Papel do ferro e da Heme-oxigenase-1 na toxoplasmose e malária congênitas
Publicado: 07/10/2018 - 11:17
Última modificação: 07/10/2018 - 11:17
In congenital toxoplasmosis when primary infection is acquired in the first trimester, the parasite can be transmitted to fetus and cause inflammatory lesion that may lead to neurological damage and abortion. The risk of abortion is high in women infected in the third trimester and low in the first one. In contrast, the risk of abortion is higher in those infected in the first trimester. In Plasmodium infection during pregnancy it could be serious complications, such as spontaneous abortion, intrauterine growth retardation, and low birth weight, which are known risk factors for neonatal mortality. In the present study we intend to verify the role of iron in congenital toxoplasmosis in mouse experimental model and in vitro in BeWo trophoblastic human cells. We also intend to verify the role of iron in congenital malaria. For this purpose, BeWo cells will cultured and treated with IFN-g or IL-6 or iron-chelating agent deferoxamine or iron supplementation with ferric ammonium citrate (FAC), inducer of Heme-oxygenase-1 (CoPP), and cells will be infected with 2F1 strain of Toxoplasma. For Toxoplasma in vivo experiments, C57BL6 females will be housed with male and on the day of vaginal plug detection (first day of gestation) will treated with ferric ammonium citrate every day or with deferoxamine or CoPP on days 3, 5 and 7 post-infection. In parallel, groups of C57BL6 mice will house at the same way, infected with 103 red blood cells systemically, and treated as described above. The mortality and absorption rate will be recorded every day. For Plasmodium infection the parasitemia will be analyzed by flow citometry and parasitism in the placenta by histology. For Toxoplasma quantification the tissue parasitism in the placenta/uterus will be assayed by immunohistochemistry and qPCR. The expression of HO-1, hepcidin, transferrin receptor, lactoferrin, BcLXl and DMT1 in placenta/uterus from mice infected and treated or in BeWo infected and treated cells will be analyzed by qPCR with human or mouse specific primers. To verify the apoptotic index, the expression of HO-1 and BclXL, caspase-3 cleaved and non-cleaved will be verified in BeWo or placenta/uterus tissues by immunoblotting or immunohistochemistry. It was also verified by immunohistochemistry using TUNEL kit. The cytokine profile will verified by flow citometry with citometric beads array (CBA). The results obtained with these experiments will bring new knowledge and strategies of treatment for congenital toxoplasmosis and malaria.