Avaliação molecular e imunohistoquímica da uretra de ratas após trauma induzido e terapia intravenosa com células-tronco derivadas de músculo

• Ano de defesa: 2017
• Autor(a) principal: Bilhar, Andreisa Paiva Monteiro [UNIFESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Stem cells; Urinary incontinence; Urethra; Incontinência urinária; Células ronco; Uretra
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5760899; http://repositorio.unifesp.br/handle/11600/49880

Objective: To identify the migration to the urethra of muscle derived stem cells (MDSCs) after intravenous (IV) injection in rats submitted to pelvic floor trauma due to vaginal distension (DV) and to analyze the effects of MDSC in the urethra of rats after seven and thirty days of the trauma in regards to: (1) mRNA expression of collagens, vascular endothelial growth factor (VEGF), nerve growth factor (NGF), Ki67 cell proliferation marker, and the expression of genes related to smooth and striated muscle apparatus; (2) expression of smooth and striated muscle proteins. Methods: MDSCs expressing green fluorescent protein (GFP) were injected into the tail vein of the rats at day three after DV. The urethras of five groups were analyzed: Control, Trauma 7 days (animals submitted to DV, sacrificed 7 days), MDSC 7 days (animals submitted to DV receiving IV therapy with MDSC, sacrificed 7 days), Trauma 30 days (animals submitted to DV, sacrificed at 30 days), MDSC 30 days (animals submitted to DV receiving IV therapy with MDSC, sacrificed at 30 days). Location of GFP cells was verified at 2 hours and 7 days after IV injection. RT-qPCR was used for gene expression and immunohistochemistry for protein identification and quantification. Results: MDSCs were early identified in the urethra, but only deformed cells after 7 days. After seven days of trauma, the MYH11 gene was overexpressed in the Trauma group in relation to Control (p=0.01). Expression of the mKi67 gene was increased in the MDSC group relative to Trauma (p=0.03) and Control (p=0.02). Expression of the COL1A1 and COL3A1 genes increased in the MDSC group relative to the Control (p=0.001 and p=0.002). The gene expression of NGF was decreased in the MDSC group in relation to Trauma (p=0.001) and there was no difference in expression of the MYH2, Desmin and VEGF gene. In the same period, immunohistochemical expression of MHY11, MYH2 and Desmin was increased in the MDSC group in relation to Trauma (p=0.01, p=0.005 and p=0.02) and decreased in the Trauma group in relation to the Control (p =0.01, p=0.008 and p=0.03). After 30 days of trauma, MYH11 and MYH2 gene expression was increased in the Trauma group compared to Control (p=0.03 and p=0.04). Expression of the mKi67 gene was increased in the MDSC group relative to Trauma (p=0.02) and Control (p=0.01). Expression of the COL1A1 gene increased in the MDSC group compared to the Control (p=0.008) and Trauma (p=0.03) and COL3A1 gene expression increased in the MDSC group compared to Control (p=0.03). However, the gene expression of NGF was decreased in the MDSC group in relation to Trauma (p=0.002) and there was no difference in the gene expression of Desmin and VEGF. In the same period, immunohistochemical expression of MYH11, MYH2 and Desmin was increased in the MDSC group in relation to Trauma (p=0.01, p=0.002 and p=0.01) and MYH2 gene expression decreased in the Trauma group compared to Control (p=0.04). Conclusion: MDSCs migrated early to the traumatized urethra, but did not integrate into the tissue. IV administration of MDSC alters the expression of genes related to cell proliferation, neural growth factor and extracellular matrix and the expression of smooth and striated muscle proteins in traumatized rat urethra.