2 research outputs found
Impact of galectin-1 gene deficiency on experimental diabetes mellitus induced by streptozotocin
A galectina-1 (Gal-1) é uma proteína que se liga à glicanas que possuem resíduos de β-galactosídeos e apresenta propriedades imunorreguladoras. Este estudo investigou a influência da Gal-1 endógena na doença autoimune diabetes mellitus tipo 1 induzida experimentalmente por estreptozotocina (STZ). Camundongos C57BL/6 selvagens (WT) e deficientes para o gene da galectina-1 (Gal-1 KO) foram tratados com STZ e monitorados quanto aos níveis sanguíneos de glicose. A análise histopatológica dos tecidos pancreáticos destes animais foi feita pela quantificação de células mononucleares infiltradas no interior das ilhotas de Langherans. Além disso, foi desenvolvido e validado o método analítico para determinação de alfa STZ no plasma dos camundongos utilizando cromatografia líquida de alta eficiência com detecção por espectrômetro de massas (CLAE-EM). Tal método apresentou faixa de linearidade de 0,312 a 160 μg/mL e ausência de efeito matriz. Os controles de qualidade nos ensaios de precisão e exatidão apresentaram coeficiente de variação e erro padrão relativo com valores inferiores à 15 %. Com relação à indução do diabetes, o tratamento de animais WT com STZ provocou consistente aumento da glicemia (382,88 mg/dL ± 72,31) bem como do infiltrado mononuclear nas ilhotas pancreáticas (169±48 células mononucleares/ilhota) em relação aos animais do grupo controle. Por outro lado, os níveis glicêmicos de camundongos Gal-1 KO tratados com STZ apresentaram menor proeminência, em relação aos animais WT, com valores médio 252,66 mg/dL ± 26,17. Padrão similar de atenuação também foi observado nas análises do infiltrado mononuclear nas ilhotas pancreáticas (94,7 ± 32,5 células mononucleares/ ilhota). Sendo assim, conclui-se que a ausência da Gal-1 endógena torna os camundongos resistentes à indução do diabetes mellitus tipo 1 por STZ, de modo independente à biodisponibilidade deste fármaco.Galectin-1 (Gal-1) is a protein that binds to glycans with β-galactoside residues and exhibits immunoregulatory properties. This study investigated the influence of endogenous Gal-1 on the experimental induction of type 1 diabetes mellitus, an autoimmune disease, by streptozotocin (STZ). C57BL/6 wild type (WT) and deficient mice for the galectin-1 gene (Gal-1 KO) were treated with STZ and monitored by determination of glycemia The histopathological analysis of the pancreatic tissues of these animals was performed by quantifying the mononuclear cells infiltrated inside the islets of Langherans. Furthermore, an analytical method for the determination of alpha-STZ in mice plasma was developed and validated using high-performance liquid chromatography with detection by mass spectrometer (HPLC-MS). The method presented linearity range of 0.312 to 160 μg/mL and also presented acceptable selectivity, accuracy and precision, as well as negligible matrix effect. WT mice treated with STZ showed a robust hyperglycemia (382.88 mg/dL ± 72.31), while this drug caused a discrete change in glycemic levels in Gal-1 KO mice (252.66 mg/dL ± 26.17). The number of mononuclear cells infiltrated in the islets of STZ-treated WT mice (169 ± 48 mononuclear cells/islet) was higher than the numbers of vehicle-treated WT mice (82 ± 22 mononuclear cells/islet) or Gal-1 KO treated mice with STZ (94.7 ± 32.5 mononuclear cells / islet). Finally, it is suggested that the absence of endogenous Gal-1 makes the Gal-1 KO mice resistant to the induction of type 1 diabetes mellitus by STZ, independently of the bioavailability of this drug
Induced Pluripotent Stem Cell for the Study and Treatment of Sickle Cell Anemia
Sickle cell anemia (SCA) is a monogenic disease of high mortality, affecting millions of people worldwide. There is no broad, effective, and safe definitive treatment for SCA, so the palliative treatments are the most used. The establishment of an in vitro model allows better understanding of how the disease occurs, besides allowing the development of more effective tests and treatments. In this context, iPSC technology is a powerful tool for basic research and disease modeling, and a promise for finding and screening more effective and safe drugs, besides the possibility of use in regenerative medicine. This work obtained a model for study and treatment of SCA using iPSC. Then, episomal vectors were used for reprogramming peripheral blood mononuclear cells to obtain integration-free iPSC. Cells were collected from patients treated with hydroxyurea and without treatment. The iPSCP Bscd lines were characterized for pluripotent and differentiation potential. The iPSC lines were differentiated into HSC, so that we obtained a dynamic and efficient protocol of CD34+CD45+ cells production. We offer a valuable tool for a better understanding of how SCA occurs, in addition to making possible the development of more effective drugs and treatments and providing better understanding of widely used treatments, such as hydroxyurea