerca de 2,5 a 3,5 cm cranial à reflexão peritoneal e anastomose término-terminal do segmento. Tanto no grupo com colite induzida como no grupo sem colite, metade dos animais (8) receberam infliximab (IFX), 4 no 1 DPO e 4 no 3 DPO; e 8 receberam NaCl a 0,9%. No sétimo DPO foi realizada a pesagem dos animais e a re-laparotomia para colectomia, seguida de eutanásia. Foram avaliados a variação de peso, a força tensil de ruptura da anastomose e da parede abdominal, além de achados histopatológicos nas lâminas de anastomoses. Resultados: Nos animais com colite houve maior perda de peso em relação aos sem colite, mais acentuada nos que receberam IFX no 1o DPO (p=0,007). O IFX piorou a força tênsil de rutura da anastomose nos animais com colite quando administrado no 1o DPO (p=0,001), porém quando administrado no 3o DPO ou com placebo, o IFX melhorou a força de ruptura anastomótica nos animais com colite, sendo esta, maior do que os animais sem colite (p=0,001). Conclusão: O IFX prejudicou a cicatrização das anastomoses no cólon esquerdo de ratos com colite, quando administrado no 1o DPO. Quando administrado no 3o DPO ou com placebo houve uma inversão, com melhora nos valores de força de ruptura da anastomose em relação ao grupo sem colite.Objective: To evaluate the influence of the early use of INFLIXIMAB (IFX) on the anastomosis healing in the left colon of rats in an experimental model of acetic acid colitis induced, comparing the tensile strength of the anastomosis, and to analyze the relationship with the healing of the abdominal wall. Method: Thirty-two rats were divided into four groups containing 08 animals each. Two groups (16 rats) were carried out the induction of colitis by enema of acetic acid with dose of 3 ml, per rectally, with no induction of colitis in the other two groups (16 rats). All the rats were submitted to laparotomy for exposure of the distal colon and sectionated approximately 2.5 to 3.5 cm cranial from the peritoneal reflection and proceeded end-to-end anastomosis. In both groups, with and without colitis, half the animals (8) received infliximab administration,4 rats in the 1th POD and 4 in the 3th POD, and the other half (8) received NaCl solution0,9% administration. In the 7th POD the animals were weighed and relaparotomy was performed for colectomy followed by euthanasia. Were evaluated the weight variation, anastomosis and abdominal wall scar rupture tensile strength and histopathological findings by H-E stain. Results: In the animals with colitis, there was greater weight loss in relation to the without colitis group, more pronounced in who received IFX in the 1st POD (p=0.007). The IFX has deteriorated the rupture tensile strength of the anastomosis in the animals with colitis when administered in the 1st POD (p=0.001), but when administered in the 3rd POD or placebo, IFX has improved the anastomotic rupture tensile strength in animals with colitis, this being, greater than the animals without colitis (p=0.001). Conclusion: The Infliximab impaired the healing of anastomoses in the left colon of rats with colitis when administered in the 1st POD. When administered in the 3rd POD or when placebo was given, there was a reversal, with the improvement in the values of the breaking strength of the anastomosis in relation to the group without colitis

Additional file 3: Figure S3. of Kaiso differentially regulates components of the Notch signaling pathway in intestinal cells

CpG island prediction of the minimal JAG1 promoter. The JAG1 gene spanning −200 to +2500 bp of the TSS was analyzed for putative CpG islands. This GC-rich region harbors 5 potential CpG islands. (PDF 51 kb

Correlation of cytoplasmic Kaiso with clinicopathological features in invasive breast cancer.

<p>Correlation of cytoplasmic Kaiso with clinicopathological features in invasive breast cancer.</p

Correlation of nuclear Kaiso with the molecular subtypes of breast cancer in invasive breast cancer.

<p>Correlation of nuclear Kaiso with the molecular subtypes of breast cancer in invasive breast cancer.</p

Clinicopathological characteristics of 477 invasive breast cancer patients studied for the expression of Kaiso.

#<p>per 2 mm².</p>*<p>negative  =  N0 or N0(i+).</p>**<p>positive  =  ≥N1mi (according to TNM 7^th edition, 2010).</p

p120 and Kaiso localization in breast cancer cell lines.

<p>Human (A) and mouse (B) E-cadherin-expressing (top panels) and E-cadherin-deficient (bottom panels) breast cancer cell lines were stained for p120 (left panels) and Kaiso (middle panels). Right panels depict the merge of Kaiso (green) and p120 (red). Note the nuclear accumulation in MCF10a and Trp53^Δ/Δ-3 (arrows; also upper panels in C) <i>versus</i> cytoplasmic Kaiso expression in human and mouse ILC (arrowheads; IPH-926 and mILC-1). (C) Overexpression of p120 in Trp53^Δ/Δ-4 cells resulted in decreased nuclear accumulation of Kaiso (arrowheads; bottom panels) compared to untransfected Trp53^Δ/Δ-4, which shows predominantly nuclear Kaiso (arrows; upper panels). Size bars equal 20 µm. (D) Kaiso-dependent reporter assay using the 4XKBS reporter in mILC-1 and Trp53^Δ/Δ-4 cells. Upon overexpression of p120 in Trp53^Δ/Δ-4 cells, Kaiso-dependent gene repression is attenuated, whereas exogenous expression of Kaiso in mILC-1 increased gene repression.</p

E-cadherin and p120 membrane localization correlates with nuclear Kaiso expression.

<p>IDC (left panels) and ILC (right panels) were stained for E-cadherin, p120, and Kaiso using immunohistochemistry. Note the association between membrane-localized E-cadherin and p120, and high nuclear Kaiso in IDC. In contrast, ILC is characterized by loss of E-cadherin, and expression of cytoplasmic p120, which correlates with absence of nuclear Kaiso. Size bars equal 50 µm.</p

Correlation between functional adherens junctions and nuclear Kaiso expression in invasive breast cancer.

<p>Correlation between functional adherens junctions and nuclear Kaiso expression in invasive breast cancer.</p

Correlation of nuclear Kaiso with clinicopathological features in invasive breast cancer.

<p>Correlation of nuclear Kaiso with clinicopathological features in invasive breast cancer.</p