RESUMO
Trichomonas vaginalis (Tv) induces host cell damage through cysteine proteinases (CPs) modulated by iron. An immunoproteomic analysis showed that trichomoniasis patient sera recognize various CPs, also some of them are present in vaginal washes (VWs). Thus, the goal of this work was to determine whether TvCP2 is expressed during infection and to assess the effect of iron on TvCP2 expression, localization and contribution to in vitro cellular damage. Western-blotting (WB) assays using TvCP2r and vaginitis patient serum samples showed that 6/9 Tv (+) but none of the Tv (-) patient sera recognized TvCP2r. WB using an anti-TvCP2r antibody and VWs from the same patients showed that in all of the Tv (+) but none of the Tv (-) VWs, the anti-TvCP2r antibody detected a 27 kDa protein band that corresponded to the mature TvCP2, which was confirmed by mass spectrometry analysis. Iron decreased the amount of TvCP2 mRNA and the protein localized on the parasite surface and cytoplasmic vesicles concomitant with the cytotoxic effect of TvCP2 on HeLa cells. Parasites pretreated with the anti-TvCP2r antibody also showed reduced levels of cytotoxicity and apoptosis induction in HeLa cell monolayers. In conclusion, these results show that TvCP2 is expressed during trichomonal infection and plays an important role in the in vitro HeLa cell cytotoxic damage under iron-restricted conditions.
Assuntos
Cisteína Proteases/metabolismo , Ferro/administração & dosagem , Proteínas de Protozoários/metabolismo , Trichomonas vaginalis/efeitos dos fármacos , Vagina/parasitologia , Secreções Corporais/parasitologia , Feminino , Humanos , Trichomonas vaginalis/enzimologiaRESUMO
OBJECTIVES: To determine the effects of W100E-Leptin in a streptozotocin-induced diabetic mice model (effects in the body weight, fasting serum glucose and glucose tolerance). METHODS: Intraperitoneal W100E-Leptin application at 1 mg/kg for 13 days. We used 3 experimental groups (n=6). Group 1: Diabetes + W100E-Leptin (intraperitoneal administration), Group 2: Diabetes + buffer (vehicle) and Group 3: Healthy control + buffer (vehicle). RESULTS: We determined the effects of W100E on the behavior of the mice, more active, more hair and a tendency to gain body weight. We did not observe any hypoglycemic effect of W100E-Leptin on serum glucose levels in the tests we performed. CONCLUSIONS: These results show us the need to characterize the effects of this hormone in diabetes. We will continue with the characterization of the change that is generated in the protein regulation caused by W100E-Leptin in the diabetes, to propose this hormone as an adjunct against diabetes
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